Comparison of isolation ways of MSC and VEGF and SDF-1αconcentration in respective culture medium / 重庆医学

ABSTRACT

Objective To choose one protocol that can quickly ,safely and effectively provide amount enough of bone marrow derived mesenchymal stem cells(MSCs) for use of clinical or experimental test through comparison of their growth characteristics and growth factors levels in culture solution .Methods Cells extracted from bone marrow of C57BL/C mice respectively underwent two different isolation protocolswhole bone marrow adherent culture(WBMAC) or gradient density separation(GDS);characteris‐tic surface antigens of MSCs were identified by flow cytometry on cells isolated in different ways ;the distinct growth curve of pri‐mary stem cells cultured in vitro described their different proliferation rate;levels of vascular endothelial growth factor(VEGF) and stromal cell‐derived factor‐1α(SDF‐1α) in culture medium were detected by ELISA .Results Primary MSCs obtained by WBMAC proliferated at higher speed and exhibited shorter growth cycle than those separated by GDS ;on MSCs from both groups ,surface antigens CD29 were detected positively ,and antigens including CD31 ,CD34 and CD45 were assayed negatively ;concentration of VEGF and SDF‐1αin both two nutrient solution primarily keep at low levels ,comparatively ,level of VEGF and SDF‐1αin the di‐shes which contain MSCs by WBMAC was higher than the one in the dishes which contain MSCs by GDS .Conclusion MSCs ex‐tracted by WBMAC shows unimpaired cell function ,can build automatically more suitable microenviroment for their growth;this classic method was qualified for clinical and experimental use in a safe ,rapid ,effective way .