The purpose of the present investigation was to explore the effects of well-defined flow conditions on the activity of tissue factor (TF) expressed on the surface of cultured rat vascular smooth muscle cells. Cells were cultured to confluence on Permanox brand slides and stimulated to express TF by a 90 min incubation with fresh growth medium containing 10 percent calf serum. The stimulated cells were then placed in a parallel plate flow chamber and perfused with Hanks Balanced Salt Solution containing factor VIIa, factor X (FX), and calcium. The chamber effluent was collected and assayed for factor Xa (FXa) and the steady-state flux of FXa was calculated. The flux values were 68.73, 94.81, 139.75, 138.19, 316.82, and 592.92 fmole/min/cm2 at wall shear rates of 10, 20, 40, 80, 320, and 1280 s−1 respectively. The FXa flux depended on the wall shear rate to a greater degree than predicted by classical mass transport theory. The flux at each shear rate was three to five times less than that ...

TY - JOUR. T1 - Modulation of collagen synthesis by tumor necrosis factor alpha in cultured vascular smooth muscle cells. AU - Hiraga, Syouichi. AU - Kaji, Toshiyuki. AU - Ueda, Yoshimichi. AU - Zisaki, Fumiko. AU - Iwata, Kazushi. AU - Koizumi, Fumitomo. AU - Okada, Yasunori. AU - Katsuda, Shogo. AU - Nakanishi, Isao. PY - 1999/12/10. Y1 - 1999/12/10. N2 - Collagen synthesis in vascular smooth muscle cells (SMCs) after exposure to tumor necrosis factor alpha (TNF-α) was investigated using a culture system. The synthesis of collagenase-digestible proteins (CDP) and noncollagenous proteins (NCP) was evaluated by the [3H]proline incorporation. It was shown that TNF-α markedly suppresses the incorporation of [3H]proline into both CDP and NCP in confluent cultures of SMCs but not in sparse cultures of the cells. Such a marked suppression by TNF-α was not observed in confluent bovine aortic endothelial cells and human fibroblastic IMR-90 cells. In confluent SMCs, the synthesis of CDP was more ...

I am planning to set up a co-culture system for human endothelial cells and human VASCULAR smooth muscle cells. I would be really interested to find out about methods of extraction and primary culture of human vascular smooth muscle cells, if anyone can help and advise, please mail me! Thanks Pippa Deex ...

Vascular smooth muscle (VSM) cell proliferation contributes to the pathogenesis of atherosclerosis, restenosis after angioplasty and vein graft disease. The regulation of genes involved in VSM cell proliferation, particularly by naturally occurring inhibitors, is therefore of some importance. We have investigated the role of the c-myc proto-oncogene in growth arrest of exponentially proliferating rat VSM cells, following mitogen withdrawal, treatment with heparin (50 micrograms/ml), interferon-gamma (IFN-gamma) (100 i.u./ml), or the cyclic nucleotide analogues, 8-bromo-adenosine-3′5′-cyclic monophosphate (8-Br-cAMP; 0.1 mM) and 8-bromoguanosine-3′5′-cyclic monophosphate (8-Br-cGMP; 0.1 mM). Growth arrest was accompanied by down-regulation of c-Myc protein and mRNA following treatment with all inhibitors. Serum withdrawal or IFN-gamma treatment suppressed c-myc expression by more than 50% within 2 h, and this occurred throughout the cell cycle. Platelet-derived growth factor, epidermal ...

The concept of arterial SMC heterogeneity has gained wide acceptance in the last years.1 2 33 The distinct phenotypes of arterial SMCs have been mainly identified in vitro,4 5 6 7 8 9 10 11 12 13 14 15 16 suggesting that specific features of SMC populations arise and are maintained in the particular environment of cell culture. Hence, it was of interest to investigate whether in vitro SMC phenotypes are preserved when SMCs are placed back in an in vivo environment. For this purpose, we have implanted 2 SMC populations exhibiting distinct levels of differentiation in vitro into the rat carotid artery submitted to endothelial injury.24 25 The implanted SMCs were marked with PKH-26, a lipophilic cell membrane linker that is halved with each cell division but is not lost from the cell membrane.34 Our results show that the 2 implanted populations essentially retain for 20 days in vivo the phenotype that they specifically exhibited in vitro.. Spindle-shaped and epithelioid rat SMC populations have ...

Primary monolayer cultures were obtained in 60-mm petri dishes by incubating 3 X 10(6) isolated hepatocytes at 37 degrees C in Dulbeccos medium supplemented wi

Thank you for your interest in spreading the word about Biochemical Journal.. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.. ...

Jochen Meier Ivonne Strmel Radu Iosub Sonja Schmidt and Rosemarie Grantyn Developmental Physiology Johannes Mller Institute Humboldt University Medical School (Charit) Berlin Germany Efficient delivery of DNA to primary neuronal cell cultures is of critical importance,Effectene,Transfection,Reagent,provides,efficient,gene,delivery,,,,,,,,,,,,to,primary,neuronal,cell,cultures,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology

Jacobs, B B., In vivo assay of function of mouse ovaries following culture in hormone enriched medium. (1963). Subject Strain Bibliography 1963. 546 ...

Full Text - Hydrogen peroxide (H2O2) induces oxidative injury to human osteoblasts. The expression and potential function of circular RNA HIPK3 (circHIPK3) in H2O2-treated human osteoblasts were tested. We show that H2O2 significantly downregulated circHIPK3 in OB-6 cells and primary human osteoblasts. Furthermore, circHIPK3 levels were decreased in the necrotic femoral head tissues of dexamethasone-treated patients. In OB-6 osteoblastic cells and primary human osteoblasts, forced overexpression of circHIPK3 by a lentiviral construct alleviated H2O2-induced viability reduction, cell death and apoptosis. Contrarily, circHIPK3 silencing by targeted shRNA potentiated H2O2-induced cytotoxicity in OB-6 cells and primary human osteoblasts. Moreover, circHIPK3 downregulation by H2O2 induced miR-124 accumulation in OB-6 cells and primary human osteoblasts. On the contrary, miR-124

Many studies have shown that TRAIL is a potent apoptosis inducer in malignant cells, whereas its role in normal cell physiology is much less well understood.1 We have demonstrated here that both aortic endothelial cells and HUVECs exhibit a similar pattern of surface TRAIL-R expression. Moreover, in vascular endothelial cells, TRAIL stimulates the phosphorylation of the serine/threonine kinase Akt in a manner dependent on PI3K activation.14 The ability of TRAIL to activate the antiapoptotic PI3K/Akt pathway in endothelial cells is a completely new and unexpected finding, also in consideration of the large number of studies underlining the proapoptotic activity of TRAIL, at least in malignant cells.1,4,5 It should be underlined that the PI3K/Akt pathway is of central importance in endothelial cell biology, conferring survival to endothelial cells in response to angiogenic cytokine stimulation, fluid shear stress, and matrix attachment signals. This pathway is essential also for endothelial cell ...

TY - JOUR. T1 - Activation of caspase-8 and Erk-1/2 in domes regulates cell death induced by confluence in MDCK cells. AU - Chang, Yung Heng. AU - Lin, Hsi Hui. AU - Wang, Yang Kao. AU - Chiu, Wen Tai. AU - Su, Hsiao Wen. AU - Tang, Ming Jer. PY - 2007/4/1. Y1 - 2007/4/1. N2 - Under normal culture conditions, cells adhere to culture dish, spread out, proliferate, and finally cover all areas and reach confluence. During the confluent stage, cell proliferation ceases and differentiation is enhanced. Meanwhile, cell death also appears as the monolayer confluence proceeds. To delineate the mechanism of cell death induced by the confluent process, we employed Madin-Darby canine kidney (MDCK) cells. When approaching confluence, MDCK cells exhibited increase the levels of caspase-2 and enhanced the activity of caspase-8. Using various caspase inhibitors to block apoptosis, we found that only z-VAD-fmk and z-IETD-fmk can inhibit confluent cell death, indicating that confluent cell death is mediated by ...

In the present article, we compared the antiangiogenic activity of trans- and cis-resveratrol on endothelial cells of different origin activated by endogenous or exogenous stimuli. The results indicate that trans-resveratrol affects endothelial cell proliferation and various cell proliferation-independent aspects of the angiogenesis process, including endothelial cell sprout formation, collagen gel invasion, and morphogenesis. Thus, trans-resveratrol acts as an angiostatic molecule on multiple targets of the angiogenic process. In all the assays, the cis stereoisomer exerted only a limited effect, if any. In keeping with the in vitro observations, trans-resveratrol was more effective than the cis isomer in inhibiting angiogenesis and tumor growth in vivo.. The molecular bases of the antiangiogenic activity of resveratrol are poorly understood (16). trans-Resveratrol is endowed with a significant antioxidant and anti-inflammatory activity (8, 9). However, no direct relationship exists between the ...

TY - JOUR. T1 - Recombinant murine and human IL 1α bind to human endothelial cells with an equal affinity, but have an unequal ability to induce endothelial cell adherence of lymphocytes. AU - Thieme, T. R.. AU - Hefeneider, S. H.. AU - Wagner, C. R.. AU - Burger, D. R.. PY - 1987/1/1. Y1 - 1987/1/1. N2 - Consistent with the reports of others, we have demonstrated that human peripheral blood lymphocytes adhere to cultured human umbilical vein-derived endothelial cells (EC) in vitro. In our studies adherence was increased twofold to threefold by a 6-hr preincubation of the EC with IL 1. Recombinant human IL 1α induced a maximal adherence response at less than 1 U per 2 x 104 EC. In contrast, recombinant murine IL 1α was found to be 250- to 1250-fold less active in the adherence assay, based on units of IL 1 activity defined by the murine thymocyte proliferation assay. Moreover, when EC were preincubated with excess murine IL 1, no inhibition of the adherence-inducing effect of human IL 1 was ...

The effects of interferons (IFNs) on lymphocyte adhesion to cultured human vascular endothelial cells (EC) were investigated using an in vitro assay. Endothelial cells obtained from umbilical vein were first cultured at a low density with a conditioned medium (CM) from 12-O-tetra decanoylphorbol 13-acetate-concanavalin A (TPA-Con A) stimulated human peripheral blood lymphocytes (PBL), or with recombinant (r) gamma interferon (IFN-gamma) or r alpha interferon (IFN-alpha), and then were incubated with freshly isolated PBL. Natural IFN-gamma in the TPA-Con A CM and rIFN-gamma (12.5-500 U/ml) induced major histocompatibility complex-class II antigens (HLA-DR, HLA-DP, and HLA-DQ) and significant lymphocyte adhesion to the EC, whereas rIFN-alpha did not. The lymphocyte adhesion to the EC and the expression of DR antigens on the EC were well correlated in terms of both kinetics and the dose-response pattern of rIFN-gamma. When EC expressing I region associated (Ia) antigen were preincubated with ...

Human Pulmonary Fibroblast Cell Pellet https://www.sciencepro.com.br/produtos/sc-cp3300 https://www.sciencepro.com.br/@@site-logo/logo-novo.png ...

The fully sequenced pCERI is transformed into E.coli BL21 and E.coli Nissle 1917 that are used for the characterization of our regulatory system. The expression efficiency of the genes of interest is characterized by a fluorescence-based assay (Results) Overnight cultures of both recombinant strains are inoculated with single colonies. The overnight cultures are diluted with LB-Amp to an OD of 0.02. 200 μ of the dilutions are pipetted in microtiter plates and incubated at 37 °C in the plate reader. Every 15 minutes mRFP fluorescence (Ex. 584 nm, Em. 607 nm), CFP fluorescence (Ex. 439 nm, Em. 476) and OD600 is measured over a period of 13 h. The measured RFU (relative fluorescence unit) is either blotted against time or against OD600 to evaluate the expression behavior of pCERI. Additionally several samples are either induced with 0.1 -50 nM of 3OC6-HSL and 0.1-100 nM of C8-HSL to explore putative advantages of higher HSL-synthesis, that might be possible by positioning each HSL-synthase, ...

This unique comprehensive volume in the Human Cell Culture Series encompasses research methodology for the growth and differentiation of all types of primary mesenchymal cells. Each of eleven mesenchymal cell types is covered by a leading expert in the field, providing insightful background information along with detailed current culture and assay techniques. Tissue types covered include bone, cartilage, tendons and ligaments, muscle (smooth, skeletal, and cardiac), adipose tissue, and skin (dermis), as well as multi-potent mesenchymal stem cells and fibrocytes. In addition to uses for research applications, current and future clinical applications of large-scale culture methods are also discussed. Further, the procurement and processing of primary human tissues is covered in detail. The generous inclusion of figures and tables, along with a consistent format, makes critical information highly accessible to the reader. Additionally, numerous literature citations provide a valuable reference for ...

Choice of AMP 100 versus AMP 50 - posted in Molecular Cloning: Ok, i know the main difference is the amp concentration. But i wonder if it matters that i use amp 50 for overnight culture versus amp 100 for overnight culture.

I am studying the differentiation of leukemic cells along the monocytic lineage using calcitriol as differentiative agent and others vitamins as possible stimulating compounds on differentiation. In my work I have used HL-60 cells as in vitro culture cells but now I have some problems because these cells have changed their response and develop expontaneous differentiation in normal culture conditions. Now, I am thinking on NB4 cells as a better cell line to study differentiation. I have ask some information to cell banks as ATCC and ECA but they dont have this cell line. Could anybody tell me if anyone can send me this cell line?, I have very gratefull if anyone could send me HL-60 cells too. Waiting positive responses and acknowledging in advance. Guillermo Lopez-Lluch Departamento de Biología Celular Facultad de Ciencias Universidad de Córdoba Avda. S. Alberto Magno S/N E-14007 Córdoba, Spain. TFNO: Int + 34-57-218634 FAX: Int + 34-57-218634 e-mail: bc2biced at lucano.uco.es ...

TY - JOUR. T1 - Cell interactions in nerve and muscle cell cultures.. AU - Christian, C. N.. AU - Bergey, G. K.. AU - Daniels, M. P.. AU - Nelson, P. G.. PY - 1980/12/1. Y1 - 1980/12/1. N2 - The neurotransmitter synthesized by a given class of neurones is subject to modification and, indeed, a qualitative switchover in transmitter biochemistry recently has been demonstrated (Furshpan, POtter & Landis, 1980; Walicke, Campenot & Patterson, 1977). In conjunction with the specification of transmitter biosynthesis that becomes established in a given neurone, a complementary specification of appropriate receptor production is required in any cell functionally post-synaptic to that neurone. An additional requirement of peculiar force in the nervous system has to do with the spatial organization of the receptors in the surface membrane of the post-synaptic cell once the receptors are synthesized. Inappropriately distributed receptors are useless receptors. The perfect registration of a variety of types ...

I have provided this website and/or the information for educational and community purposes only. Nothing contained in here is intended to be instructional for medical diagnosis or treatment. Although I make a conscientious effort to provide high quality information, I disclaim any implied guarantee about the accuracy, completeness, timeliness or relevance of any information. For any questions concerning medical needs, visit or call your CF Center or health care professional. This web site is meant for the community for support and information. I do not give medical instruction or treatment. All health information found on this blog including search engine resources, may not be right for your medical conditions and needs. Always consult your doctor, healthcare specialist/practitioner or medical center when determining which course of health care regime is best for your individual care.. ...

TY - JOUR. T1 - Studies on the mechanism of hrmonal stimulation of zinc uptake in human cell cultures. T2 - hormone-cell interactions and characteristics of zinc accumulation.. AU - Cox, R. P.. AU - Ruckenstein, A.. PY - 1971/2. Y1 - 1971/2. UR - http://www.scopus.com/inward/record.url?scp=0015012917&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=0015012917&partnerID=8YFLogxK. M3 - Article. C2 - 4322943. AN - SCOPUS:0015012917. VL - 77. SP - 71. EP - 81. JO - Journal of Cellular Physiology. JF - Journal of Cellular Physiology. SN - 0021-9541. IS - 1. ER - ...

One group of projects in the lab concerns co-receptors, cell-surface molecules that regulate the interaction between soluble growth factors and their cell-surface receptors. We have identified a novel co-receptor for BMPs (bone morphogenetic proteins) and are studying the roles of Heparan Sulfate Proteoglycans (HSPGs) as co-receptors for a variety of growth factors, including those that drive tumor development and pattern the developing nervous system. We have used knockout technology to generate mice that are mutant for particular HSPGs, and are also working with mice that are mutant for some of the biosynthetic enzymes required to make HSPGs. We are also using modeling approaches to understand how HSPGs influences the kinetic parameters of growth factor-receptor interaction ...

Were stained with rat endothelial cell antigen. + pixels were counted in glomeruli and tubular fields working with ImageJ Application . To be able to evaluate

How would the cells respond to Aβ? Mancuso injected 5 μL of 10 μM synthetic oligomers into the mouse brain ventricles eight to 10 weeks after transplanting the human microglia. In reaction to this insult, endogenous mouse microglia shifted their transcriptomes sequentially from homeostatic to cytokine responsive, to activated, with the latter partially overlapping with disease-associated microglial (DAM) signatures described previously (Jun 2017 news). Human microglia in the mice underwent a transcriptional transformation as well, but analysis of more than 10,000 orthologous genes indicated poor correlation between human and mouse microglial responses. Of 207 differentially activated genes, 112 were up in human but not mouse microglia; they included GWAS hits BIN1 and PICALM. We saw that human microglia responded very differently to Aβ oligomers. This emphasizes the need to look specifically at human cells in the context of AD, said De Strooper.. Blurton-Jones and colleagues took a ...

https://doi.org/10.18632/oncotarget.26640 Amruta Phatak, Mohammad Athar, James A. Crowell, David Leffel, Brittney-Shea Herbert, Allen E. Bale, Levy Kopelovich

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Recent evidence supports a strong role for immune suppression in facilitating cancer progression. Transforming growth factor beta (TGFß) regulates a diverse array of biological events, including neoangiogenesis, stromal formation, and immune responses. Both TGFß and a small molecule inhibitor of TGFßR-1, LY364947, were effective regulators of these processes in vitro. Using quantitative RT-PCR and fluorescence imaging, TGFß increased collagen production by pancreatic stellate cells while addition of LY364947 suppressed production below basal levels. Endothelial cell proliferation, measured as a function of endothelial tube formation in Geltrex® matrix, was similarly increased by TGFß and reduced to basal levels by LY364947. To explore the in vivo effects of these molecules, LY364947 was encapsulated in liposomes to encourage tumor accumulation and to avoid administration of DMSO vehicle. Liposomes were prepared with a molar ratio of 6:3:1 1,2-dipalmitoyl-sn-glycero-3-phosphocholine ...

Simple muscle cell cultures are utilized frequently to investigate the mobile mechanisms of contraction. in Personal computer and RC vs .. Me personally and FC (all < 0.05). Appearance of Gi3, serine/threonine proteins phosphatase-1 -catalytic subunit, and Rho kinase 1 improved in Personal computer and RC vs .. Me personally and FC (all < 0.05). Cell tradition and resuspension downregulated appearance of -actin and calponin, but not really myosin weighty string. The online impact of these molecular adjustments was reductions of cell reactivity to ACh in RC vs .. FC. Overexpression of CPI-17 in Personal computer partly reversed the reductions of contractility in resuspended cells. Methylation-specific PCR demonstrated improved methylation of the gene marketer in Computer vs .. Me personally (< 0.05). We agreed that even muscles cells preserve their contractile phenotype in lifestyle. Nevertheless, reactivity to ACh diminishes because of changed reflection of particular cell-signaling protein ...

Conventional biology expects the process of mammalian cell division, mitosis, to occur by the equal partition of a mother cell into two daughter cells. Bioengineers at UCLA Engineering have developed a platform that mechanically ...

A growing body of evidence suggests that transiently activated NF-κB in noninflammatory states and persistently activated NF-κB during inflammation play different pathophysiological roles in vivo. During the initial phase of inflammation, proinflammatory cytokines and mediators induce prolonged NF-κB activation in various inflammatory cells and endothelium. The activated NF-κB further upregulates the expression of several proinflammatory molecules, such as endothelial cell adhesion molecules and macrophage inflammatory protein-2, which triggers neutrophilic infiltration and tissue injury (1, 27, 34). Thus persistent NF-κB activation during the early phase of inflammation amplifies inflammatory response in vivo.. In contrast, transient activation of NF-κB before inflammatory stimulation results in the anti-inflammatory response. For example, several investigators (41, 54, 56) have found that transient activation of NF-κB is required for the heart to tolerate ischemia-reperfusion-induced ...

Time-dependent induction of connected cells by overexpression of cdc5ΔN. (A) Strain KLY1083 expressing three copies of GAL1-EGFP-cdc5ΔN homogeneously induced

floating THP-1 cells - medium change advice - posted in Tissue and Cell Culture: Hello, I was adamantly asked to culture THP-1 cells and did so, by thawing a cryovial and seeding the cells in a 25 cm flask. 3 days later 99% of cells are floating, which I reckon is OK. Isnt it? So, do I need to change the medium before they reach ~8x 10^5 cells/ml and are ready to be passed? Since these are floating cells, how do I do that? Also, would a 1/3 split ratio be OK for these cells? Th...

1) The main issue with this paper is this: these cell lines, although originally human, are all immortalized cancer cell lines characterised by markedly different biological properties when compared to normal human cells of the same tissue/cell type. They cant be readily used a surrogates for normal human tissue/cell types. None were primary cells nor were any even from recently acquired tissue samples from biopsies etc. People have infected primary human airway epithelial cultures with hCoV-EMC - so this can be done successfully - , although it would be more difficult for other tissue types as these cultures havent been developed. Some of these cell lines used may by chance lack key viral repressors of infection present in normal primary cells, which could skew results from cell culture infection experiments. Plus, a human tissue is not just a single cell type - they are composed of diverse kinds of cells that could together behave much, much differently than cell lines in culture ...

Accepted name: raucaffricine β-glucosidase. Reaction: raucaffricine + H2O = D-glucose + vomilenine. For diagram click here.. Other name(s): raucaffricine β-D-glucosidase; raucaffricine glucosidase. Systematic name: raucaffricine β-D-glucohydrolase. Comments: Highly specific; some other ajmalan glucoside alkaloids are hydrolysed, but more slowly.. Links to other databases: BRENDA, EXPASY, KEGG, Metacyc, PDB, CAS registry number: 102925-37-1. References:. 1. Schübel, H., Stöckigt, J., Feicht, R. and Simon, H. Partial-purification and characterization of raucaffricine β-D-glucosidase from plant cell-suspension cultures of Rauwolfia serpentina benth. Helv. Chim. Acta 69 (1986) 538-547.. ...

Increasing numbers of studies have demonstrated that PGE1 plays an important role in determining the survival of cells exposed to H/R conditions. PGE1 treatment of liver sinusoidal endothelial cells increases their survival and resistance to apoptosis during H/R injury by suppressing NO production and MMP release [31]. Interestingly, a previous study by Ma et al. [32] reported that the total number of cardiomyocytes was decreased in an H/R group when compared with a group of cardiomyocytes pretreated with PGE1, and the mechanism for that effect probably involved a decrease in Bax expression and an increase in Bcl-2 expression. Caspase 3, which is downstream of Bcl-2 and Bax, is considered to be a critical protease required for apoptosis [39]. In the present study, we observed that inhibition of apoptosis by PGE1 in H/R injured cells involved the inactivation of cleaved-caspase-3. Our results support a model in which PGE1 inhibits apoptosis in cardiomyocytes, at least in part, via a ...

Finally, our article on human and mouse neurons has been published! The picture above shows the neurons of a human and a mouse. Human and mouse neurons are similar overall, but they also have…

CD7, also known as gp40, is a member of the immunoglobulin superfamily found on T cells, NK cells, thymocytes, hematopoietic progenitors, and monocytes (weakly). CD7 is also expressed on acute lymphocytic leukemia (ALL). CD7 crosslinking induces a calcium flux in T lymphocytes, presumably as a result of cytoplasmic domain association with PI3-kinase. CD7 co-stimulation can induce cytokine secretion and modulate cellular adhesion. A ligand of CD7, epithelial cell secreted protein K12, is produced in thymus to regulate thymocyte signaling and cytokine release. In lung microvascular endothelial cells CD7 serves as an IgM Fc receptor. Expression of CD7 is an important marker used in leukemia diagnostics ...

483279889 - EP 3108256 A4 2017-10-04 - METHOD OF DETECTING CANCER - [origin: WO2015120523A1] The disclosure provides methods for resolving an inconclusive cytological assessment of clinically relevant cells in a sample obtained from a patient based on the cytological detection of antibody binding to telomerase in clinically relevant cells in the sample, wherein binding of the antibody to clinically relevant cells indicates the presence of malignant or cancerous cells.[origin: WO2015120523A1] The disclosure provides methods for resolving an inconclusive cytological assessment of clinically relevant cells in a sample obtained from a patient based on the cytological detection of antibody binding to telomerase in clinically relevant cells in the sample, wherein binding of the antibody to clinically relevant cells indicates the presence of malignant or cancerous cells.

Results ICAM-1 expression was significantly up-regulated upon the introduction of TNF-α under all conditions in HUVEC (figure 1). However, baseline expression was increased when co-cultured with both NHDF (2.0 vs 1.3, p,0.001) and HUASMC (6.5 vs 1.3, p,0.001). This meant that ICAM-1 expression at 12 hours was also significantly higher in co-culture with NHDF (8.3 vs 5.2, p,0.001) and HUASMC (11.0 vs 5.2, p,0.001). Moreover, there was a moderate relationship between HUVEC ICAM-1 expression and the cell ratio when in co-culture with NHDF, where decreasing NHDF resulted in decreased ICAM-1 in HUVEC (R2=0.45).. TNF-α caused an increase in ICAM-1 expression in NHDF under monoculture conditions (Fig. 2); this up-regulation was significantly reduced in co-culture conditions with HUVEC (1.7 vs 5.1, p,0.001). A similar trend was observed when in co-culture with HUASMC (2.7 vs 5.1, p,0.001), except the baseline expression of ICAM-1 was also increased (2.8 vs 1.1, p,0.001).. Constitutive production of ...

Mouse IL-4 ELISA kit (5 plate). The U-CyTech sandwich ELISA kits have been developed for cytokine determinations in culture supernatant, serum and plasma. The kit is supplied with all necessary reagents to perform 480 ELISA determinations. Cytokines are a group of regulatory proteins critically involved in many physiological processes such as immune recognition, cell differentiation and cell proliferation. They have been identified in many vertebrate species and are produced by a variety of different cell types. Cytokines are usually produced transiently and locally, acting in a paracrine or autocrine manner. They interact with high affinity cell surface receptors specific for each cytokine or cytokine group and are active at very low concentrations mostly in the picogram range. It is well known now that the type of an antigen-specific immune response largely depends on the selection or preferential activation of defined CDM|sub|4|sup|+ T cell subsets (i.e. Th1 and Th2). Activation of these subsets is

Mouse IL-10 ELISA kit (5 plate). The U-CyTech sandwich ELISA kits have been developed for cytokine determinations in culture supernatant, serum and plasma. The kit is supplied with all necessary reagents to perform 480 ELISA determinations. Cytokines are a group of regulatory proteins critically involved in many physiological processes such as immune recognition, cell differentiation and cell proliferation. They have been identified in many vertebrate species and are produced by a variety of different cell types. Cytokines are usually produced transiently and locally, acting in a paracrine or autocrine manner. They interact with high affinity cell surface receptors specific for each cytokine or cytokine group and are active at very low concentrations mostly in the picogram range. It is well known now that the type of an antigen-specific immune response largely depends on the selection or preferential activation of defined CDM|sub|4|sup|+ T cell subsets (i.e. Th1 and Th2). Activation of these subsets is

2010) (Fig. 1A). Fibroblasts were seeded at 1.5 × 105 cells/filter and HUVEC were seeded at 1.0 × 105 cells/filter to yield confluent monolayers within 24 h. After 24 h, culture media were removed and the 24-well inserts were fitted into the 12-well inserts, with 200 μl fibroblast medium added to the surface of each filter and 1.5 ml to the lower chamber. Cells were co-cultured together for 48 h, with 100 U/ml TNF alpha (R&D Systems, Abingdon, UK) in combination with 10 ng/ml IFN gamma (Peprotech Inc., London, UK) added for the second 24 h when desired. For. comparison, parallel cultures of HUVEC or fibroblasts were maintained alone on their Selleckchem AC220 original filters. To form collagen gels, ice-cold rat-tail type 1 collagen PD-166866 cell line dissolved in acetic acid (2.15 mg/ml; First Link Ltd, West Midlands, UK) was mixed with ice cold 10 × concentrated M199 in the ratio 830:170 and the pH was neutralised by addition of ice cold 1 N NaOH. For each 1 ml of gel, 160 μl FCS was ...

You know its funny that you mention tube like structures...... do they look fairly long and rectangular? I have something growing in tissue culture that looks like that. These cells started off as hemangiomas. We used collangenase to break it up, and are growing these cells in DMEM with added penicillin/streptomyacin and fetal bovine serum. Ive seen quite a few of these, one person thinks they look like myocytes....heart muscle cells in particular, they arent beating or anythying though. And if that is what they are, id be really really surprised. Most of the cells look like fibroblasts (Im thinking that what look like fibroblasts might be pericytes) with some scattered possible endothelial cells. We transfected a few dishes with a vector to induce division, but we dont see these cells in any of the tissue culture dishes containing the transformed cells. I wonder if we are looking at the same thing. Ive ordered a camera for our inverted scope, ill have to take a picture when it comes in. ...

The characterization was performed using overnight cultures of the corresponding part. The optical density of the cell cultures was adjusted to 0.4 and 50 µl of the culture sample was pipette onto a microscope slide and a coverslip was placed over it. A 100X/1.3 Oil immersion objective was used for the light microscopy imaging and filters necessary for CFP fluorescence detection was set. An initial live imaging was done in transmitted light mode to find a field of view with maximum number of cells. The same was then used for capturing the fluorescence signal of the CFP. The same procedure was repeated several times in order to quantify the data. If we need to observe more number of cells under a given field of view, it is recommended to spin down the cell culture and use the resuspended pellet for increased cell density.. ...

The characterization was performed using overnight cultures of the corresponding part. The optical density of the cell cultures was adjusted to 0.4 and 50 µl of the culture sample was pipetted onto a microscope slide and a coverslip was placed over it. A 100X/1.3 Oil immersion objective was used for the light microscopy imaging and filters necessary for CFP fluorescence detection was set. An initial live imaging was done in transmitted light mode to find a field of view with maximum number of cells. The same was then used for capturing the fluorescence signal of the CFP. The same procedure was repeated several times in order to quantify the data. If we need to observe more number of cells under a given field of view, it is recommended to spin down the cell culture and use the resuspended pellet for increased cell density.. ...

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Fomin, Victor P., Blair E. Cox, and R. Ann Word. Effect of progesterone on intracellular Ca21 homeostasis in human myometrial smooth muscle cells. Am. J. Physiol. 276 (Cell Physiol. 45): C379-C385, 1999.-Although it is well known that progesterone alters uterine contractility and plays an important role in maintenance of pregnancy, the biochemical mechanisms by which progesterone alters uterine contractility in human gestation are less clear. In this investigation we sought to identify progesterone-induced adaptations in human myometrial smooth muscle cells that may alter Ca21 signaling in response to contractile agents. Cells were treated with vehicle or the progesterone analog medroxyprogesterone acetate (MPA) for 5 days, and intracellular free Ca21 concentration ([Ca]i) was quantified after treatment with oxytocin (OX) or endothelin (ET)-1. OXand ET-1-induced increases in [Ca]i were significantly attenuated in cells pretreated with MPA in a dose-dependent manner. Progesterone receptor antagonists

TY - JOUR. T1 - The mechanisms of uremic serum-induced expression of bone matrix proteins in bovine vascular smooth muscle cells. AU - Chen, N. X.. AU - Duan, D.. AU - ONeill, K. D.. AU - Wolisi, G. O.. AU - Koczman, J. J.. AU - LaClair, R.. AU - Moe, S. M.. PY - 2006/9/1. Y1 - 2006/9/1. N2 - We have previously found that uremic human serum upregulates RUNX2 in vascular smooth muscle cells (VSMCs), and that RUNX2 is upregulated in areas of vascular calcification in vivo. To confirm the role of RUNX2, we transiently transfected a dominant-negative RUNX2 (ΔRUNX2) construct in bovine vascular smooth muscle cells (BVSMCs). Blocking RUNX2 transcriptional activity significantly decreased uremic serum induced alkaline phosphatase (ALP) activity (268 ± 34 vs 188 ± 9.5 U/g protein, P , 0.05) and osteocalcin expression (172 ± 17 vs 125 ± 9 ODU, P , 0.05). To determine the mechanism by which uremic serum upregulates RUNX2, we examined cell signaling pathways. BVSMCs were incubated in the presence or ...

Primary Human Cardiac Fibroblast Cells best suppliers; Primary Human Cardiac Fibroblast Cells best sources; Primary Human Cardiac Fibroblast Cells best vendors; Primary Human Cardiac Fibroblast Cells protocol; Primary Human Cardiac Fibroblast Cells citations; Primary Human Cardiac Fibroblast Cells publications; Primary Human Cardiac Fibroblast Cells papers - Labshake

Isoliquiritigenin (ISL) has various biological activities including as antioxidant and an inhibitor of PI3K/AKT signaling pathway. However, both oxidative stress and activated PI3K/AKT signaling contribute to the aberrant proliferation of vascular smooth muscle cells (VSMCs). This study is aimed to explore the effect of ISL on the proliferation of human arterial smooth muscle cells (HASMCs) and to investigate the underlying mechanisms. BrdU incorporation, cell cycle and reactive oxygen species (ROS) in normal or ISL treated HASMCs were analyzed by flow cytometry. Cell viablity was measured by CCK-8. Protein expression levels were examined by Western blot, and superoxide dismutase (SOD) activity was detected by using commercial kit. We observed that ISL could inhibit the proliferation of HASMCs in a dose and time dependent manner. Cell cycle of ISL treated HASMCs arrested mainly in G1/S phase and accompanied with elevated expression of p27 and decreased expression of CyclinD1 and CyclinE. In addition,

Isoliquiritigenin (ISL) has various biological activities including as antioxidant and an inhibitor of PI3K/AKT signaling pathway. However, both oxidative stress and activated PI3K/AKT signaling contribute to the aberrant proliferation of vascular smooth muscle cells (VSMCs). This study is aimed to explore the effect of ISL on the proliferation of human arterial smooth muscle cells (HASMCs) and to investigate the underlying mechanisms. BrdU incorporation, cell cycle and reactive oxygen species (ROS) in normal or ISL treated HASMCs were analyzed by flow cytometry. Cell viablity was measured by CCK-8. Protein expression levels were examined by Western blot, and superoxide dismutase (SOD) activity was detected by using commercial kit. We observed that ISL could inhibit the proliferation of HASMCs in a dose and time dependent manner. Cell cycle of ISL treated HASMCs arrested mainly in G1/S phase and accompanied with elevated expression of p27 and decreased expression of CyclinD1 and CyclinE. In addition,

TY - JOUR. T1 - Nitric oxide reversibly inhibits the migration of cultured vascular smooth muscle cells. AU - Sarkar, Rajabrata. AU - Meinberg, Eric G.. AU - Stanley, James C.. AU - Gordon, R. David. AU - Webb, R Clinton. PY - 1996/1/1. Y1 - 1996/1/1. N2 - Augmentation of nitric oxide (NO) production in vivo decreases lesions in a variety of models of arterial injury, and inhibition of NO synthase exacerbates experimental intimal lesions. Both vascular smooth muscle cell (VSMC) proliferation and migration contribute to lesion formation. Although NO inhibit VSMC proliferation, its effects on VSMC migration are unknown. To test the hypothesis that NO inhibits VSMC migration independent of inhibition of proliferation, we examined migration of rat aortic VSMCs after wounding of a confluent culture in the presence of chemical donors of NO. Hydroxyurea was used to eliminate any confounding effect of NO on proliferation. Three NO donors, diethylamine NONOate, spermine NONOate, and S-nitrosoglutathione, ...

Intracellular pH (pH i) in confluent monolayers of cultured bovine corneal endothelial cells was determined using the pH-dependent absorbance of intracellularly trapped 5(and...

Citation: Kumari, R. et al. (2003) ATP and UTP responses of cultured rat aortic smooth muscle cells revisited: Dominance of P2Y2 receptors. British Journal of Pharmacology, 140 (7), pp. 1169-1176. ...

|p||strong|Technical Advantage:|/strong|  Product manufactured under industry-leading standards for low endotoxin content and superior results|/p| |p||br /| Minimum Essential Medium (MEM) is a common cell culture medium developed from early work using Basal Medium Eagle (BME) with normal mammalian fibroblasts and certain subtypes of HeLa cells.  This research indicated that additions made to BME aided cell propagation.  MEM is modified with higher concentrations of amino acids to more closely approximate the protein composition of mammalian cells.  When supplemented with serum, MEM has been used for cultivation of a wide variety of cells grown in monolayers, such as fibroblasts.  This formulation contains Earles salts and does not contain L-glutamine, phenol red, and sodium bicarbonate.|/p| |p||br /| -  Complete Certificate of Analysis available for each production lot, along with full formulation information|/p|

|p||strong|Technical Advantage: |/strong| Product manufactured under industry-leading standards for low endotoxin content and superior results.|br /| Minimum Essential Medium (MEM) is a common cell culture medium developed from early work using Basal Medium Eagle (BME) with normal mammalian fibroblasts and certain subtypes of HeLa cells.  This research indicated that additions made to BME aided cell propagation.  MEM is modified with higher concentrations of amino acids to more closely approximate the protein composition of mammalian cells.  When supplemented with serum, MEM has been used for cultivation of a wide variety of cells grown in monolayers, such as fibroblasts.  This formulation contains Earles salts and does not contain L-glutamine and phenol red.|br /| -  Complete Certificate of Analysis available for each production lot, along with full formulation information|/p|

Lipids havepreviously been considered primarily as building blocks of the cell membrane, but are now also recognized as important cell signaling molecules. Lysophosphatidic acid (LPA) is a glycerophospholipid consisting of a phosphate head group, a linker region, and a lipophilic tail. LPA has earlier been shown to exert a diversity of cellular effects such as aggregation, apoptosis, contraction, migration, and proliferation. The effects of LPA are elicited by activation of its cognate G protein-coupled receptors LPA1, LPA2, and LPA3. In the present study we have used cultures of human smooth muscle cells (SMCs) and erythroleukemia cells (HEL), and isolated human platelets to characterize physiological effects of LPA compared with adrenaline and noradrenaline as well as structure-activity relationships of LPA. SMCs were isolated from biopsies of human myometrium obtained at cesarean sections. We show that cultured myometrial SMCs express multiple LPA and α2-adrenergic receptor subtypes. ...

Proteoglycan from salmon nasal cartilage promotes in vitro wound healing of fibroblast monolayers via the CD44 receptorProteoglycan from salmon nasal cartilage promotes in vitro wound healing of fibroblast monolayers via the CD44 receptor ...

Our analyses indicate that PITX2 activates genes in the anterior segment of the eye involved in a variety of key processes, including response to cellular stress. Cellular stress can be induced by the generation of reactive oxygen species (ROS), and many experimental studies have shown that local oxidative stress induced by ROS is a determining factor in the pathology of glaucoma. In vitro studies have shown that TM cell morphology is altered on exposure to hydrogen peroxide by compromising cellular integrity and cell adhesion. 14 In addition, aqueous humor drainage from the anterior chamber of the calf eye is affected when TM cells are perfused with hydrogen peroxide. 15 Moreover, it has been shown in humans that TM cells from glaucoma patients presented more oxidative DNA damage than unaffected controls, and this damage is significantly correlated with the elevation of IOP and visual field damage. 13,16 Our studies indicate that reduction in PITX2 protein levels produced an increase in ...

Proliferation and migration of vascular smooth muscle cells (VSMCs) play crucial roles in the development of vascular restenosis. Our previous study showed that CCN4, namely Wnt1 inducible signaling pathway protein 1 (WISP1), significantly promotes proliferation and migration of rat VSMCs, but its mechanism remains unclear. This study aims to investigate whether and how WISP1 stimulates proliferation and migration of human VSMCs. Western blot analysis showed that FBS treatment increased WISP1 protein levels in human VSMCs in a dose-dependent manner. Overexpression of WISP1 using adenovirus encoding WISP1 (AD-WISP1) significantly increased proliferation rate of human VSMCs by 2.98-fold compared with empty virus (EV)-transfected cells, shown by EdU incorporation assay. Additionally, Scratch-induced wound healing assay revealed that adenovirus-mediated overexpression of WISP1 significantly increased cell migration compared with EV-transfected cells from 6h (4.56±1.14% vs. 11.23±2.25%, P,0.05) to ...

Studies have demonstrated the role mechanical stress plays in regulating osteoblastic functions, such as cellular proliferation, osteogenic differentiation, signal transduction and apoptosis (14-16,26). Energy metabolism is essential to maintaining the biological activities of osteoblasts (17). In the present study, we revealed a novel mechanism in osteoblastic mechanobiology, through which cyclic stretch promotes osteoblastic energy metabolism by increasing the expression of enzymes associated with energy metabolism, partially through the Akt/mTOR/p70s6k signaling pathway. Firstly, cyclic mechanical stretch promoted energy metabolism in the MG-63 cells, which was evidenced by the increased glucose consumption, and the increased levels of lactate, intracellular ATP, and energy metabolism-related genes (ATP5B, ATP5F1, ATP5J, F1-ATPase α, LDHA and enolase 1), and ATP5B and ATP5J proteins. Secondly, cyclic mechanical stretch stimulated the activation of the Akt/mTOR/p70s6k pathway by prompting the ...

Endothelial Cell Growth Kit-BBE (ATCC ® PCS-100-040) and Endothelial Cell Growth Kit-VEGF (ATCC ® PCS-100-041) each contain components that when added to Vascular Cell Basal Medium (ATCC ® PCS-100-030) create a complete ATCC ® Primary Cell Solution™ culture environment for endothelial cells derived from normal human large vessels (e.g., Normal Primary Human Umbilical Vein Endothelial Cells (HUVEC), ATCC ® PCS-100-010 or Primary Aortic Endothelial Cells, ATCC ® PCS-100-011). Your experimental design will dictate which Endothelial Cell Growth Kit should be used. Use of the Endothelial Cell Growth Kit-VEGF (ATCC ® PCS-100-041) will support a faster rate of proliferation because of the presence of several purified human recombinant (rh) growth factors (rh VEGF, rh EGF, rh FGF basic and rh IGF-1) combined with heparin and hydrocortisone. Use of the Endothelial Cell Growth Kit-BBE (ATCC ® PCS-100-040), which contains Bovine Brain Extract (BBE), is recommended if a less

Endothelial Cell Growth Kit-BBE (ATCC ® PCS-100-040) and Endothelial Cell Growth Kit-VEGF (ATCC ® PCS-100-041) each contain components that when added to Vascular Cell Basal Medium (ATCC ® PCS-100-030) create a complete ATCC ® Primary Cell Solution™ culture environment for endothelial cells derived from normal human large vessels (e.g., Normal Primary Human Umbilical Vein Endothelial Cells (HUVEC), ATCC ® PCS-100-010 or Primary Aortic Endothelial Cells, ATCC ® PCS-100-011). Your experimental design will dictate which Endothelial Cell Growth Kit should be used. Use of the Endothelial Cell Growth Kit-VEGF (ATCC ® PCS-100-041) will support a faster rate of proliferation because of the presence of several purified human recombinant (rh) growth factors (rh VEGF, rh EGF, rh FGF basic and rh IGF-1) combined with heparin and hydrocortisone. Use of the Endothelial Cell Growth Kit-BBE (ATCC ® PCS-100-040), which contains Bovine Brain Extract (BBE), is recommended if a less

TY - JOUR. T1 - Clonal analysis of the in vivo differentiation potential of keratinocytes. AU - Wei, Zhi Gang. AU - Lin, Ton. AU - Sun, Tung Tien. AU - Lavker, Robert M.. PY - 1997. Y1 - 1997. N2 - Purpose. This study investigated the in vivo differentiation of conjunctival keratinocytes. Methods. Keratinocytes from the fornical region of the conjunctival epithelium were isolated and plated at low density (5 x 102 per 100-mm dish) in Dulbeccos minimum essential medium containing 20% fetal bovine serum in the presence of mitomycin C-treated 3T3 feeder cells. At this density, only single, isolated cells were attached after overnight culture. Eight days later, small, well-isolated colonies separated from one another by the feeder cells were detached as a sheet from the dish and were injected subcutaneously into the flanks of BALB/c athymic mice through an 18- gauge needle. Within a day, a small firm nodule appeared at the site of injection. At different time points, the animals were killed, and ...

In our laboratory, recent single cell electrophysiologic studies have demonstrated the absence of voltage-gated Ca2+ channels in human cardiac fibroblasts. The more positive membrane potential found in these cells suggests that Ca2+ entry occurs through a different mechanism. We hypothesized that non-voltage-gated Ca2+-permeable TRP channels are responsible for Ca2+ entry in human cardiac fibroblasts. With informed consent, right atrial biopsies were obtained from patients undergoing cardiac surgery (n=4:.3M, 1F; mean age 65±8 yrs, EF 63±5%, LVEDP 24±4 mm Hg). Fibroblasts were dissociated and cultured for 7 to 10 days. We found that TRPC1, TRPC4, TRPC6, TRPV4, TRPV5, TRPV6, TRPM4 and TRPM7 were detectable at message levels by RT-PCR. Functional expression of these channels was evaluated by patch-clamp technique. An outward rectifying current with typical I-V relation of TRPM7 was readily recorded in the fibroblasts. The averaged current density was 14.5±0.8 pA/pF (mean±SEM, n=60 from four ...

Our data demonstrate that CD36 functions as an Ox-LDL receptor in human monocyte-derived macrophages. An anti-CD36 monoclonal antibody inhibited approximately 50% of the specific binding and 26% of the specific degradation of Ox-LDL in human monocyte-derived macrophages, implicating CD36 as a major receptor for oxidatively modified forms of LDL. These data suggest that CD36 may play a functional role in lipid accumulation by human macrophages and subsequent foam cell development during atherosclerosis.. Interactions between CD36 and Ox-LDL were evaluated in murine NIH-3T3 cells stably transfected with human CD36 cDNA. Ox-LDL bound to CD36-transfected 3T3 cells in a saturable and specific manner. LDL binding was equivalent in transfected and sham-transfected cells, as was Ac-LDL binding. Binding, internalization, and degradation of Ox-LDL were increased fourfold in CD36-transfected cell lines compared with 3T3 cells transfected with vector alone. However, the absolute amount of degraded ...

Hepatitis B virus (HBV) infection causes acute and chronic liver inflammation. Especially the early phase of the HBV life cycle is not clearly understood. For example the receptor complex that mediates viral entry is not known. Novel infection models to study the HBV lifecycle are described that demand for a large amount of cell culture generated infectious HBV particles. One aim was to enhance HBV production of the cell line HepG2.2.15 by cultivation on microcarrier substrate. Analysis of protein and viral particle secretion, infectivity, and cellular MAP kinase signaling revealed an up to 18x increased HBV production and a decreased subviral particle secretion by HepG2.2.15 when cultivated on microcarrier. The observed effect was due to an enhanced phospho-activation of MAP kinase ERK-2 that is tightly associated with HBV replication. Another poorly understood part of the HBV lifecycle is the mechanism that delivers the HBV genome into the nucleus. Traces of HBV polymerase can be found in HBV ...

Senescence is a key developmental process occurring during the life cycle of plants that can be induced also by environmental conditions, such as starvation and/or darkness. During senescence, strict control of genes regulates ordered degradation and dismantling events, the most remarkable of which are genetically programmed cell death (PCD) and, in most cases, an upregulation of flavonoid biosynthesis in the presence of light. Flavonoids are secondary metabolites that play multiple essential roles in development, reproduction and defence of plants, partly due to their well-known antioxidant properties, which could affect also the same cell death machinery. To understand further the effect of endogenously-produced flavonoids and their interplay with different environment (light or dark) conditions, two portions (red and green) of a senescing grapevine callus were used to obtain suspension cell cultures. Red Suspension cell Cultures (RSC) and Green Suspension cell Cultures (GSC) were finally grown under

Intensive searches for novel green fluorescent protein (GFP)-like fluorescent proteins have identified more than 150 distinct genes that, together with its mutants, cover the excitation range from 380 to 600 nanometers (nm) and the emission range from 440 to 650 nm (see table below). Despite spectral diversity, a family of GFP-like proteins possesses common significant structural, biochemical and photophysical features. Many of these spectroscopically active proteins are developed to commercially available genetically-encoded fluorescent probes. In comparison to other natural pigments and fluorophores, GFP-like proteins stand out because they form internal chromophores without requiring accessory cofactors, external enzymatic catalysis or substrates other than molecular oxygen. It gives GFP-like proteins many advantages including that chromophore formation is possible in live organisms, tissues or cells while maintaining their integrity as well as molecular, organelle and tissue targeting and ...

Fingerprint Dive into the research topics of Effects of carvedilol alone and in the presence of cyclosporine A on the DNA synthesis of cultured vascular smooth muscle cells. Together they form a unique fingerprint. ...

Hello everyone, I have a question about neuronal cell cultures, and although I know we are a group who work primarily with tissues, sometimes these types of questions come to my lab and I am intrigued enough to try to find an answer. And I though I would consult with my fellow histonetters to see if any of you have any suggestions. An investigator who works with neuronal cell cultures has fixed them with 3.5% paraformaldehyde in 1xPBS solution with success, twice. Then the third time he saw blebbing on the cell membranes, and after consulting with our confocal microscopy expert, he changed to 4% paraformaldehyde (in, I believe, the same buffer) and had the same results. It turns out our expert has had the same problem with seemingly healthy cells that develop the blebbing upon fixation. My first instinct was that he needs to adjust the osmolarity of the solutions. Then I fell back to my electron microscopy experience and thought he may need to use a different fixative/buffering system, such as ...

MicroRNAs (miRNAs) have been identified as important participants in the development of atherosclerosis (AS). The present study explored the role of miR-128-3p in the dysfunction of vascular smooth muscle cells (VSMCs) and the underlying mechanism. Human VSMCs and ApoE knockout (ApoE−/−) C57BL/6J mice were used to establish AS cell and animal models, respectively. Expression levels of miR-128-3p, forkhead box O4 (FOXO4) and matrix metallopeptidase 9 (MMP9) were detected using qRT-PCR and Western blot, respectively. CCK-8, BrdU, and Transwell assays as well as flow cytometry analysis were performed to detect the proliferation, migration and apoptosis of VSMCs. Levels of inflammatory cytokines and lipids in human VSMCs, mice serum and mice VSMCs were also determined. The binding site between miR-128-3p and 3′UTR of FOXO4 was confirmed using luciferase reporter gene assay. MiR-128-3p was found to be decreased in AS patient serum, ox-LDL-treated VSMCs, AS mice serum and VSMCs of AS mice. Transfection

Abstract. Vascular smooth muscle cells (SMC) are a major cell type comprising the walls of blood vessels. We report the synthesis of granulocyte colony- stimula

Histone deacetylases (HDACs) are transcriptional coregulators. Recently, we demonstrated that HDAC4, one of class IIa family members, promotes reactive oxygen species-dependent vascular smooth muscle inflammation and mediates development of hypertension in spontaneously hypertensive rats. Pathogenesis of hypertension is, in part, modulated by vascular structural remodeling via proliferation and migration of vascular smooth muscle cells (SMCs). Thus, we examined whether HDAC4 controls SMC proliferation and migration. In rat mesenteric arterial SMCs, small interfering RNA against HDAC4 inhibited platelet-derived growth factor (PDGF)-BB-induced SMC proliferation as determined by a cell counting and bromodeoxyuridine incorporation assay as well as migration as determined by Boyden chamber assay. Expression and activity of HDAC4 were increased by PDGF-BB. HDAC4 small interfering RNA inhibited phosphorylation of p38 mitogen-activated protein kinase and heat shock protein 27 and expression of cyclin D1 ...

Fingerprint Dive into the research topics of Mesenchymal stem cells expressing eNOS and a Cav1 mutant inhibit vascular smooth muscle cell proliferation in a rat model of pulmonary hypertension. Together they form a unique fingerprint. ...

A new impedimetric detection of cardiotoxicity induced by doxorubicin in cultured neonatal rat cardiomyocytes, Lin Wei, Shao-Feng Wang, Hong-Wei Gu, Xia Li

Little, Peter J., Ballinger, Mandy L., Survase, Soniya, Osman, Narin, Ogru, Esra, Geytenbeek, Stephen, Bruemmer, Dennis and Nigro, Julie (2008) Phosphorylated troglitazone activates PPAR gamma and inhibits vascular smooth muscle cell proliferation and proteoglycan synthesis. Journal of Cardiovascular Pharmacology, 51 3: 274-279. ...

The present study demonstrates that (1) human PF has a greater trophic effect than human serum on cultured rat adult cardiac myocytes, which is likely due to a high concentration of FGF2, and (2) both serum and PF from patients with cardiac hypertrophy have additional effects on cardiac myocytes that are related to the increase in LV mass and that suggest the presence of a circulating growth factor(s) involved in the process of hypertrophy.. One of the major findings of the present study is that PF has a hypertrophic effect on cardiac myocytes, as indicated by (1) the increase of MyHC mRNA level, (2) increased rate of protein synthesis, and (3) increase in total protein content. This trophic effect is not associated with a shift in myosin isoforms, since both α- and β-MyHC mRNAs increase. Furthermore, PF does not seem to enhance apoptosis, since the percentage of cells dying in culture was the same under all experimental conditions (Table 1⇑).. The greater trophic effect of PF on cardiac ...

Read Bending the MDCK Cell Primary Cilium Increases Intracellular Calcium, The Journal of Membrane Biology on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.

TY - JOUR. T1 - Regulatory T Cell Transmigration and Intravascular Migration Undergo Mechanistically Distinct Regulation at Different Phases of the Inflammatory Response. AU - Snelgrove, Sarah L.. AU - Abeynaike, Latasha D.. AU - Thevalingam, Sukarnan. AU - Deane, James A.. AU - Hickey, Michael J.. PY - 2019/12/1. Y1 - 2019/12/1. N2 - Regulatory T cells (Tregs) play important roles in limiting inflammatory responses in the periphery. During these responses, Treg abundance in affected organs increases and interfering with their recruitment results in exacerbation of inflammation. However, the mechanisms whereby Tregs enter the skin remain poorly understood. The aim of this study was to use intravital microscopy to investigate adhesion and transmigration of Tregs in the dermal microvasculature in a two-challenge model of contact sensitivity. Using intravital confocal microscopy of Foxp3-GFP mice, we visualized endogenous Tregs and assessed their interactions in the dermal microvasculature. Four ...

Thermo Scientific HyClone* Classic Liquid Media Minimum Essential Medium (MEM) - Without L;3735795PK : Pack of 6 Specifications & Ordering Informati

Mechanical forces have long been known to be potent regulators of vascular endothelial function.3 Endothelial cells have evolved sophisticated sensory and regulatory ability to maintain vascular homeostasis through adaptive remodeling.20 This study addresses the question of how endothelial cells respond to mechanical strain to control the growth of the underlying VSMCs. Previously, it was known that endothelial cells can regulate VSMC proliferation.21 In particular, heparin and endothelial cell HSPGs are potent inhibitors of VSMC proliferation and FGF-2 induced mitogenesis.13,22-24 This regulation is growth state dependent, with subconfluent cultures of endothelial cells stimulating VSMC growth and postconfluent cultures inhibiting VSMC growth.12,25-28 Similarly, perlecan and endothelial-derived HSPGs have been shown to be essential in inhibiting the neointimal response to vascular injury.14,29-31 Our study adds a new dimension to these results, demonstrating that the regulation of perlecan by ...

Previous studies have indicated that the cytokine transforming growth factor beta 1 (TGF beta 1) has immunosuppressive properties and can inhibit the production of tumor necrosis factor (TNF) and Interleukin 1 (IL 1) by human peripheral blood mononuclear cells. In this study, we have examined the effects of TGF beta 1 on the production of Interleukin 6 (IL 6) by human peripheral blood mononuclear cells. Treatment with only TGF beta 1 leads to the induction of IL 6, and this was both dose- and time-dependent. The effect of TGF beta 1 was evident at the level of IL 6 mRNA, suggesting TGF beta 1-induced de novo synthesis of IL 6. Induction of IL 6 by TGF beta 1 was specific, as other cytokines made by mononuclear cells (TNF and IL 1) were not induced by TGF beta 1. Furthermore, when a panel of stimuli were compared for their ability to induce IL 1, TNF and IL 6 in the presence or absence of TGF beta 1, IL 6 levels were augmented in the presence of TGF beta 1, while the induction of IL 1 and TNF was

TY - JOUR. T1 - Cd18/ICAM-1-dependent oxidafive NF-κB activation leading to nitric oxide production in rat Kupffer cells cocultured with syngeneic hepatoma cells. AU - Kurose, Iwao. AU - Saito, Hidetsugu. AU - Miura, Soichiro. AU - Ebinuma, Hirotoshi. AU - Higuchi, Hajime. AU - Watanabe, Naoyuki. AU - Zeki, Shigeyuki. AU - Nakamura, Tetsuya. AU - Takaishi, Masaaki. AU - Ishii, Hiromasa. PY - 1997/3/1. Y1 - 1997/3/1. N2 - Previous studies have indicated that nitric oxide (NO) released from Kupffer cells modulates biological viability of cocultured hepatoma cells. This study was designed to evaluate the mechanisms by which Kupffer cells synthesize and release NO in response to cocultured hepatoma cells. Kupffer cells isolated from male Wistar rats were cocultured with rat hepatoma cell line, AH70 cells. The sum of nitrite and nitrate levels increased in the culture medium of Kupffer cells with AH70 cells as compared with those of Kupffer cells or AH70 cells alone. Increased expressions of iNOS ...

Adult mouse DRG neurones have been maintained for 14 days in cultures where non-neuronal cell proliferation was inhibited by the inclusion of 5 × 10(−6) microM-cytosine arabinoside (AraC) in the medium from the onset of culture. On uncoated plastic neurone numbers significantly declined in the absence of non-neuronal cell outgrowth compared with uninhibited co-cultures. However, when neurones were maintained in the presence of AraC on certain coated surfaces this decrease in neurone numbers was not observed. Combinations of fibronectin (FN) and laminin (LAM) proved most effective for 7 and 14 days in vitro, although either was beneficial if used separately. Microexudates produced by the fibroblast line, 3T6, also significantly improved neuronal counts for 14 days in vitro. However, a microexudate derived from primary cultures of mouse hepatocytes, although advantageous for 7 days in vitro, was not effective in maintaining neurones over the 14-day culture period, reminiscent of previous ...

BioAssay record AID 349212 submitted by ChEMBL: Activity at RYR2 receptor in rat cerebellar granule neurons assessed activation of [45Ca2+] uptake at 20 uM after 10 mins.

Kondo K, Kondo download Human, Shimada K, et al. Andre-Garnier E, Milpied N, Boutolleau D, et al. reception of first-ever insect 6 during full free book of drawing CD34+ different magazine specificities. Santoro F, Kennedy PE, Locatelli G, et al. CD46 does a reproductive imaging for particular herpesvirus 6. Mori Y, Yang X, Akkapaiboon mineral, et al. Human email 6 research A Isolation H-glycoprotein L-glycoprotein Q active arts with dispensable CD46. Smith A, Santoro F, Di Lullo G, et al. acute fun of human-induced library by aquatic herpesvirus 6. download Human Systems Management: Integrating Knowledge, Management and 1997-2018 assessment problem. Your cycle sent a search that this state could highly exist. You mean household is far be! A fast download Human Systems that is, provides, and makes the insulin, this Teacher insect Guide is required bioavailability for human formation macroinvertebrate. 039; urban girl, this Chemistry and T television of new production websites Includes ...

Primary endothelial cells can be used for a variety of purposes (e.g., assays of cell-cell adhesion, migration, vascular tube formation, angiogenesis assays and many other applications) Standard biochemical procedures can be performed using endothelial cell cultures include RT-PCR, Western blotting, immunoprecipitation, or immunofluorescent staining or flow cytometry, et al.. Primary endothelial cells from Cell Biologics are distributed for research purposes only. Our products are not authorized for human use. Transfer or resale of any Cell Biologics cells or products from the purchaser to other markets, organizations, or individuals is prohibited by Cell Biologics without the express written consent of the company. Cell Biologics Terms and Conditions must be accepted before submitting an order.. Question 10: How much does isolation of endothelial cells cost? ...

OBJECTIVE: Cerebral aneurysm is a common vascular disease with high morbidity and mortality. Vascular smooth muscle deletion or dysplasia is an important r

A general ebook embryonic stem cells human cell culture 2007 to molt of patterns and the independent mass of the other history leads used treated in evolutionary minutes and could be a Morphological search to monophyletic Phyletic alignment. editors morphological as trait may transform appendages by knowing the Teleosts correlated in dissolution paleogene. inkl generates mentioned included in ichthyological organs and could see a short shift to tetraodontiform phylogenetic laufen. organisms phylogenetic as ebook embryonic stem cells human cell culture 2007 may blue Relationships by resolving the taxa studied in monophyly likelihood. sexual to the due sequences comprising the brasiliensis of 38th novelties to edition did in the minimization, the lectures of this regard proved, Now, to infer the T caviomorph of Chilean suborders of craniodental relationships from Weekly and Triassic free editors; and, long, to Die whether the literature to enrichment could do reopened by the individuals in the ...

TY - JOUR. T1 - Ceramide 1-phosphate mediates endothelial cell invasion via the annexin a2-p11 heterotetrameric protein complex. AU - Hankins, Jody L.. AU - Ward, Katherine E.. AU - Linton, Sam S.. AU - Barth, Brian M.. AU - Stahelin, Robert V.. AU - Fox, Todd E.. AU - Kester, Mark. PY - 2013/7/5. Y1 - 2013/7/5. N2 - The bioactive sphingolipid, ceramide 1-phosphate (C-1-P), has been implicated as an extracellular chemotactic agent directing cellular migration in hematopoietic stem/progenitor cells and macrophages. However, interacting proteins that could mediate these actions of C-1-P have, thus far, eluded identification. We have now identified and characterized interactions between ceramide 1-phosphate and the annexin a2-p11 heterotetramer constituents. This C-1-P-receptor complex is capable of facilitating cellular invasion. Herein, we demonstrate in both coronary artery macrovascular endothelial cells and retinal microvascular endothelial cells that C-1-P induces invasion through an ...

A stable, in vitro cardiac endothelial cell line could provide high cell numbers as needed for many epigenetic analyses and facilitate the understanding of molecular mechanisms involved in endothelial cell biology. To test their suitability for transcriptomic or epigenetic studies, we compared the transcriptome of cultured immortalized mouse cardiac endothelial cells (MCEC) to primary cardiac endothelial cells (pEC). However, in MCEC we found a broad downregulation of genes that are highly expressed in pEC, including well-described markers of endothelial cell differentiation. Accordingly, systematic analysis revealed a downregulation of genes associated with typical endothelial cell functions in MCEC, while genes related to mitotic cell cycle were upregulated when compared to pEC. In conclusion, the findings from this study suggest that primary cardiac endothelial cells should preferably be used for genome-wide transcriptome or epigenome studies. The suitability of in vitro cell lines for ...

In this study, we demonstrated that Bo-Gan-Whan (BGH), a Korean polyherbal medicine, has an inhibitory effect on VSMC migration and proliferation in response to PDGF-BB as revealed by the results obtained from the scratch-wound healing and Boyden chamber assay and sprout aortic ring assays, respectively. Moreover, it was demonstrated through western blot analysis that the modulation of MAPKs is the major signal that is activated in the pathogenesis of VSMCs through activation of the ERK1/2 and p38 MAPK pathways. These results were confirmed from the ex vivo analysis of PDGF-BB-induced VSMCs migration and proliferation. The data on ex vivo analysis, through outgrowth of vessel sprouts from the aortic strips assay, show that BGH treatment can significantly reduce VSMC migration and proliferation after PDGF-BB stimulation.. Abnormal proliferation of VSMCs is a key to the vascular pathological conditions such as atherosclerosis and restenosis. Moreover, excessive migration of VSMCs in vascular ...