Isolation And Identification Of Mesenchymal Stem Cells From Human Umbilical Cord Blood In Vitro

【Objective】1. To investigate the feasibility and optimal condition of isolation and amplification of mesenchymal stem cells from human umbilical cord blood (HUCB) in Vitro.2. To investigate difference of human unbilical blood derived from mesenchymal stem cells in differential culture conditions.【Methods】Human umbilical cord blood was collected in asepsis condition, anticoagulanted by sodium citrate compound , isolated by density gradient centrifugation,,these cells were proliferated clonally in two specific medium respectively,dected surface signs of MSCs by flow cytometry. The cord mononuclear cells were isolated with lymphocyte separation medium (density 1.077 g/ml), purified and expanded with specific medium, the surface antigen expression of MSCs was detected with flow cytometry.【Results】1. The Human umebilical cord blood 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. Flow cytomery showed that MSCs did not express antigens CD14,CD19,but express strongly CD105,CD44. 2. Compared the number of the forming clones of high concentration of fetal calf serum mononuclesr cells with low concentration of fetal calf calf serum,there were significant differences(P<0.05).【Conclusion】1. MSCs in HUCB can be cultured and expanded in vitro and could lay the foundation for further study.2. 30% fetal calf serum concentration medium markedly improved the application of human umbilical cord blood MSCs culture success rate.