人胎盘源间充质干细胞对脐血单个核细胞体外生长的支持作用

Support of Human Placenta-derived Mesenchymal Stem Cells on the Growth of Mononuclear Cells from Umbilical Cord Blood

目的探讨人胎盘源间充质干细胞(PMSCs)对脐血单个核细胞(MNCs)体外生长的支持作用。方法将人胎盘组织经胶原酶消化、贴壁培养获得人PMSCs,用流式细胞仪检测细胞表面标志。以密度梯度离心法分离脐血MNCs和传代后的PMSCs进行共培养,通过细胞计数和流式细胞仪分别检测细胞的生长情况。结果从人胎盘组织中成功分离和培养PMSCs,经流式细胞仪检测其表型为CD29+、CD44+、CD105+、CD106+、CD166+、CD34-、CD45-、HLA-DR-。人PMSCs+脐血MNCs共培养组的细胞总数和CD45+细胞数在各培养时间点均明显高于对照组,差异有统计学意义(P<0.05或P<0.01)。在培养第7天,CD45+、CD14+及CD19+细胞数共培养组也明显高于对照组,差异有统计学意义(P<0.05或P<0.01)。结论人PMSCs可以作为滋养层细胞有效支持脐血MNCs的体外生长。

Objective To explore the role of human placenta-derived mesenchymal stem cells （PMSCs） in supporting the growth of mononuclear cells （MNCs） from umbilical cord blood （CB）. Methods Human placenta was digested with collagenase and the adherent cells were obtained after cultured. The phenotype of PMSCs was detected by flow cytometry.CB-derived MNCs isolated using Ficoll density centrifugation were cocuhured with PMSCs as feeder layer. The growth characteristics were analyzed by cell count and flow cytometry. Results PMSCs were successfully isolated and cultured from human placenta. FACS analyses showed that the phenotypes of PMSCs were CD29^＋, CD44^＋, CD105^＋, CD106^＋, CD166^＋, CD34^-, CD45^-, HLA-DR^-. The total cell number and CD45^＋ cells in the PMSCs＋CB MNCs group at the various culture time points were significantly higher than that in the CB MNCs alone group （P〈0.05 or P〈0.01）. On day 7, the amount of CD45^＋, CD14^＋ and CD19^＋ cells in the coculture group was also significantly higher than that in the control group separately （P〈0.05 or P〈0.01）. Conclusion Human PMSCs can support the growth of CB MNCs in vitro effectively as feeder layer.