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 ...
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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
Cellular implantation is an emerging therapy for repair of the injured myocardium; however, the mechanisms of improvement and the optimal donor cell type remain unknown. We designed a reproducible, well-controlled in vitro assay for comparing the efficacy of different donor cell types to electrically couple with cardiomyocytes and propagate action potentials. Using soft lithography, we micropatterned 100um-wide strands of neonatal rat ventricular myocytes with an empty insert region of controlled length for donor cell attachment (implantation). Insert lengths were confirmed by immunostaining. Electrical conduction of Ca2+ transients or membrane voltage was optically mapped at 504 sites spaced 37um. The conduction time (CT) between two 638um spaced recording sites was measured in pure cardiac strands (control) and across inserts populated with different donor cell types. Control cardiac strands produced a CT of 2ms (conduction velocity of 30cm/s). Skeletal myoblasts and wild type HEK-293 cells, ...
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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 ...
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TY - JOUR. T1 - The effect of anti-hypertensive drugs on DNA synthesis and proliferation of cultured rat aortic smooth muscle cells. AU - Kang, Shin Wook. AU - Lee, In Hee. AU - Choi, Kyu Hun. AU - Lee, Ho Yung. AU - Han, Dae Suk. PY - 1997. Y1 - 1997. N2 - The aim of this study was to elucidate the effects of anti-hypertensive drugs, nifedipine, furosemide, hydrochlorothiazide, captopril, and atenolol on DNA synthesis and proliferation of cultured rat aortic smooth muscle cells induced by fetal calf serum. Aortic smooth muscle cells from Sprague-Dawley rats were isolated, cultured, and seeded in multi-well plates. When confluent, cells were cultured in a conditioned medium without fetal calf serum. After 72 hours, cells were cultured in the medium retaining 10% fetal calf serum with or without anti-hypertensive drugs by increasing the concentration between 10-8 and 10-4M. DNA synthesis was assessed by [3H]-thymidine uptake and proliferation by cell numbers using a hemocytometer. Nifedipine at a ...
TY - JOUR. T1 - Down-regulation of superoxide dismutase gene expression in cultured rat aortic smooth muscle cells (A7r5) after long-term incubation with vitamin C. AU - Liu, J. C.. AU - Chow, J. M.. AU - Tsai, M. F.. AU - Hsieh, M. H.. AU - Chen, Y. J.. AU - Chan, P.. PY - 2000. Y1 - 2000. N2 - Background: Oxygen free radicals have been linked to the process of cardiovascular disease and aging. Epidemiological studies supported the beneficial effect of supplementation of antioxidants such as vitamin C and vitamin E. Superoxide dismutase (SOD) is a endogenous enzyme system which can scavenge oxygen free radicals. This study investigated the effect of supplementation of ascorbic acid (vitamin C) on the changes of SOD. Methods: Rat aortic smooth muscle cells (A7r5) were divided into 4 groups: a control group (without vitamin C) and treatment groups with vitamin C at 50 μM, 100 μM and 200 μM. After a short-term (2 days) or long-term (7 days) incubation, the enzyme activity and mRNA level of SOD ...
Hello. I need to culture bovine aortic smooth muscle cell. I do not know what medium should be used. What supplements or growth factors are required? In how much in amount? Please give me some help. Thanks in advance ...
TY - JOUR. T1 - TLR3-mediated synthesis and release of Eotaxin-1/CCL11 from human bronchial smooth muscle cells stimulated with double-stranded RNA. AU - Niimi, Kyoko. AU - Asano, Koichiro. AU - Shiraishi, Yoshiki. AU - Nakajima, Takeshi. AU - Wakaki, Misa. AU - Kagyo, Junko. AU - Takihara, Takahisa. AU - Suzuki, Yusuke. AU - Fukunaga, Koichi. AU - Shiomi, Tetsuya. AU - Oguma, Tsuyoshi. AU - Sayama, Koichi. AU - Yamaguchi, Kazuhiro. AU - Natori, Yukikazu. AU - Matsumoto, Misako. AU - Seya, Tsukasa. AU - Yamaya, Mutsuo. AU - Ishizaka, Akitoshi. PY - 2007/1/1. Y1 - 2007/1/1. N2 - Respiratory infections with RNA viruses, such as rhinovirus or respiratory syncytial virus, are a major cause of asthma exacerbation, accompanied by enhanced neutrophilic and/or eosinophilic inflammation of the airways. We studied the effects of dsRNA synthesized during RNA virus replication, and of its receptor, TLR3, on the synthesis of eosinophilic chemokines in bronchial smooth muscle cells (BSMC). Synthetic dsRNA, ...
TY - JOUR. T1 - Long-term zinc deprivation accelerates rat vascular smooth muscle cell proliferation involving the down-regulation of JNK1/2 expression in MAPK signaling. AU - Alcantara, Ethel H.. AU - Shin, Mee Young. AU - Feldmann, Jörg AU - Nixon, Graeme F.. AU - Beattie, John H.. AU - Kwun, In Sook. PY - 2013/5/1. Y1 - 2013/5/1. N2 - Background: The accelerated proliferation of vascular smooth muscle cells (VSMCs) is a contributor for atherosclerosis by thickening the vascular wall. Since zinc modulation of VSMC proliferation has not been clarified, this study investigated whether zinc affects VSMC proliferation. Methods and results: Both a rat aorta origin vascular smooth muscle cell line (A7r5 VSMCs) and primary VSMCs which were collected from rat aorta (pVSMCs) were cultured with zinc (0-50 µM Zn) for short- (=12 d) and long-term (28 d) periods under normal non-calcifying (0 or 1 mM P) or calcifying (,2 mM P) P conditions. Mouse vascular endothelial cells (MS I cells) were also cultured ...
PURPOSE. A previous report has described an ocular surface reconstruction method involving the use of cultivated corneal epithelium derived from limbal explants. In the current study, a new culture system was developed involving the in vitro propagation on amniotic membrane (AM) of epithelial cells from enzymatically dissociated limbal epithelium. The purpose of this new method is to produce a cultivated epithelial cell layer that contains stem cells and that is superior to explanted cultivated epithelium. The new cell-suspension technique was compared with the existing explant method. METHODS. Limbal epithelial cells were dissociated from donor eyes by dispase and seeded on the denuded AM. Small pieces of limbal epithelium were also cultured on denuded AM as explant cultures. The cultivated epithelium was examined by electron microscopy and immunohistochemistry for cornea-specific keratins (K3 and K12). RESULTS. Both cell-suspension and explant culture methods produced a healthy epithelial cell ...
PubMed journal article: Comparison of intracellular calcium signals evoked by heat and capsaicin in cultured rat dorsal root ganglion neurons and in a cell line expressing the rat vanilloid receptor, VR1. Download Prime PubMed App to iPhone, iPad, or Android
primary hepatocyte culture - posted in Tissue and Cell Culture: Ill start the human primary hepatocyte culture. I have no idea on the important issue how long primary hepatocyte can maintain its bioactivity. Does anybody has experience on culturing of human primary hepatocytes?
TY - JOUR. T1 - Protein phosphorylation in primary astrocyte cultures treated with and without dibutyryl cyclic AMP. AU - Neary, Joseph T.. AU - Gutierrez, Maria del Pilar. AU - Norenberg, Luz Oliva B.. AU - Norenberg, Michael D.. PY - 1987/4/28. Y1 - 1987/4/28. N2 - Protein phosphorylation was investigated in primary rat astrocyte cultures treated with and without dibutyryl cyclic AMP. Astrocytes maintained in dibutyryl cyclic AMP for several weeks displayed increased phosphate incorporation in 5 protein bands (55, 52, 45, 43 and 28 kDa) while incorporation in one band (42 kDa) was decreased. Phosphate incorporation in several other protein bands was unchanged. Calcium-dependent phosphate incorporation was also altered by prior exposure of the cells to dibutyryl cyclic AMP: addition of calcium to broken cell preparations resulted in increased incorporation in 75, 53 and 52 kDa while decreased incorporation occurred in 100 kDa. These differences in protein phosphorylation may be related to the ...
TY - JOUR. T1 - WISP1, a pro-mitogenic, pro-survival factor, mediates tumor necrosis factor-α (TNF-α)-stimulated cardiac fibroblast proliferation but inhibits TNF-α-induced cardiomyocyte death. AU - Venkatachalam, Kaliyamurthi. AU - Venkatesan, Balachander. AU - Valente, Anthony J.. AU - Melby, Peter. AU - Nandish, Sailesh. AU - Reusch, Jane E B. AU - Clark, Robert A.. AU - Chandrasekar, Bysani. PY - 2009/5/22. Y1 - 2009/5/22. N2 - WNT1-inducible signaling pathway protein-1 (WISP1), a member of the CYR61/CTGF/Nov family of growth factors, can mediate cell growth, transformation, and survival. Previously we demonstrated that WISP1 is up-regulated in post-infarct heart, stimulates cardiac fibroblast proliferation, and is induced by the proinflammatory cytokine tumor necrosis factor-α (TNF-α). Here we investigated (i) the localization of TNF-α and WISP1 in post-infarct heart, (ii) the mechanism of TNF-α-mediated WISP1 induction in primary human cardiac fibroblasts (CF), (iii) the role of ...
Objective(s): To investigate the role of autophagy in advanced glycation end products (AGEs)-induced proliferation and migration in rat vascular smooth muscle cells (VSMCs).Materials and Methods: After culture, VSMCs were treated with 0, 1, 10, and 100 μg/ml concentrations of AGEs. Autophagy specific protein light chain 3 (LC3)-I/II was determined by western blotting, autophagosomes were observed with electron microscopy, cell proliferation was quantified using the methyl thiazolyl tetrazolium (MTT) assay, and cell migration was evaluated using Transwell migration and scratch assays. Results: Compared to the control group, the level of LC3- II/I in AGEs treatment group was up-regulated, and the number of autophagosomes was also increased. Furthermore, in concentration of 100 μg/ml AGEs, the extent of proliferation and migration was significantly increased compared to the control group. However, pretreating cells with autophagy inhibitor 3-MA could attenuate these effects.Conclusion: Our study
Cysteine-containing leukotrienes (cysteinyl-LTs) are pivotal inflammatory mediators that play important roles in the pathophysiology of asthma, allergic rhinitis, and other inflammatory conditions. In particular, cysteinyl-LTs exert a variety of effects with relevance to the aetiology of asthma such as smooth muscle contraction, eosinophil recruitment, increased microvascular permeability, enhanced mucus secretion and decreased mucus transport and, finally, airway smooth muscle cells (ASMC) proliferation. We used human ASMC (HASMC) to identify the signal transduction pathway(s) of the leukotriene D4 (LTD4)-induced DNA synthesis. Proliferation of primary HASMC was measured by [3H]thymidine incorporation. Phosphorylation of EGF receptor (EGF-R) and ERK1/2 was assessed with a polyclonal anti-EGF-R or anti-phosphoERKl/2 monoclonal antibody. A Ras pull-down assay kit was used to evaluate Ras activation. The production of reactive oxygen species (ROS) was estimated by measuring dichlorodihydrofluorescein (DCF
BioAssay record AID 44634 submitted by ChEMBL: HSF produced by bone marrow-derived stromal cell lines C6.4 on stimulation with the compound at (1000 ng/mL) was determined in vitro in an GM-CFC assay..
Purpose: : To investigate the in vitro effects of tobacco-specific n- nitrosamines (TSNA); (N-Nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) which are the major components of cigarette smoke on human retinal pigment epithelial cells. Methods: : Human ARPE-19 cultures were exposed to varying concentrations of NNN and NNK (1mM, 100µM, 10µM, 1 µM) in DMEM/F12 medium for 24 and 48 hours. MTT cell proliferation assays were performed to find a non toxic dose that would not kill the cells within the 24-48 hour period. For both NNN and NNK a 10µM dose of 24 hours was chosen to carry out the rest of the experiments. Total cell protein extracts were analyzed for the presence of OGG1, a DNA repair enzyme and VEGF. Live cells were stained with Hoechst and Mitotracker Green to visualize the nucleus and mitochondrial mass respectively using a confocal microscope. Cells were also labeled for anti- VEGFR2 and imaged using a confocal microscope. Comet assay was performed on ...
TY - JOUR. T1 - Basic fibroblast growth factor-induced prostaglandin production and its intracellular mechanisms in cultured bovine luteal cells. AU - Uenoyama, Yoshihisa. AU - Murakami, Shuko. AU - Schams, Dieter. AU - Okuda, Kiyoshi. N1 - Copyright: Copyright 2017 Elsevier B.V., All rights reserved.. PY - 2000. Y1 - 2000. N2 - The effect of basic fibroblast growth factor (bFGF) on the production of prostaglandin (PG) F2α and PGE2, and the intracellular mechanisms of its action, were investigated in cultured bovine mid-luteal cells (days 8-12 of the estrous cycle). The cells were cultured for 24 h and then exposed to varying concentrations of bovine recombinant bFGF (rbFGF, 1-100 ng/ml) for a further 24 h. A 24-h stimulation with the highest concentration of rbFGF resulted in increases in both PGF2α and PGE2 production by mid-luteal cells (P,0.05). Both U-73122 (an inhibitor of phospholipase (PL) C, 10-6 M) and anthranilic acid (ACA; an inhibitor of PLA2, 10-6 M) inhibited the rbFGF-induced ...
Human pulmonary artery endothelial cells cryopreserved at passage 3. The endothelial cells express von Willebrand factor and are negative for alpha smooth muscle actin. Axol pulmonary artery endothelial cells can be passaged more than 4 times in Artery Endothelial Cell Culture Medium (ax3810). ...
Human Cardiomyocytes Stem Cell Culture Extra-cellular Differentiation Matrix is essential for Differentiation of Human Cardiomyocytes Stem Cell Cultures. This product requires Human Cardiomyocytes Stem Cell Culture Media Cat#M36003-04 and Cells Cat# 36003-04. Also available Products ...
TY - JOUR. T1 - Integrin signaling via the PI3-kinase-Akt pathway increases neuronal resistance to glutamate-induced apoptosis. AU - Gary, Devin S.. AU - Mattson, Mark P.. PY - 2001. Y1 - 2001. N2 - Integrins are integral membrane proteins that mediate adhesive interactions of cells with the extracellular matrix and with other cells. Integrin engagement results in activation of intracellular signaling cascades that effect several different cellular responses including motility, proliferation and survival. Although integrins are known to provide cell survival signaling in various types of non-neuronal cells, the possibility that integrins modulate neuron survival has not been explored. We now report data demonstrating a neuroprotective function of integrins in embryonic hippocampal neurons. Neurons grown on laminin, an integrin ligand, exhibit increased resistance to glutamate-induced apoptosis compared with neurons grown on polylysine. Neurons expressed integrin β1 and treatment of cultures ...
The recognition that cells of the vascular wall can secrete cytokines such as IL-1 suggests new mechanisms for initiating or sustaining inflammatory responses in blood vessels. We report that purified human monocyte-derived IL-1 or recombinant human IL-1 (rIL-1 beta and rIL-1 alpha) induce cultured human smooth muscle cells derived from veins or arteries to synthesize IL-1 beta mRNA and produce and release biologically active IL-1. rIL-1 beta also stimulated the production of PGE2 by smooth muscle cells. Exposure to rIL-1 beta (1-100 ng/ml), or rIL-1 alpha (0.01-10 ng/ml) increased IL-1 beta mRNA levels within 30 min. Actinomycin D (1 microgram/ml) prevented the induction of IL-1 beta mRNA by rIL-1. IL-1 alpha mRNA was detected in SMC treated with cycloheximide (1 microgram/ml) and rIL-1 beta, or cycloheximide alone. rIL-1 alpha and rIL-1 beta produced maximal levels of IL-1 beta mRNA after 4 h, and intracellular IL-1 biological activity after 6 h of exposure. Release of IL-1 activity in the ...
TY - JOUR. T1 - Differential effects of lipoprotein lipase on tumor necrosis factor-α and interferon-γ-mediated gene expression in human endothelial cells. AU - Kota, Rama S.. AU - Ramana, Chilakamarti V.. AU - Tenorio, Fatima A.. AU - Enelow, Richard I.. AU - Rutledge, John C. PY - 2005/9/2. Y1 - 2005/9/2. N2 - Lipoprotein lipase (LPL) is a key enzyme in the hydrolysis of triglyceride-rich lipoproteine. In vascular diseases, such as atherosclerosis, inflammation plays an important role in the pathogenesis of the disease. We examined the role of LPL in modulating tumor necrosis factor-α (TNF-α)- and interferon-γ (IFN-γ)-mediated inflammatory cytokine signal transduction pathways in human aortic endothelial cells (HAECs). LPL significantly suppressed TNF-α-induced gene expression, and this suppression was reversed by tetrahydrolipstatin and heparinase. In contrast, LPL synergistically enhanced IFN-γ-induced gene expression in HAECs. To elucidate the molecular mechanisms of LPL action, we ...
TY - JOUR. T1 - Raman spectroscopy of primary bovine aortic endothelial cells: a comparison of single cell and cell cluster analysis. AU - McManus, L. AU - Boyd, A. AU - Burke, GA. AU - Meenan, BJ. PY - 2011. Y1 - 2011. N2 - There are many techniques that allow in vitro interactions among cells and their environment to be monitored, including molecular, biochemical and immunochemicaltechniques. Traditional techniques for the analysis of cells often require fixation or lysis from substrates; however, use of such destructive methods is not feasible where the expanded cell cultures are required to be used for clinical implantation. Several studies have previously highlighted the potential of Raman spectroscopy to provide useful information on key biochemical markers within cells. As such, we highlight thecapability of Raman spectroscopy with different laser spot sizes for use as a non-invasive, rapid, and specific method to perform in situ analysis of primary bovine aortic endothelial cells ...
TY - JOUR. T1 - Cyclic strain effects on human monocyte interactions with endothelial cells and extracellular matrix proteins. AU - Yun, Jong K.. AU - Anderson, James M.. AU - Ziats, Nicholas P.. PY - 1999. Y1 - 1999. N2 - Human vascular endothelial cells (ECs) are exposed to various levels of hemodynamic forces, cyclic strain, and shear stress in vivo. Here, we examined the in vitro effects of the various levels (0-6%, 7-16%, and 17- 25%) of strain at 60, 30, and 15 cycles per minute (cpm) on human monocyte adherence to endothelial cells and extracellular matrix protein preadsorbed surfaces. Monocyte adhesion to endothelial cells under cyclic strain significantly increased. At both 30 and 60 cpm, ECs under strains of 7-16% and 17-25% showed ,52% and ,117% higher monocyte adhesion than endothelial cells under static condition when monocytes were added for 0.5 h. This increase in monocyte adhesion to ECs under cyclic strain remained significantly higher even after 24 h of incubation. Human ...
P149 Angiotensin (Ang) II acts as a mitogen in vascular smooth muscle cells (VSMC) via multiple signaling mechanisms, including activation of tyrosine kinases and mitogen-activated protein kinase (MAPK). In addition, we have reported that cytosolic phospholipase A2 (cPLA2)-dependent release of arachidonic acid (AA) is linked to Ang II-induced VSMC growth independent of tyrosine kinase. Ang II-mediated activation of cPLA2 involves translocation to the nucleus, as well as serine phosphorylation. Furthermore, VSMC growth stimulated by Ang II has been shown to involve phosphoinositol 3-kinase (PI3K). Thus, Ang II-mediated cPLA2 activation may require a MAPK-dependent phosphorylation, while additional kinases may couple Ang II to the activation of MAPK. Therefore, we used cultured rat VSMC to examine the role of PI3K and MAPK in the phosphorylation of cPLA2, release of AA and growth induced by Ang II. Exposure of VSMC to Ang II (100 nM) increased 3H-thymidine incorporation and cell number by 82% and ...
This study investigated a reciprocal effects model (REM) of childrens body fat self-concept and physical self-concept, and objectively measured school physical activity at different intensities. Grade four students (N = 376; M age = 9.07, SD = .61; 55% boys) from the midwest region of the United States completed measures of physical self-concept and body fat self-concept, and wore accelerometers for three consecutive school days at the beginning and end of one school year. Findings from structural equation modeling analyses did not support reciprocal effects. However, childrens body fat self-concept predicted future physical self-concept and moderate-to-vigorous physical activity (MVPA). Multigroup analyses explored the moderating role of weight status, sex, ethnicity, and sex*ethnicity within the REM. Findings supported invariance, suggesting that the observed relations were generalizable for these children across demographic groups. Links between body fat self-concept and future physical ...
Bone marrow stromal cells protect hematopoietic cells and provide drug resistance by delivering bunch of variable proteins. Thus, alterations of protein expression are typically associated with cell-cell signal transduction and regulation of cellular functions. Co-culture models of bone marrow stromal cells and hematopoietic cells are often used in studies of their crosstalk. Studies of altered protein expression initiated by stromal cell/hematopoietic cell interactions are an important new trend in microenvironmental research. There has been no report to date of global quantitative proteomics analysis of crosstalk between hematopoietic cells and stromal cells. In this study, we analyzed quantitative proteomes in a co-culture system of stromal HS5 cells and hematopoietic KG1a cells, and simultaneously tracked differentially expressed proteins in two types of cells before and after co-culture by stable isotope labeling by amino acids in cell culture (SILAC) method. We have shown that in co-cultured KG1a,
The effect of basic fibroblast growth factor (b-FGF), one of the commonest angiogenic factors in various cancer types, on lymphocyte adhesion and transmigration across the endothelial cell monolayer was investigated using human umbilical vein-derived endothelial cells (HUVEC) and type I collagen gel. Forty-eight h exposure of HUVEC with 2 ng/ml b-FGF significantly decreased the basal adhesion of lymphocytes to endothelial cells. The decrease ratio is further enhanced by the addition of shear stress in this assay system. When HUVEC was stimulated for the last 24 h with optimal conditions of recombinant interleukin 1β, the percentages of transmigration as well as adhesion were also decreased significantly by the presence of b-FGF. The expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 was down-regulated by b-FGF exposure in both resting and activated conditions by recombinant interleukin 1β, supposedly the main reason for this phenomenon. The migrating cells ...
The Neonatal Cardiac Endothelial Cell Isolation Kit, rat has been developed for the two-step isolation of vital cardiac endothelial cells from neonatal rat hearts (P0-P3). Enriched cardiac endothelial cells are fully functional and can be used for various downstream applications. - Lëtzebuerg
Maspin is a serpin that has multiple effects on cell behavior, including inhibition of migration. How maspin mediates these diverse effects remains unclear, as it is devoid of protease inhibitory activity. We have previously shown that maspin rapidly inhibits the migration of vascular smooth muscle cells (VSMC), suggesting the involvement of direct interactions with cell surface proteins. Here, using immunofluorescence microscopy, we demonstrate that maspin binds specifically to the surface of VSMC in the dedifferentiated, but not the differentiated, phenotype. Ligand blotting of VSMC lysates revealed the presence of several maspin-binding proteins, with a protein of 150 kDa differentially expressed between the two VSMC phenotypes. Western blotting suggested that this protein was the beta1 integrin subunit, and subsequently both alpha3beta1 and alpha5beta1, but not alphavbeta3, were shown to associate with maspin by coimmunoprecipitation. Specific binding of these integrins was also observed ...
Background: Recent studies have provided evidence that integrins play roles in recognition of mechanical stimuli and its translation into a cellular response. Integrin signaling may be regulated by a number of mechanisms including accessory proteins such as CD98 (4F2 antigen). Objectives: To determine CD98 expression by human articular chondrocytes and its involvement in human articular mechanotransduction. Methods: CD98 expression was assessed by immunostaining of cryostat sections of snap frozen articular cartilage and in cultured cells by western blotting. Chondrocytes enzymatically isolated from macroscopically normal and osteoarthritic (OA) articular cartilage were grown in short term, primary monolayer culture and used in a resting state or following mechanical stimulation at 0.33Hz. Results: Human articular chondrocytes express CD98 and immunoreactivity revealed a similar heterogeneous pattern of CD98 in both normal and osteoarthritic (OA) human articular cartilage. No role of CD98 was detected
Enolase-phosphatase 1 (ENOPH1), a newly discovered enzyme of the methionine salvage pathway, is emerging as an important molecule regulating stress responses. In this study, we investigated the role of ENOPH1 in blood brain barrier (BBB) injury under ischemic conditions. Focal cerebral ischemia induced ENOPH1 mRNA and protein expression in ischemic hemispheric microvessels in rats. Exposure of cultured brain microvascular endothelial cells (bEND3 cells) to oxygen-glucose deprivation (OGD) also induced ENOPH1 upregulation, which was accompanied by increased cell death and apoptosis reflected by increased 3-(4, 5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide formation, lactate dehydrogenase release and TUNEL staining. Knockdown of ENOPH1 expression with siRNA or overexpressing ENOPH1 with CRISPR-activated plasmids attenuated or potentiated OGD-induced endothelial cell death, respectively. Moreover, ENOPH1 knockdown or overexpression resulted in a significant reduction or augmentation of ROS
The results of this present thesis show a deficiency of IL-10 production in alveolar macrophages in asthma. The reduced IL-10 expression on protein and m-RNA level correlated with an increased production of pro-inflammatory cytokines such as TNF-(, MIP1- ( and GM-CSF. These observations implicate an impaired IL-10 synthesis in asthma with a subsequent prolongation of the inflammatory response. This leads to the conclusion that a dysbalance between pro- and anti-inflammatory cytokines is present in asthma and may be therefore of pathogenetic importance. The reduced sensitivity of alveolar macrophages to the inhibitory effects of exogenous IL-10 compared to peripheral blood monocytes may be caused by different signal transduction mechanisms. The expression of the proinflammatory cytokines RANTES and IL-8 in cultured human airway smooth muscle cells led to the conclusion that airway smooth muscle cells may act beside their contractile function as immunomodulatory cells in the pathogenesis of ...
Pattie Mathieu, Paul Cahill, Joseph Mackle, James King, Caitriona Lally, Phenotypic Changes in Rat Smooth Muscle Cells Exposed to Varying Amplitudes of Cyclic Equibiaxial Tensile Strain, UK Society of Biomaterials, Belfast, Ireland, 25th-26th June 2015 ...
AbstractIn the present work we set out to investigate the neuroprotective effects of noscapine (0.5-2 µM) in presence of D-glucose on primary murine foetal cortical neurons after oxygen-glucose deprivation/24 hrs recovery. Cell viability, nitric oxide production and intracellular calcium ([ca2+]i) levels were evaluated by MTT assay, the modified Griess method and Fura-2 respectively. 25 and 100 mM D-glucose could, in a concentration dependent manner, improve cell viability and decrease NO production and [ca2+]i level in neuronal cells after ischemic insult. Moreover, pre-incubation of cells with noscapine, noticeably enhanced protective effects of 25 and 100 mM D-glucose compared to similar conditions without noscapine pre-treatment. In fact, noscapine attenuated NO production in a dose-dependent fashion, after 30 minutes (min) OGD, during high-glucose (HG) condition in cortical neurons. Pretreatment with 2 μM noscapine and 25 or 100 mM D-glucose, was shown to decrease the rise in [ca2+]i induced by
Administration of selected concentrations of ebselen and N-acetyl cysteine have been proven to display an antioxidant potential based on their effect on markers of T cell integrity and function in human peripheral blood mononuclear cells and CD4+ T cell clones. Here we assessed the impact of various antioxidant concentrations on replicative aging of primary human fibroblast strains derived from embryonic lung (MRC-5) and foreskin (HFF). None of the antioxidant concentrations affected the cumulative population doublings, levels of oxidative DNA damage, intracellular GSH:GSSG ratio, potency of heat shock responses and the induction of senescence in both fibroblast strains. Our results showed no effect of both antioxidants on primary fibroblast strains and reveal their cell type specific antioxidant potential.
The effect of serum deprivation on proliferating cells is well known, in contrast its role on primary cell cultures, at confluence, has not been deeply investigated. Therefore, in order to explore the response of quiescent cells to serum deprivation, ubiquitous mesenchymal cells, as normal human dermal fibroblasts, were grown, for 48 h after confluence, in the presence or absence of 10% FBS. Fibroblast behaviour (i.e. cell morphology, cell viability, ROS production and elastin synthesis) was evaluated morphologically and biochemically. Moreover, the protein profile was investigated by 2-DE and differentially expressed proteins were identified by MS. Serum withdrawal caused cell shrinkage but did not significantly modify the total cell number. ROS production, as evaluated by the dihydroethidium (DH2) probe, was increased after serum deprivation, whereas elastin synthesis, measured by a colorimetric method, was markedly reduced in the absence of serum. By proteome analysis, 41 proteins appeared to ...
Haemopoietic stem cells in vivo proliferate and develop in association with stromal cells of the bone marrow. Proliferation and differentiation of haemopoietic stem cells also occurs in vitro, either in association with stromal cells or in response to soluble growth factors. Many of the growth factors that promote growth and development of haemopoietic cells in vitro have now been molecularly cloned and purified to homogeneity and various techniques have been described that allow enrichment (to near homogeneity) of multipotential stem cells. This in turn, has facilitated studies at the mechanistic level regarding the role of such growth factors in self-renewal and differentiation of stem cells and their relevance in stromal-cell mediated haemopoiesis. Our studies have shown that at least some multipotential cells express receptors for most, if not all, of the haemopoietic cell growth factors already characterized and that to elicit a response, several growth factors often need to be present at ...
A large selection of MEM formulations. Minimum Essential Medium (MEM), developed by Harry Eagle, is one of the most widely used of all synthetic cell culture media.
Several studies support C-reactive protein (CRP) as a systemic cardiovascular risk factor. The recent detection of CRP in arterial intima suggests a dual activity in atherosclerosis as a circulating and tissue mediator on vascular and immune cells. In the present paper, we focused on the inflammatory effects of CRP on human monocytes, which were isolated by Ficoll-Percoll gradients and cultured in adherence to polystyrene, endothelial cell monolayer, or in suspension. Chemokine levels, adhesion molecule, and chemokine receptor expression were detected by ELISA, flow cytometry, and real-time RT-PCR. Migration assays were performed in a Boyden chamber. Stimulation with CRP induced release of CCL2, CCL3, and CCL4 in adherent monocytes through the binding to CD32a, CD32b, and CD64, whereas no effect was observed in suspension culture. This was associated with CRP-induced up-regulation of adhesion molecules membrane-activated complex 1 (Mac-1) and ICAM-1 on adherent monocytes. Blockade of Mac-1/ICAM-1
Macrophages are mononuclear phagocytes derived from haematopoietic progenitors that are widely distributed throughout the body. These cells participate in both innate and adaptive immune responses and lie central to the processes of inflammation, development, and homeostasis. Macrophage physiology varies depending on the environment in which they reside and they exhibit rapid functional adaption in response to external stimuli. To study macrophages in vitro, cells are typically cultured ex vivo from the peritoneum or alveoli, or differentiated from myeloid bone marrow progenitor cells to form bone marrow-derived macrophages (BMDMs). BMDMs represent an efficient and cost-effective means of studying macrophage biology. However, the inherent sensitivity of macrophages to biochemical stimuli (such as cytokines, metabolic intermediates, and RNS/ROS) makes it imperative to control experimental conditions rigorously. Therefore, the aim of this study was to establish an optimised and standardised method for the
Growth and differentiation of osteoblasts are often studied in cell cultures. In vivo, however, osteoblasts are embedded within a complex three-dimensional (3D) microenvironment, which bears little relation to standard culture flasks. Our study characterizes osteoblast-like cells cultured in 3D collagen gels and compares them with cells in two-dimensional (2D) cultures. Primary rat osteoblasts and MC3T3-E1 cells were seeded within type I collagen gels, and differentiation was determined by mineral staining and gene expression analysis. Cells growing in 3D gels showed positive mineral staining and induction of osteoblast marker genes earlier than cells growing in 2D. A number of genes, including osteocalcin, bone sialoprotein, alkaline phosphatase and dentin matrix protein 1, were already highly upregulated in 3D cultures 24 h after seeding. The early expression of osteoblast genes was dependent on the 3D structure and was not induced in cells growing on collagen-coated dishes in 2D. Comparison of