ABSTRACT
<p><b>OBJECTIVE</b>To assess the differentiation potential of rat adipose-derived stem cells (ADSCs) into Schwann-like cells in vitro.</p><p><b>METHODS</b>ADSCs isolated from adult SD rats were cultured in vitro and identified with the cell surface antigens CD44, CD49d and CD106 by immunocytochemistry. The ADSCs of the sixth to eighth passages were inoculated in polylysine-coated culture plate and cultured for 12 days in DMEM/F12 culture medium containing 10% fetal bovine serum, 5 ng/ml platelet-derived growth factor, 10 ng/ml basic fibroblast growth factor, 14 micromol/L Forskolin and 200 ng/ml Heregulin to induce their differentiation in vitro. Immunocytochemistry was performed to identify the expression of the cell surface markers nestin, glial fibrillary acidic protein (GFAP), S-100, and P75.</p><p><b>RESULTS</b>The isolated and purified ADSCs were positive for CD44 and CD49d expressions but negative for CD106. After 12 days of culture in the conditional culture medium, most of the cells showed positive expressions of GFAP, S-100, and P75, the specific protein markers of Schwann cells.</p><p><b>CONCLUSION</b>Adult rat ADSCs are confirmed to have potentials of neuroglial differentiation and capable of differentiating into Schwann-like cells in vitro.</p>
Subject(s)
Animals , Cattle , Male , Rats , Adipose Tissue , Cell Biology , Cell Differentiation , Cell Proliferation , Cytological Techniques , Methods , Gene Expression Regulation , Rats, Sprague-Dawley , Schwann Cells , Cell Biology , Metabolism , Stem Cells , Cell BiologyABSTRACT
@#ObjectiveTo investigate the feasibility and optimal condition of isolation,purification and expansion of mesenchymal stem cells(MSCs) derived from human umbilical cord blood in vtro.MethodsHuman umbilical cord blood(HUCB) was collected from full term deliveries scheduled,all samples were obtained sterilely with 20 U/ml preservative free heparin.The cord mononuclear cells were isolated with lymphocyte separation medium(density 1.077 g/ml),purified and expanded with MesencultTM medium and acidic environment to produce adherent layer.The surface antigen expression of MSCs was detected with flow cytometry.ResultsThe HUCB-derived mononuclear cells,when seeded in specific medium,gave rise to adherent cells,which exhibited either an osteoclast or mesenchymal-like phenotype.After passage 3,these cells were able to be purified and expanded.6.6×105 primary MSCs reached a number of 9.9×10<>sup8 after 10 expanded passage.Flow cytometry showed that MSCs did not express antigens CD34,CD11a and CD11b,but express strongly CD29 and weakly CD71,which was identical to human bone marrow-derived MSCs.ConclusionMSCs in HUCB can be cultured and expanded in vitro,and could be a source of stem cells for experimental and clinical application.