Adult Progenitor Cells From Cord Blood Pluripotent Biological Characteristics And Separation Studies Of Related Factors

The present study was designed to investigate the affecting factors related to the isolation of multipotent adult progenitor cells (MAPCs) from human umbilical cord blood in low serum (2%) condition, optimize the isolation conditions and improve the yield of MAPCs. Using the optimal isolation conditions, we explored biological characteristics of MAPCs derived from umbilical cord blood.In PART Ⅰ, the affecting factors related to the isolation of MAPCs from human umbilical cord blood in low serum (2%) condition were investigate, The low-density mononuclear cells (MNC) from UCB of normal full term deliveries were isolated by Ficoll-Hypaque and cutured with diverse medium including 2% fetal bovine serum, and the effects of medium component, medium exchange time and initial plating density on isolation of MAPCs were studied. Result: The medium of DMEM/F12 was better than IMDM and DMEM—LG for MAPCs culture in low serum condition. The optimal yield of MAPCs was obtained when mononuclear cells were cultured at a initial plating density of 1 × 10~6 cells / cm~2 and the medium was exchanged to remove the nonadherent cells after 72 hours of inoculation. MAPCs isolated and cultured under the above conditions maintained a homogenous morphology, high potential ability of expansion and differentiation.In PART Ⅱ, the biological characteristics of MAPCs derived from umbilical cord blood and the mRNA expression of some cytokines promoting hematopoiesis on these cells were investigated. MAPCs from human full-term umbilical cord blood samples were isolated. The morphology, proliferation curve, cell cycle, immunophenotype, Osteogenic, Adipogenic and Neurogenic differentiation and the mRNA expression of some cytokines were assessed. MAPCs coud be isolated and cutured from 73 %(8/11) UCB samples. These cells displayed fibroblast-like morphology. Cell cycle analysis revealed that more than 90% of MSC cells were in G0/G1 phase. They were passaged over 8-10 times with no significant structural changes found. Flowcytometry revealed that MAPCs were negative for hematopoietic marker CD34, CD45, CD14, CD3, HLA-DR, endothelial cells marker CD31 and integrin CD11a, UCB-derived MAPCs...