| Objective: To explore characteristics of human umbilical cord mesenchymal stemcells (hUC-MSCs) before and after cryopreservation supporting capacities in ex vivoexpansion of human hematopoietic stem/progenitor cells (HSPCs). Methods:Mesenchymal cells were isolated from umbilical cord cultured and expanded to P3and P6,cryopreservated for one and three months seperately, At the Third passage, cellular surfaceantigens were examined using flow cytometry, and induced to differentiate into osteoblastsand lipocytes. To explore hUC-MSCs supporting capacities in ex vivo expansion ofhuman HSPCs, experiment was divided into five groups: group A: Bone marrow nuclearcells (BMNCs); group B: BMNCs and fibroblast; group C: BMNCs and bone marrowmesenchymal stem cells; group D: BMNCs and umbilical cord mesenchymal stem cells. E:BMNCs umbilical cord mesenchymal stem cells with cryopreservation for one month.Proliferation and colony forming ability of hematogenesis and Stroma derived IL-3, IL-6,IL-11, SCF and TPO secretion levels were measured by Methylcellulose at0day,14day,28day,35day. Results:1. After passage1(P1) hUC-MSCs were cultured to P3andP6, the survival rate of non-cryopreserved cells was98.8%and96.3%and95.5%ofhUC-MSCs were survived after the cells were cryopreserved for1month and3months,respectively. The number of the original cells deceided the Growth characteristics.2.hUCMSCs with cryopreser-vation can differentiate into osteoblasts and adipocytes undercertain conditions.3. The colony-forming ability of hematogenesis expressing in differenttime of group Cã€D and E was significantly higher than those of group A and B（p＜0.05）.4. Group C and D still have colony forming of HSPCs in vitro long-term training35days, CD34+cells ratio increased after cultivate hematopoietic. Conclusion: hUC-MSCs before and after freezing can effectively support hematopoietic stem/progenitor cellsexpansion in vitro, especially maintain the earlier stem/progenitor cells expansion. |
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