人脐血间充质干细胞体外分离培养与扩增实验研究

目的探讨脐血来源的人间充质干细胞（HMSCs）在体外分离培养与扩增的可行性。方法在无菌条件下收集正常足月胎儿的脐带血，经复合枸橼酸钠抗凝，用相对密度为1．077g／L的Ficoll淋巴细胞分离液分离脐血的单个核细胞，以30％胎牛血清进行培养和扩增，用流式细胞仪检测MSCs的表面标志。结果来源于人脐血的单个核细胞接种于特定培养基后，可产生贴壁细胞，主要表现为破骨样细胞和间充质样细胞，经传几代后，可得纯化扩增的人脐血MSCs。流式细胞仪检测结果显示，人脐血MSCs不表达GD14、CD19，强表达CD105、CD44。结论来源于人脐血的MSCs在体外可以分离培养、扩增，为MSCs的进一步研究奠定基础。

Isolation and identification of mesenchymal stem cells from human umbilical cord blood in vitro

Objective To investigate the feasibility and optimal condition of isolation and amplification of mesenchymal stem cells from human umbilical cord blood （HUCB） in vitro. Methods Human umbilical cord blood was collected, anticoagulanted by sodium citrate compound ,isolated by density gradient centrifugation,,these cells were proliferated clonally i,dected surface signs of MSCS by flow cytometry. The cord mononuclear cells were isolated with lymphocyte separation medium （density 1.077 g/ml）,purified and expanded with specific medium ,the surface antigen expression of MSCs was detected with flow eytometry. Results The Human umebilical cord blood derived mononuclear cells,when seeded in specific medium ,gave rise to adherent cells ,which exhibited either an osteoclast or mesenchymal-like phenotype, after three passage, these cells were able to be purified and expanded. MSCs did not express antigens CD14 and CD19,but express strongly CD105 and CD44. Conclusion MSCs in HUCB can be cultured and amplified in vitro and could lay the foundation for further study.