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目的探讨人骨髓间充质干细胞(hMSCs)体外造血分化潜能。方法选用孕125-145d(125-145dpc)的昆明小鼠,分别制备小鼠胎肝基质细胞条件培养液(FLSC-CM)及胚胎成纤维细胞饲养层(FD),将体外扩增的CD34-CD45-hMSCs分别接种于含FLSC-CM、FD和IL-6及SCF组合的培养体系中,培养7d后,通过形态学、表型、粒-单/巨噬细胞系集落培养(CFU-GM)对分化细胞进行鉴定。结果hMSCs与FLSC-CM共培养组产生的非贴壁细胞明显增多,形态类似于单核或小淋巴细胞,部分细胞可表达人造血细胞特异性表面分子(CD34和CD45),在含人粒-单集落刺激因子(GM-CSF)的甲基纤维素培养体系中能够形成CFU-GM,而FD和IL-6+SCF诱导组无上述作用。结论FLSC-CM可诱导CD34-CD45-hMSCs分化为造血细胞,提示hMSCs具有体外造血分化潜能。
Objective To investigate the in vitro hematopoietic differentiation potential of human bone marrow mesenchymal stem cells (hMSCs). Methods Kunming mice (125-145dpc) were enrolled in this study. Fetal liver fibroblast feeder cells (FLSC-CM) and mouse embryonic fibroblast feeder layer (FL) were prepared respectively. CD34- CD45-hMSCs were respectively inoculated into the culture system containing FLSC-CM, FD and IL-6 and SCF combination. After culturing for 7 days, CD45-hMSCs were cultured in vitro by morphological, phenotypic and CFU- Differentiated cells were identified. Results The number of non-adherent cells in hMSCs co-cultured with FLSC-CM was significantly higher than that in mononuclear or small lymphocytes. Some cells expressed human hematopoietic cell-specific surface molecules (CD34 and CD45) CFU-GM was able to form in the methylcellulose culture system of colony stimulating factor (GM-CSF), while the FD and IL-6 + SCF inducing groups did not. Conclusion FLSC-CM can induce the differentiation of CD34-CD45-hMSCs into hematopoietic cells, suggesting that hMSCs have the potential of hematopoietic differentiation in vitro.