论文部分内容阅读
目的 研究脐带血 (HUCB)中含有的间充质干细胞 (MSCs)体外分离、纯化及培养条件 ,探究其作为种子细胞应用于实验和临床的可行性。方法 无菌条件下取正常足月剖腹产的脐带血 ,经肝素抗凝 ,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 ,用流式细胞仪检测MSCs表面抗原。结果 来源于脐血的单个核细胞经体外培养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 (CD2 9、CD4 4、CD1 6 6 ) ,但不表达造血细胞抗原 (CD34 、CD4 5) ,这与源于骨髓的MSCs一致。结论 脐带血来源的MSCs能在体外培养、扩增 ,而用于满足实验和临床的需要
Objective To study the isolation, purification and culture conditions of mesenchymal stem cells (MSCs) contained in umbilical cord blood (HUCB) in vitro and explore the feasibility of using them as seed cells in experimental and clinical research. Methods Umbilical cord blood of normal full-term cesarean section was obtained under aseptic conditions. After heparin anticoagulation, mononuclear cells of umbilical cord blood were separated by lymphocyte separation liquid, cultured and purified with MesencultTM as acidic medium to obtain adherent cell layer, MSCs surface antigens were detected by flow cytometry. Results The mononuclear cells derived from umbilical cord blood had osteoclast-like and mesenchymal-like cells after in vitro culture. The mesenchymal cells were fibroblast-like and expressed MSCs-related antigens (CD29, CD4 4, CD166), but not hematopoietic antigen (CD34, CD4 5), which is consistent with bone marrow-derived MSCs. Conclusion MSCs derived from umbilical cord blood can be cultured and expanded in vitro, and can be used to meet the experimental and clinical needs