cell-cell contact in transwell systems - posted in Tissue and Cell Culture: I would like to conduct transwell experiments with two cell types and wants to investigate the effect of cell-cell interaction between these 2 cell types. Do anyone have protocol of doing co-culture of these cells in the transwell system (ie. cell type 1 on the upper surface of the insert; while cell type 2 on the lower surface of the insert). Thank you very much.
Muscle regeneration involves the coordination of myogenesis and revascularization to restore proper muscle function. Myogenesis is driven by resident stem cells termed satellite cells (SC), whereas angiogenesis arises from endothelial cells and perivascular cells of preexisting vascular segments and the collateral vasculature. Communication between myogenic and angiogenic cells seems plausible, especially given the number of growth factors produced by SC. To characterize these interactions, we developed an in vitro coculture model composed of rat skeletal muscle SC and microvascular fragments (MVF). In this system, isolated epididymal MVF suspended in collagen gel are cultured over a rat SC monolayer culture. In the presence of SC, MVF exhibit greater indices of angiogenesis than MVF cultured alone. A positive dose-dependent effect of SC conditioned medium (CM) on MVF growth was observed, suggesting that SC secrete soluble-acting growth factor(s). Next, we specifically blocked VEGF action in SC ...
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 ...
TY - JOUR. T1 - Collagen-based co-culture for invasive study on cancer cells-fibroblasts interaction. AU - Che, Zhong Min. AU - Jung, Tae Heob. AU - Choi, Jung Hee. AU - Yoon, Do Jun. AU - Jeong, Hyun Jeong. AU - Lee, Eun Ju. AU - Kim, Jin. PY - 2006/7/21. Y1 - 2006/7/21. N2 - The roles of tumor stroma in carcinogenesis are still unclear. This study was aimed at designing an in vitro model for investigating the effects of stromal fibroblasts in the invasive growth of squamous cell carcinoma. Using two cancer cell lines, we performed three-dimensional co-culture with dermal equivalents to evaluate the effects of fibroblasts in cancer invasion. In vitro models for cellular interaction study were designed as follows: a collagen gel-based direct co-culture model (C-Dr) and a collagen gel-based indirect co-culture model (C-In). The invasive growth was found only in the dermal equivalents with fibroblasts. MMP-2 activity could be induced by direct contact between cancer cells and stromal fibroblasts. ...
Novel mechanisms, and molecular mediators, of the pro-tumorigenic effects of cancer-associated fibroblasts (CAFs) have been identified. These include CXCL12/SDF-1-mediated recruitment of bone marrow-derived endothelial precursor cell and pro-metastatic effects of CCL5. Co-culture experiments also su …
Formulations comprising combinations of APCs and tumor cells and APCs and virally infected cells are disclosed. These formulations generally comprise hybridoma of at least one antigen presenting cell
Cell culture inserts were seeded with 1x105 cells in 250 uL of medium with 0. 1% FBS. Un coated inserts had been utilised for migration assays whereas inserts
Trans-Lux Corp., a tmaker of digital LED and LCD display screens, cites Trumps election victory and his tough talk on manufacturing as the main reason it is bringing production back to the U.S.
Endothelial vascular smooth muscle cell coculture assay for high throughput screening assays to identify antiangiogenic and other therapeutic molecules George A TruskeyDepartment of Biomedical Engineering, Duke University, Durham, NC, USAAbstract: Drug development for many diseases would be aided greatly by accurate in vitro model systems that replicate key elements of in vivo physiology. The recent development of coculture systems of endothelial cells and vascular smooth muscle cells can be extended to high-throughput systems for the identification of compounds for angiogenesis, vascular repair, and hypertension. In this review, the various coculture systems are reviewed, and biologic interactions between endothelial cells and vascular smooth muscle cells are discussed. Key considerations in the design of high-throughput systems are presented, and selected examples are discussed.Keywords: endothelium, vascular smooth muscle cells, angiogenesis, microfluidics
In vitro coculture systems are at the forefront of molecular research due to their increased ability for cell-cell communication. In this study, small airway epithelial cells (SAEC) and human microvascular endothelial cells (HMVEC) were grown separately in monoculture or together in an alveolar-capillary coculture and exposed to either dispersion media control or multi-walled carbon nanotubes (MWC
The recent clinical successes of immune-based cancer therapies highlight the utility of harnessing the cytotoxicity of CD8 + T cells to target tumors. While current immunotherapies are effective at supporting and enhancing the function of tumor-specific T cell responses, these cells remain subject to multiple tolerizing mechanisms in the tumor microenvironment which can ultimately limit their effectiveness. While the function of dedicated regulatory immune cell populations like MDSCs and Tregs are well-studied contributors to tumor tolerance, our studies suggest that additional tumor-mediated mechanisms exist.. Our previous studies indicated that tumor cell lines secreted a soluble factor that was capable of inducing phenotypic and functional changes in normal human T cells that closely resembled those associated with dysfunctional T cells present in the periphery and tumor microenvironment of cancer patients [16, 17]. Because these studies were based on in vitro co-culture experiments, we first ...
BioTek Notas de Aplicación, 18-Feb-16, High Throughput, Multiplexed Detection of Inflammatory Cytokines in an Astrocyte and Monocyte Co-culture Model
Video articles in JoVE about salmonella typhimurium include Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization, Digital PCR-based Competitive Index for High-throughput Analysis of Fitness in Salmonella, Infection of Zebrafish Embryos with Intracellular Bacterial Pathogens, NF-κB-Dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells, Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria, In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration, Experimental Infection with Listeria monocytogenes as a Model for Studying Host Interferon-γ Responses, Functional Complementation Analysis (FCA): A Laboratory Exercise Designed and Implemented to Supplement the Teaching of Biochemical Pathways, Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers, Coincubation Assay for Quantifying
BACKGROUND. Prostate cancer (PCa) and bone cell interactions are critical in the metastatic phase. Kallikrein 4 (KLK4/hK4) is expressed in both PCa and mineralized tissues. We determined if KLK4/hK4 expression was associated with, and influenced by, the bone environment of metastatic PCa. METHODS. Immunohistochemistry, in vitro co-culture, cell migration, and attachment assays. RESULTS. hK4 was localized to tumor cells and osteoblasts in bone metastases. KLK4/hK4 increased in LNCaP and PC3 cells co-cultured with SaOs2 cells; SaOs2 KLK4/hK4 was unchanged. Co-culture did not affect cell proliferation but altered alkaline phosphatase activity/mRNA levels in SaOs2 cells. KLK4-transfected PC3 cells had increased migration towards SaOs2 conditioned medium and greater attachment to the bone-matrix proteins, collagens I and IV. CONCLUSIONS. hK4 expression and interaction with both tumor cells and osteoblasts suggests a role for hK4 in PCa bone metastasis. Whether this observation is unique to bone ...
Background P. gingivalis, a non-motile, rod-shaped, anaerobic, Gram-negative bacterium is one of the principal sources of periodontal disease. It possesses a number of potential virulence factors thought to be important in the disease process including 5 hemagglutinins (Hag). One of these is HagB. It is a well characterized nonfimbrial adhesin expressed on the surface of P. gingivalis. HagB is very pro-inflammatory and induces robust chemokine and cytokine responses in vitro and in vivo. Since the chemokine and cytokine responses seen from single cells grown in tissue culture often are not representative of the chemokine and cytokine profiles seen in clinical samples or biopsy specimens, we devised a co-culture model of keratinocytes, dendritic cells, and T-cells to test the hypothesis that chemokine and cytokine responses from co-cultured cells would be more representative of responses seen in clinical samples from individuals with periodontal disease than single cell models. Methods and materials HagB
Abstract We proposed an effective strategy for evaluating the targeting specificity of an antibody-conjugated quantum dot (QD) nanoprobe in a coculture system mimicking an in vivo-..
Our present study has provided, for the first time to our knowledge, a systematic analysis of the inflammation-relevant gene expression in ECs and SMCs in their coculture under static condition and in response to shear stress. We demonstrated that SMCs cause an upregulation of proinflammatory gene expression in ECs located in close proximity and that shear stress acts as a negative regulator for these proinflammatory gene expression in ECs cocultured with SMCs. Under static coculture condition, DNA microarray technology identified 23 inflammation-relevant genes that exhibit significant changes (18 increases and 5 decreases) in their expression in ECs cocultured with SMCs, as compared with the control monocultured ECs. All 18 genes upregulated in ECs by the coculture encode products that serve proinflammatory and thrombogenic functions. These functions include: (1) mediation of leukocyte-EC interaction and signal transduction (ICAM-1, VCAM-1, CD44, and NK-4); (2) promotion of chemotaxis and ...
1. Using sterile forceps, pick up the coverslips containing neurons and place into the bottom of the co-culture plate. Be sure to keep the cells covered with medium. Replace the insert with the microglia coverslip. Alternatively, you could change the medium in the microglial culture completely, and do a wash with fresh medium, before replacing the medium, to get rid of the LPS. Then add the neurons ...
Lastly, we examined heterotypic interactions between tumor cells and cancer associated fibroblasts (CAFs) to understand the mechanism of resistance to lapatinib. Using a 3D co-culture model, we identified significant sub-type-specific changes in gene expression, metabolic, and therapeutic sensitivity profiles of breast cancer cells induced by CAFs. We identified JAK2/STAT3 pathway and CAF-secreted hyaluronan as major factors contributing to CAF-mediated protection. We also found that close spatial proximity to CAFs impacts therapeutic responses by affecting proliferation rates of cancer cells ...
In model 1, target CD4+ T cells (UC blood CD4+ T cells and a T cell line expressing CXCR4 and CD4.403) are cocultured with effector HEK cells with or without Env expression. In model 2, HEK/CD4.403/CXCR4 and HEK/CD4.403 cells are cocultured with effector T cells with or without Env expression. In model 3, target CD4+ T cells are cocultured with HIVNL4-3-infected or uninfected T cells. Expression of gp120 at the surface of HEK.Env, 8.E5, and HIVNL4-3-infected effector cells as well as parental HEK and CEM cells - used as negative controls - were analyzed by flow cytometry after incubation of the cells with PBS (white histograms) or with PBS containing anti-gp120 human polyclonal Ab (black histograms). Bound Ab was detected with a secondary FITC-labeled goat anti-human Ig. Expression analysis of the extracellular domains of CD4 and CXCR4 at the surface of target cells was performed after incubation of the cells with PBS (white histograms) or with anti-CD4 (black histograms) or anti-CXCR4 (gray ...
In the present study, we set out to determine the degrees of residual plasma viremia in a large number of infected individuals receiving ART and to determine immunological or virological parameters that may correlate with residual plasma viremia. Residual plasma viremia was determined in quadruplicate using an automated system to minimize quantitative errors often associated with the detection of extremely low levels of viral RNA. We have demonstrated detectable levels of residual plasma viremia (1-49 copies/mL) in the majority of study participants receiving ART in whom plasma viremia had been suppressed for extended periods of time, as measured by a standard clinical assay (with a limit of detection of 50 copies/mL). Furthermore, we found a correlation between the level of residual plasma viremia and the frequency of CD4+ T cells carrying HIV proviral DNA. Of note, due to the limited amounts of blood obtained from the study subjects, we could not conduct the quantitative coculture assays that ...
Molostvov G, Morris A, Rose P, Basu S, Muller G (February 2004). "The effects of selective cytokine inhibitory drugs (CC-10004 and CC-1088) on VEGF and IL-6 expression and apoptosis in myeloma and endothelial cell co-cultures". British Journal of Haematology. 124 (3): 366-75. doi:10.1046/j.1365-2141.2003.04777.x. PMID 14717786 ...
OBJECTIVE: Rheumatoid arthritis (RA) is classically thought of as a Th1, T lymphocyte-driven disease of the adaptive immune system. However, cells of the innate immune system, including neutrophils, are prevalent within the diseased joint, and accumulate in large numbers. This study was undertaken to determine whether cells of the rheumatoid stromal microenvironment could establish an inflammatory environment in which endothelial cells are conditioned in a disease-specific manner to support neutrophil recruitment. METHODS: Human umbilical vein endothelial cells (ECs) and fibroblasts isolated from the synovium or skin of RA patients were established in coculture on opposite sides of porous transwell filters. After 24 hours of EC conditioning, the membranes were incorporated into a parallel-plate, flow-based adhesion assay and levels of neutrophil adhesion to ECs were measured. RESULTS: ECs cocultured with synovial, but not skin, fibroblasts could recruit neutrophils in a manner that was dependent on the
IncuCyte® NeuroPrime Cell Kit offers a complete solution for kinetic, live-cell imaging of 96-well co-culture assays utilizing primary rat neurons & astrocytes.
Conjugation, the transfer of DNA by direct cell-to-cell contact, depends on the presence of a conjugative plasmid(is small, double-stranded DNA molecules
Hierarchical tissues composed of spheroid and fiber structures such as tumors, embryos and glomeruli widely exist in organisms. Methods have been developed to build spheroid and fiber structures, independently, as tissue models in vitro. However, it is still a challenge to print them simultaneously and integrated f
The underlying technique our lab is built upon is a hematopoetic/mesenchymal stem cell coculture. In this system we are able to acurately recreate the bone marrow niche in-vitro, manipulate the microenvironment, and investigate the effects of such manipulations on the niche using fluorescence imaging, flow cytometry, in situ immunohistochemistry, and serial transplants.. ...
High expression of B7-H3 on stromal cells defines tumor and stromal compartments in epithelial ovarian cancer and is associated with limited immune activation:
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Ultrastructure of control and GGF-treated cocultures. Schwann cell-DRG neuron cocultures that had myelinated for 3 wk were maintained for an additional 3 d with
Purpose: : Evidences from literature shows that GITR-GITRL interaction plays important role in delivering co-stimulatory signals controlling regulatory T cells. Human GITR ligand has recently been shown to be constitutively expressed in retinal tissues. By expressing eYFP tagged human GITRL in human retinal pigment epithelial (RPE) cells, we investigated the biological significance of expression of GITRL on human ocular tissue. Methods: : Full length GITRL was amplified from a human ovarian cDNA library using GITRL specific primers and subcloned in a pEYFPC1 vector for expressing GITRL as an eYFP tagged protein in RPE cells. Using an RPE/T cell co-culture system we studied the invitro proliferative responses of purified CD3+ T cells or CD3+CD25-T cells co-cultured with lethally irradiated RPE cells in the presence or absence of anti-CD3 and anti CD28 antibody. For cytokine measurement, culture supernatants were collected from parallel RPE/T cell co-culture experiments. Results: : Both ...
Epidemiological studies have linked obesity with basal-like breast cancer risk in both premenopausal and postmenopausal women, suggesting that obesity may affect tumor phenotype by skewing the microenvironment toward support of more aggressive tumor cell phenotypes (typically ER/PR−, HER-2−). In order to study the interactions between adipocytes and breast tumor cells, we developed an adipocyte-breast cancer cell co-culture system. The adipocytes SGBS (Simpson-Golabi-Behmel Syndrome) and human ER positive breast cancer MCF7 cells were co-cultured for 24 hours. Following co-culture, the cells from the inserts (MCF7 cells) and wells (SGBS cells) were collected separately and total RNA was then isolated. The mRNA levels of the NADPH oxidase subunit NOX4 and hypoxia-inducible factor 1α (HIF1α) were determined by RT-PCR and normalized to 18S RNA levels. When SGBS cells were co-cultured with MCF7 cells, SGBS NOX4 and HIF1α mRNA levels increased more than 10 fold and 3 fold, respectively, while ...
LY317615 (Enzastaurin) IC50 in tumors possess different metastasis potential. Although microfluidic technology provides better control over co-culture microenvironment, many systems insert hundreds or hundreds of cells in the gadget still, therefore they absence one cell quality as typical co-culture in petri meals.6C14 Although the solo cell co-culture on-chip allows for isolating solo cells in the step, there are even now two critical problems to be resolved: 1) Thanks to the small amount of secreted protein from solo cell, constant perfusion can easily wash apart the secretion and impair cell-cell interaction thus; and 2) As the system goals to research the heterogeneity of one cells, the chamber-chamber cross-talk, which can trigger unwanted connections, should end up being removed. In the prior functions reported on the one cell-cell connections15,16, the co-culture microenvironment of each cell group was not isolated completely. Hence, the cross-talk among different co-culture environments ...
Additional file 2: Figure S2. of A human macrophage - hepatocyte co-culture model for comparative studies of infection and replication of Francisella tularensis LVS strain and subspecies holarctica and mediasiatica
LY317615 (Enzastaurin) IC50 in tumors possess different metastasis potential. Although microfluidic technology provides better control over co-culture microenvironment, many systems insert hundreds or hundreds of cells in the gadget still, therefore they absence one cell quality as typical co-culture in petri meals.6C14 Although the solo cell co-culture on-chip allows for isolating solo cells in the step, there are even now two critical problems to be resolved: 1) Thanks to the small amount of secreted protein from solo cell, constant perfusion can easily wash apart the secretion and impair cell-cell interaction thus; and 2) As the system goals to research the heterogeneity of one cells, the chamber-chamber cross-talk, which can trigger unwanted connections, should end up being removed. In the prior functions reported on the one cell-cell connections15,16, the co-culture microenvironment of each cell group was not isolated completely. Hence, the cross-talk among different co-culture environments ...
Video articles in JoVE about retinoblastoma protein include In Vivo Detection and Analysis of Rb Protein SUMOylation in Human Cells, Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software, Analysis of Cell Cycle Position in Mammalian Cells, Induction of Protein Deletion Through In Utero Electroporation to Define Deficits in Neuronal Migration in Transgenic Models, Whole Mount Dissection and Immunofluorescence of the Adult Mouse Cochlea, In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration, An In Vitro Model for Studying Cellular Transformation by Kaposi Sarcoma Herpesvirus, A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis, Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets, Assessing Replication and Beta Cell Function in Adenovirally-transduced
Methods Human MSCs were transduced with a lentiviral vector containing TRAIL IRES-GFP under the control of a tetracycline dependent promoter. Successful transduction was measured using flow cytometry and immunoblotting. The biological activity of MSCTRAIL was determined using co-culture experiments. 5×105 MPM cells were stained withDiI and plated with 5×105 MSCTRAIL cells. After 24 h doxycycline (10 μg/ml) was added to induce TRAIL production and left for 48 h. Both cells and supernatant were collected and stained with Annexin V and DAPI to detect apoptosis and death respectively onflow cytometry. ...
Applying biological molecules from cell membranes to the surfaces of artificial materials is opening peepholes on the very basics of cell-to-cell interaction.
Glioblastoma (GBM) is a tumor of the central nervous system. After surgical removal and standard therapy, recurrence of tumors is observed within 6-9 months because of the high migratory behavior and the infiltrative growth of cells. Here, we investigated whether carnosine (β-alanine-l-histidine), which has an inhibitory effect on glioblastoma proliferation, may on the opposite promote invasion as proposed by the so-called
Bacterial mating or conjugation is the transfer of DNA from one bacterium to another via direct cell-to-cell contact through a mating pore. My current research uses the genetically-tractable bacterium Bacillus subtilis as a model system to explore the function and subcellular localization of a putative component of the bacterial mating pore apparatus. I have been characterizing the protein ConE (formerly YddE) which is encoded on the B. subtilis conjugal element ICEBs1. ConE is related to proteins encoded on conjugal elements in numerous bacteria, including the Gram-positive pathogens S. aureus, C. difficile, and L. monocytogenes. ConE belongs to a large superfamily of ATP-dependent pumps involved in the extrusion of proteins and DNA through membrane pores. I have shown that ConE and its ATPase domain are essential for mating of ICEBs1. In addition, ConE-GFP localizes at the cell poles, in close association with the membrane (see Figure). Given ConEs localization, ATPase domain, and ...
Bacterial mating or conjugation is the transfer of DNA from one bacterium to another via direct cell-to-cell contact through a mating pore. My current research uses the genetically-tractable bacterium Bacillus subtilis as a model system to explore the function and subcellular localization of a putative component of the bacterial mating pore apparatus. I have been characterizing the protein ConE (formerly YddE) which is encoded on the B. subtilis conjugal element ICEBs1. ConE is related to proteins encoded on conjugal elements in numerous bacteria, including the Gram-positive pathogens S. aureus, C. difficile, and L. monocytogenes. ConE belongs to a large superfamily of ATP-dependent pumps involved in the extrusion of proteins and DNA through membrane pores. I have shown that ConE and its ATPase domain are essential for mating of ICEBs1. In addition, ConE-GFP localizes at the cell poles, in close association with the membrane (see Figure). Given ConEs localization, ATPase domain, and ...
In the present work, we report evidence that in addition to its role on the proliferation of pancreatic progenitor cells, the mesenchyme is crucial in controlling the timing of pancreatic β-cell differentiation. When we cultured rat embryonic pancreatic epithelium in the absence of its surrounding mesenchyme, we found that Ngn3 expression was turned on rapidly and then turned off a few days later, in keeping with the reported in vivo pattern (15,25). Our filter-separation experiments indicate that the mesenchyme-induced delay in Ngn3 induction requires direct contact between the epithelium and the mesenchyme. This result fits in with recent data indicating that direct cell-cell contact between epithelial and mesenchymal cells suppresses β-cell formation (26). We also found that Ngn3 expression occurred far earlier without mesenchyme, within a few hours compared with 3 days with mesenchyme either in vitro or in vivo. This acceleration in Ngn3 induction resembles the pancreatic phenotype of mice ...
The present invention relates to a multicellular, three-dimensional, living mammalian tissue. The tissue is produced by a co-culture process wherein two distinct types of mammalian cells are co-cultured in a rotating bioreactor which is completely filled with culture media and cell attachment substrates. As the size of the tissue assemblies formed on the attachment substrates changes, the rotation of the bioreactor is adjusted accordingly.
The differential expression analysis revealed that the expression of 34 genes were significantly up-regulated in CAFs compared with those in NFs. Furthermore, the pathway analysis showed that cytoskeletal signaling pathway was up-regulated and TGFb1 played crucial roles as one of upstream regulators in CAFs. Real-time imaging showed that the motility of CAFs was significantly greater on extracellular matrix. Intriguingly, the motility of CAFs conferred invasiveness on GC cells co-cultured with CAFs. ...
HYBRID REFRIGERATED CENTRIFUGE All in one unit from micro tube rotors to multi-place swing rotors. A reliable trump card for centrifuge... read more ...
This study demonstrates that the Corning 384-well spheroid microplate can be used to generate and perform viability assays with co-culture spheroids in an easy-to-use, high throughput format.
When I wrote about the NRDCs new Stewardship Index for Specialty Crops, and asked whether industrial monoculture was the real path to sustainable farming, the response from many of our readers was unsurprisingly
The macrophage colony-stimulating factor-deficient bone marrow stromal cell line OP9, derived from osteopetrotic mice, is known to support hematopoietic stem cell (HSC) expansion as well as hematopoietic differentiation of embryonic stem cells. Coculture of HSC in the OP9 system requires cytokine support to achieve significant cell expansion. Recently, we reported extensive expansion without cell senescence of cord blood (CB)-derived HSC cocultured with OP9 stromal cells for more than 18 weeks with a single cytokine support using human thrombopoietin (TPO). In this study, we evaluated the efficiency of the OP9/TPO coculture system to sustain long-term hematopoiesis of adult, granulocyte colony-stimulating factor mobilized human peripheral blood (PB) CD34(+) cells. Maximum cell expansion was attained during the first 4 weeks of coculture. At the same time, the maximum progenitor cell expansion was demonstrated by the production of colony-forming cells and cobblestone area-forming cells. In contrast to
Obesity is associated with a state of chronic, low-grade inflammation. Recent studies have demonstrated that obese adipose tissue is characterized by increased infiltration of macrophages, suggesting that they are an important source of inflammation in the adipose tissue.13-15,24 It is, therefore, important to elucidate the signals that attract macrophages to the adipose tissue and to dissect the molecular mechanisms whereby adipocytes and macrophages communicate within the adipose tissue. Here we developed an in vitro coculture system composed of differentiated 3T3-L1 adipocytes and the macrophage cell line RAW264. The data herein suggest that our coculture system provides the unique in vitro experimental model system to investigate the functional interaction between adipocytes and macrophages within the adipose tissue.. This study demonstrates for the first time that the coculture of 3T3-L1 and RAW264 results in marked upregulation of proinflammatory cytokines, such as MCP-1, IL-6, and TNF-α, ...