THE ISOLATION PURIFICATION AND CULTURE OF HUCA MSCs

Objective: To investigate the isolation, purification and culture of human umbilical cord blood(HUCB) mesenchymal stem cells(MSCs) in vitro, and to observe the feasibility of using MSCs as seed cells in experiment and clinic. Methods: HUCB were collected from full term deliveries scheduled for cesarean section, all specimens was obtained sterilely with preservative-free heparin, the cord blood mononuclear cell was isolated by lymphocyte separation medium, and purified and culture with Mesencult TM medium and acidic environment to produce adherent layer, the surface antigen expression of MSCs was detected by flow cytometry. Results: The UCB-derived mononuclear cells, when set in culture, gave rise to adherent cells, which exhibited either an osteoclast-or mesenchymal-like phenotype, cells with the MSCs displayed a fibroblast-like morphology and expressed several MSCs-related antigens (CD 29, CD 44, CD 166), but did not express haematopoietic cells antigens(CD 34, CD 45), which was identical to human bone marrow-derived MSCs. Conclusion: MSCs in HUCB can culture and expand in vitro, which could be regarded as an alternative source of MSCs for experimental and clinical needs.