人脐血源性间充质干细胞的体外培养及生物学鉴定

Culture and identification of mesenchymal stem cells derived from human umbilical blood

目的建立体外培养、扩增人脐带血间充质干细胞(MSCs)的方法,初步鉴定其生物学特性。方法无菌条件下取正常足月剖宫产的脐带血,用淋巴细胞分离液分离脐血的单个核细胞,以偏酸性的MesencultTM作为培养基进行培养和纯化获得贴壁细胞层,测定MSCs的生长曲线,用流式细胞技术分析细胞的表面抗原。结果来源于脐血的单个核细胞在体外用合适的培养基培养,细胞贴壁后出现破骨样及间充质样的细胞。间充质细胞为成纤维样的细胞形态,并表达MSCs相关的抗原CD29、CD44、CD105,但不表达CD34、CD45、CD106和HLADR。这与源于骨髓的MSCs一致。结论脐带血中含有的MSCs可在体外培养、扩增,能够为实验和临床的应用提供一种新的间充质干细胞来源。

Objective To establish the method of in vitro culture and expansion of human umbilical blood-derived mesenchymal stem cells (MSCs) and to investigate their biological characteristics. Methods Human umbilical cord blood (HUCB) was collected from full term deliveries scheduled for cesarean section, the cord blood mononuclear cells were isolated by lymphocyte separation medium, purified and cultured with Mesencult TM medium and acidic environment to produce adherent layer. Growth curves of MSCs were drawn and surface antigenic features were analyzed by flow cytometry technique. Results When HUCB-derived mononuclear cells were cultured in suitable medium, the adherent cells exhibited either an osteoclast- or mesenchymal-like phenotype cells produced with the MSCs,which displayed a fibroblast-like morphology and expressed several MSCs-related antigens CD29,CD44,CD105, but not CD34,CD45,CD106, HLA-DR. The cells were identical to human bone marrow-derived MSCs. Conclusion MSCs in HUCB can be cultured and expanded in vitro, and could be regarded as an alternative source of MSCs for experimental and clinical applications.