Experimental Research On Construction Of Small Diameter Tissue Engineering Blood Vessel With Bone Marrow Mesenchymal Stem Cells

Part I: Isolation and Identification of Human Bone MarrowMesenchymal Stem CellsObjective To harvest, isolate , culture and identify bone marrow mesenchymal stem cells(BMMSCs) from human ribs. BMMSCs were purified and the growth characteristic was observed in vitro so that to investigate the possibility of BMMSCs as seed cells for tissue engineering.Methods BMMSCs were harvested from human ribs and isolated, purified by methods of density gradient centrifugation and adhering to the culture plastic. Cells of P₂ to P₃ were examined with flow cytometry for the presence or absence of special hematopoietic and endothelial markers(CD34, CD45.CD1la , CD14, KDR, CD105, HLADR). Cell growth curve of P₀, P₅, P₁₀ passage culture of human BMMSCs was drawn according to cells counting.Results Methods of density gradient centrifugation and adhering to the culture plastic can harvest mononuclear cells with specific characterics. The cells showed presence of KDR, CD105 and absence of CD34, CD45, CDlla, CD14, HLADR. Growth curve showed that there was difference of latency, activity and flat periods between primary culture and passage culture.Conclusion 1. Methods of density gradient centrifugation and adhering to the culture plastic can harvest higher purified BMMSCs. 2. BMMSCs showed presence of KDR, CD105 and absence of CD34, CD45, CDlla, CD14, HLADR. 3. BMMSCs...