人脐带和骨髓间充质干细胞体外分离培养及生物学特性比较

Comparison of biological characteristics of human umbilical cord mesenchymal stem cells with human bone marrow mesenchymal stem cells

目的体外分离培养、扩增人脐带间充质干细胞(h UC-MSC)和人骨髓间充质干细胞(h BM-MSC),并对其生物学特性进行比较。方法采用组织块贴壁法从足月胎儿脐带分离、纯化和培养h UC-MSC,健康成人骨髓肝素抗凝后,采用密度梯度离心法分离、纯化和培养h BM-MSC;用倒置显微镜观察两种细胞的形态及细胞生长增殖情况,流式细胞仪分析检测第3代细胞表面标志的表达,Von?Kossa染色及油红O染色检测分化潜能。结果镜下两种细胞均为贴壁生长,形态为均一的成纤维细胞样,取相同数量的细胞传代接种后,h UC-MSC的增殖速率快于h BM-MSC,两种细胞具有均一的细胞表型,均表达CD29、CD44、CD105,不表达CD45、CD34、HLA-DR、HLA-G、CD80、CD86,两种细胞都有成骨、成脂分化潜能,但h UC-MSC的分化潜能更强。结论 h UC-MSC与h BM-MSC具有相似的生物学特性,且前者具有更强的增殖能力和分化潜能,h UC-MSC有望成为h BMMSC理想的替代来源。

Objective Human umbilical cord mesenchymal stem cells（hUC-MSC） and human bone marrow mesenchymal stem cells（hBM-MSC） were isolated, cultured and compared for their biological characteristics. Methods hUC-MSC from full-time fetus were isolated and cultured by tissue adherent method, hBM-MSC were isolated by density gradient centrifugation with heparin from a healthy adult. The morphology and proliferation of two kinds of cells were observed by a inverted microscope and the surface markers of the third-passage cells were evaluated by flow cytometry. Cell differentiation potential was evaluated with Von Kossa staining and oil red O staining. Results Both kinds of cells attached to the bottom of the wells and formed uniform fibroblast-like clusters under the microscope. The proliferation of hUC-MSC was faster than hBM-MSC in the same culture. Two kinds of cells were uniform in phenotype, expressing CD29, CD44, and CD105. They did not express CD45, CD34, HLA-DR, HLA-G, CD80 and CD86. Both cells had osteogenesis and adipogenesis differentiation potential, but the differentiation potential of hUC-MSC was higher than hBM-MSC. Conclusions hUC- MSC and hBM-MSC had similar biological characteristics, and hUC-MSC have higher proliferation and differentiation potential, hUC-MSC are expected to be an ideal alternative for hBM-MSC.