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  • murine
  • Furthermore, disease models can be studied in a syngeneic human system circumventing the use of murine macrophages. (jove.com)
  • Mouse models studying macrophage biology suffer from the phenotypic and functional differences between murine and human monocyte-derived macrophages. (jove.com)
  • The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer were specifically developed to transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells.This video demonstrates how to establish primary hematopoietic cell cultures from murine bone marrow, and then prepare them for electroporation in the MXcell system. (jove.com)
  • vitro
  • The protocol presented here provides a simple and efficient way to deliver BMP-2 for in vitro studies on cells. (jove.com)
  • Both short- and long-term signaling events triggered by BMP-2 are taking place when cells are exposed to surfaces presenting covalently immobilized rhBMP-2, making this approach suitable for in vitro studies on cell responses to BMP-2 stimulation. (jove.com)
  • In vitro CSR assay involves the flow cytometric measurement surface Ig expression on activated B cells. (jove.com)
  • We have proposed that ubiquilin-1 prevents inappropriate inter- and intramolecular interactions of AICD, preventing aggregation in vitro and in intact cells 4 . (jove.com)
  • The protocol will promote research of TEC function and as well as the development of in vitro T cell reconstitution. (jove.com)
  • Therefore, there is an urgent need to establish proper in vitro and in vivo culture systems to study the action of novel therapeutics on human MM cells. (jove.com)
  • Here we present a model to grow human multiple myeloma cells in a complex 3D environment in vitro and in vivo . (jove.com)
  • We establish sensitive ELISA and WB assays for sCD4 detection in conditioned medium of in vitro cell culture system and serum of chronic inflammatory patients. (jove.com)
  • In vitro cell experiments are used to specifically evaluate cell type-dependent changes caused by a substance of interest, but culture-dependent variations and the inability to analyze the influence of specific molecules in the context of the inflammatory compound in atherosclerotic lesions limit the impact of the results. (jove.com)
  • This in vitro model is reliable for studying the biological activities of M1 and M2 macrophages and also for a proteomic approach. (jove.com)
  • Therefore, we here describe an in vitro model to generate and study primary human macrophages. (jove.com)
  • Lentiviral vectors have been successfully used in transducing many neural cell types in vitro 4-6 and in animals 7-10 . (jove.com)
  • The results from this study demonstrated that mesenchymal cells treated with peptide in vitro differentiate towards osteogenesis, and, that peptides delivered in vivo using a collagen sponge promote the repair of unicortical defects. (jove.com)
  • colony stimul
  • Interleukin-34 (IL-34) is a recently discovered cytokine functionally overlapping macrophage colony stimulating factor (M-CSF), a mediator of inflammation and osteoclastogenesis in bone-degenerative diseases such as rheumatoid arthritis. (jove.com)
  • These monocytes are then cultured in fluorinated ethylene propylene (FEP) Teflon-coated cell culture bags in the presence of macrophage colony-stimulating factor (M-CSF). (jove.com)
  • apoptosis
  • Immunohistochemical analyses showed that TC11 induced apoptosis of MM cells in vivo. (jove.com)
  • Excessive ROS, however, can exert cytotoxic effects, cause oxidative damage to proteins, and DNA, can interfere with cell growth and cell cycle progression, and induce apoptosis of gingival fibroblasts. (frontiersin.org)
  • Alhetheel A, Yakubtsov Y, Abdkader K, Sant N, Diaz-Mitoma F, Kumar A, Kryworuchko M (2008) Amplification of the signal transducer and activator of transcription I signaling pathway and its association with apoptosis in monocytes from HIV-infected patients. (springer.com)
  • receptors
  • Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty acid receptors housed on taste cells. (jove.com)
  • Various chemomkines and receptors are involved in the homing of hematopoietic stem cells. (jove.com)
  • With progesterone receptors predominantly localized on cells other than macrophages, the findings suggest that interactions between cells may facilitate the loss of progesterone receptor-mediated actions as part of a final common mechanism that remodels the cervix in certain etiologies of preterm and with parturition at term. (jove.com)
  • Moreover, in BMSC, wedelolactone enhanced binding of Sema3A with cell-surface receptors, including neuropilin (NRP)1 and plexinA1. (frontiersin.org)
  • The origin and migration pattern of DCs as well as their sharing of hematopoietic lineages, functions, and receptors with other phagocytic cell types such as monocytes and macrophages is a subject of intense investigation. (keystonesymposia.org)
  • Proteins
  • The M1 and M2 macrophages proteins are labelled with cyanine dyes before separation by isoelectric focusing, according to their isoelectric point in the first dimension, and their molecular mass, in the second dimension. (jove.com)
  • Lentiviral vectors are commonly produced by cotransfection of 293T cells with lentiviral transfer plasmid and the helper plasmids encoding the proteins required for vector packaging. (jove.com)
  • osteoblasts
  • After isolation of osteoblasts from mouse calvaria, all-trans retinoic acid (ATRA) is added to cell medium, and cell monitoring is conducted daily under an inverted microscope. (jove.com)
  • RAW264.7
  • Three clonal Ads knockdown RAW264.7 (RAW) macrophage cell lines with varying degrees of Ads expression and OCG deficiency were generated. (jove.com)
  • In this study, we found that wedelolactone, a compound isolated from Ecliptae herba, and a 9-day incubation fraction of conditioned media obtained from wedelolactone-treated bone marrow mesenchymal stem cell (BMSC) significantly inhibited tartrate-resistant acid phosphatase (TRAP) activity in RANKL-stimulated osteoclastic RAW264.7 cells. (frontiersin.org)
  • Addition of the semaphorin 3A (Sema3A) antibody to the conditioned media partially blocked the medium's inhibitory effects on the RAW264.7 cells. (frontiersin.org)
  • Blocking Sema3A activity with its antibody reversed wedelolactone-induced alkaline phosphatase activity in BMSC and concurrently enhanced wedelolactone-reduced TRAP activity in osteoclastic RAW264.7 cells. (frontiersin.org)
  • In osteoclastic RAW264.7 cells, conditioned media and wedelolactone promoted the formation of plexin A1-NRP1, but conditioned media also caused the sequestration of the plexin A1-DNAX-activating protein 12 (DAP12) complex and suppressed the phosphorylation of phospholipase C (PLC)γ2. (frontiersin.org)
  • In osteoclastic RAW264.7 cells, wedelolactone inhibited osteoclastogenesis through sequestration of the plexinA1-DAP12 complex, induced the formation of plexinA1-Nrp1 complex, and suppressed PLCγ2 activation. (frontiersin.org)
  • Therefore
  • Therefore, the most widely used cellular system is the osteocyte-like MLO-Y4 cell line. (jove.com)
  • The ability to identify in vivo macrophage activity in an infected joint can therefore provide two main applications: early detection of acute synovitis and monitoring of therapy. (jove.com)
  • Amyloid
  • peptides, analyzed in cell media from amyloid precursor protein (APP) transfected cells and in cerebrospinal fluid (CSF) from dogs by immunoprecipitation-mass spectrometry, using several different BACE1 inhibitors. (jove.com)
  • Amyloid precursor protein (APP) is a type I transmembrane protein associated with the pathogenesis of Alzheimer's disease (AD). (jove.com)
  • centrifugation
  • Thymic tissue is mechanically disrupted and after different steps of enzymatic digestion, the resulting cell suspension is enriched using a Percoll density centrifugation step. (jove.com)
  • describe
  • We describe here the use of a mouse embryonic stem (ES) cell based assay to identify critical time windows for Wnt/β-catenin and BMP signal activation during cardiogenic induction. (jove.com)
  • Here we describe a unique procedure for accurate measurement of IgE-producing mouse B cells that have undergone CSR in culture. (jove.com)
  • Because TECs are a rare cell population in the thymus, we also describe an effective way to enrich and purify TECs by depleting thymocytes, the most abundant cell type in the thymus. (jove.com)
  • In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and heterogeneity of patient-derived multiple myeloma (MM) cells. (jove.com)
  • As an example of this technique, we describe here the titration of glycogen in two cell lines, Chinese hamster lung fibroblast CCL39 and human colon carcinoma LS174, incubated in normoxia (21% O 2 ) versus hypoxia (1% O 2 ). (jove.com)
  • flow cytometry
  • The ability of transferred CDPs to contribute to the pDC population in PP and to respond to Flt3L was evaluated by flow cytometry of PP single cell suspensions from recipient mice. (jove.com)
  • Using the isolated cells, multiparameter flow cytometry can be applied to identification and characterization of TECs and dendritic cells. (jove.com)
  • After incubation for at least 72 hours and for up to five days at 37 °C, cells are subsequently analyzed for the capability to produce IL-17 through flow cytometry, qPCR, and ELISAs. (jove.com)
  • collagen
  • MM cell lines OPM-2 and RPMI-8226 were transfected to express the transgene GFP and were cultivated in the presence of human mesenchymal cells and collagen type-I matrix as three-dimensional spheroids. (jove.com)
  • Similar to the cervix before term, the preterm cervix of progesterone receptor antagonist-treated rats was characterized by reduced cell nuclei density, decreased collagen content and structure, as well as a greater presence of macrophages per unit area. (jove.com)
  • In contrast to the progesterone receptor antagonist-induced advance in characteristics associated with remodeling, ovariectomy-induced loss of systemic progesterone did not affect hypertrophy, extracellular collagen, or macrophage numbers in the cervix. (jove.com)
  • Profiling
  • Purified cells are suitable for various downstream analyses like Real Time-PCR, Western blot and gene expression profiling. (jove.com)
  • Interestingly, however, ligand occupancy does not appear to have a significant effect on HNF4alpha transcriptional activity, as evidenced by genome-wide expression profiling in cells derived from human colon. (jove.com)
  • cytoplasmic
  • A hidden and often overlooked resource of the macrophage rests in its high cytoplasmic nuclear ratios that allow the cell to sense its environment and rid it of the cellular waste products and microbial pathogens it encounters. (springer.com)
  • morphological
  • The method here described refers to the use of retinoic acid to generate a homogeneous population of ramified cells with morphological and molecular osteocyte features. (jove.com)
  • Secretion
  • Aging Disrupts Muscle Stem Cell Function by Impairing Matricellular WISP1 Secretion from Fibro-Adipogenic Progenitors. (nih.gov)
  • Stable secretion of high levels of hBDNF was detected in supernatants of transduced cells using western blot and ELISA. (jove.com)
  • hematopoietic
  • Homing is the phenomenon whereby transplanted hematopoietic cells are able to travel to and engraft or establish residence in the bone marrow. (jove.com)
  • 1, This paper outlines the classic homing protocol used in hematopoietic stem cell studies. (jove.com)
  • Enrichment of TEC populations (mTEC, cTEC) is achieved by depletion of hematopoietic (CD45 hi ) cells from the low-density Percoll cell fraction allowing their subsequent isolation via fluorescence activated cell sorting (FACS) using specific cell markers. (jove.com)
  • human
  • Investigating the Vascular Niche: Three-Dimensional Co-culture of Human Skeletal Muscle Stem Cells and Endothelial Cells. (nih.gov)
  • Cell sorting of various cell types from mouse and human skeletal muscle. (nih.gov)
  • Moreover, angiogenic responses of the host and invasion of tumor cells into the subjacent host tissue can be monitored daily by a stereo microscope and analyzed by immunohistochemical staining against human tumor cells (Ki-67, CD138, Vimentin) or host mural cells covering blood vessels (desmin/ASMA). (jove.com)
  • Moreover, similar findings are recapitulated in human primary CD4(+) T cells. (jove.com)
  • Human macrophages are involved in a plethora of pathologic processes ranging from infectious diseases to cancer. (jove.com)
  • The technique relies mostly on commonly available lab equipment and thus provides a cost and time effective way to obtain large quantities of human macrophages. (jove.com)
  • The goal of the two-dimensional (2D) electrophoresis protocol described here is to show how to analyse the phenotype of human cultured macrophages. (jove.com)
  • The orphan GPCR is transiently expressed in human embryonic kidney 293T (HEK293T) cells and a promiscuous Gα 16 construct is co-transfected. (jove.com)
  • Until now, neutralizing capacity of GM-CSF autoantibodies has been assessed by evaluating the growth inhibition of human bone marrow cells or TF-1 cells stimulated with GM-CSF 6-8 . (jove.com)
  • GM-CSF binding to sGMRα was dose-dependently inhibited by polyclonal GM-CSF autoantibody in a similar manner to the bioassay using TF-1 cells, indicating that our new cell-free assay system using sGMRα is more useful for the measurement of neutralizing activity of GM-CSF autoantibodies than the bioassay system using TF-1 cell or human bone marrow cells. (jove.com)
  • neuronal cell
  • High titer vectors were prepared for efficient transduction of neuronal cell lines, hMDM, and mMDM. (jove.com)
  • causes synaptic dysfunction and neuronal cell death, which are involved in the pathogenesis of Alzheimer's disease (AD). (jove.com)
  • mammalian cells
  • Here we utilize an affinity isolation/mass-spectrometry (AIMS) approach to demonstrate that HNF4alpha is selectively occupied by linoleic acid (LA, C18:2omega6) in mammalian cells and in the liver of fed mice. (jove.com)
  • absence
  • Digital image analysis software detects live and dead cells from presence or absence of membrane motion, and generates curves of change in viability as a function of drug concentration and exposure time. (jove.com)
  • Low-level viral persistence continues in these cells in the absence of macrophage death. (springer.com)
  • density
  • Lrp4 is a multifunctional member of the low density lipoprotein-receptor gene family and a modulator of extracellular cell signaling pathways in development. (jove.com)
  • To identify miRNAs regulated by estrogen signaling in breast cancer, their expression in ERα-positive breast cancer cell lines were compared before and after estrogen-activation using both the µParaflo-microfluidic microarrays and Dual Labeled Probes-low density arrays. (jove.com)
  • detection
  • The layout, including the specific conditions for cell culture and estrogen treatment, biological and technical replicates, and large-scale screening followed by in-depth confirmations using separate techniques, ensures a robust detection of miRNA regulations, and eliminates false positives and other artifacts. (jove.com)
  • primary
  • isolation of primary osteocytes is difficult and produces low cell numbers. (jove.com)
  • Daily cell monitoring is needed because of the inherent variability of primary cells, and the protocol can be adapted with minimal variation to cells obtained from different mouse strains and applied to transgenic models. (jove.com)
  • However, mutated or unknown miRNAs, or regulations at the primary and precursor transcript level, will not be detected. (jove.com)
  • It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. (jove.com)
  • RANKL
  • Gain of function mice overexpressed RANKL in T cells, whereas loss of function animals lacked expression of Traf6 in TECs (Traf6?TEC). (jove.com)
  • investigation
  • The proteomic approaches described here allows the investigation of the macrophage protein changes associated with various disorders like host pathogenicity or microbial toxins. (jove.com)
  • procedure
  • To further understand the biology of these cell types, characterization of these cell populations is highly desirable but due to their low frequency, isolation of any of these cell types requires an efficient and reproducible procedure. (jove.com)
  • endothelial
  • We demonstrate that 1) E2 regulates PDGF-BB production by endothelial cells in a dose-dependent manner and 2) optimization of PDGF-BB levels and induction of robust PDGF-mediated endothelial cell-vascular pericyte interactions require high (estrous) E2 concentrations. (jove.com)
  • We conclude that high (estrous) levels of E2 are important in controlling PDGF-mediated crosstalk between endothelial cells and pericytes, a fundamental mechanism governing microvessel stability and essential for preserving intracranial homeostasis. (jove.com)
  • Among the numerous BBB models thus far described, a static (without flow), contact BBB model, where astrocytes and brain endothelial cells (BECs) are cocultured on the opposite sides of a porous membrane, emerged as a simplified yet authentic system to simulate the BBB with high throughput screening capacity. (jove.com)
  • Receptor
  • Thus, loss of nuclear progesterone receptor-mediated actions promoted structural remodeling of the cervix, increased census of resident macrophages, and preterm birth much like that found in the cervix at term. (jove.com)
  • Prior to ligand screening, the receptor-expressing cells are loaded with a fluorescent calcium indicator, Fluo-4 acetoxymethyl. (jove.com)
  • The fluorescent signal of Fluo-4 is negligible in cells under resting conditions, but can be amplified more than a 100-fold upon the interaction with calcium ions that are released after receptor activation. (jove.com)
  • The HEK293T cells are easy to handle and have proven their efficacy throughout the years in receptor deorphanization assays. (jove.com)
  • OBJECTIVE: To mimic GM-CSF binding to GM-CSF receptor on the cell surface using cell-free receptor-binding-assay. (jove.com)
  • activity
  • These transduced cells were tested for hBDNF expression, stability, and neuroprotective activity. (jove.com)
  • dendritic
  • In some mice, overexpression of the dendritic cell growth factor Flt3 ligand (Flt3L) was enforced prior to adoptive transfer of CDPs, using hydrodynamic gene transfer (HGT) of Flt3L-encoding plasmid. (jove.com)
  • thus
  • Thus, the structure and macrophage census in the cervix appear sufficient for premature ripening and birth to occur well before term. (jove.com)
  • dyes
  • Fluorescence microscopy of cells loaded with fluorescent, Ca 2+ -sensitive dyes is used for measurement of spatial and temporal aspects of Ca 2+ signaling in live cells. (jove.com)
  • We use visible wavelength dyes to minimize photo-damage to the cells, but this means that ratiometric recording is not possible. (jove.com)
  • interactions
  • The close proximity of these cells naturally suggests that they interact with one another, but these interactions are just beginning to be characterized. (jove.com)
  • membrane
  • In the pancreas, highly intense Zip14 immunostaining was localized on perinuclear regions and the plasma membrane in acinar cells. (ufl.edu)
  • This carbohydrate epitope is carried by both glycolipids and glycoproteins expressed on the cell membrane. (beckman.com)
  • Purification
  • Following the enrichment, cell sorting time can be decreased so that loss of cell viability can be minimized during purification of TECs. (jove.com)
  • gene expression
  • Studies in chick and mouse demonstrate that secreted factors in the eCSF, fluid pressure, or a combination of these, are important for neurogenesis, gene expression, cell proliferation, and cell survival in the neuroepithelium 10-20 . (jove.com)
  • assay
  • RESULTS: The cell-free assay system using sGMRα yielded the data with high plasticity and reliability. (jove.com)
  • CONCLUSIONS: We established a cell-free assay quantifying the neutralizing capacity of GM-CSF autoantibody. (jove.com)
  • lines
  • The described technique does not require the time-consuming establishment of stably transfected cell lines in which the transfected genetic material is integrated into the host cell genome. (jove.com)
  • straightforward
  • While measurement of IgA and IgG subclasses is straightforward, measurement of IgE by this method is problematic due to soluble IgE binding to FcεRII/CD23 expressed on the surface of activated B cells. (jove.com)
  • mechanisms
  • Being in essence transcriptionally and translationally silent, these motile cells rely profoundly on diverse signaling mechanisms to orient themselves and swim in a directed fashion, and to contend with challenging environmental conditions during their journey to find the egg. (jove.com)
  • These phagocytic and intracellular killing sensing mechanisms can also be used in service as macrophages serve as cellular carriage depots for antiretroviral nanoparticles and are able to deliver medicines to infectious disease sites with improved therapeutic outcomes. (springer.com)