南方医科大学学报2015，Vol.35Issue(8)：1090-1096,7.
骨髓间充质干细胞体外培养及肝向分化方案的优化
Optimization of the protocols for in vitro culture and induction of hepatic differentiation of rat mesenchymal stem cells
摘要
Abstract
Objective To optimize the protocols for isolation, in vitro culture, identification and induction of hepatic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were separated and purified by differential adherent culture for 1.5 h with the first medium change at 12 h. The surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into adipogenic, osteogenic, and chondrogenesis lineages. A 3-step protocol including sequential addition of growth factors, cytokines and hormones was used to induce the BMSCs to differentiate into hepatocyte-like cells. Results The cells isolated using this protocol were positive for CD29, CD44, and CD90 and negative for CD29 and CD45. The adipogenic, osteogenic, and chondrogenic differentiation of the BMSCs were verified by Oil red, Alizarin red, and toluidine blue staining. The BMSCs induced with the 3-step protocol differentiated into hepatic-like cells that expressed hepatocyte-specific proteins (ALB and AFP) and genes. Conclusion The optimized protocol allows simple and efficient isolation of highly purified populations of BMSCs, which can be induced into hepatic lineages in specific microenvironment.关键词
骨髓间充质干细胞/体外培养/肝向分化Key words
bone marrow mesenchymal stem cell/culture in vitro/hepatic differentiation引用本文复制引用
向俊西,郑幸龙,祝旭龙,杨丽斐,高睿,李建辉,刘学民,吕毅..骨髓间充质干细胞体外培养及肝向分化方案的优化[J].南方医科大学学报,2015,35(8):1090-1096,7.基金项目
教育部高等学校博士学科点专项科研基金(20110201130009) （20110201130009）
陕西省科学技术研究发展计划国际合作项目(2012KW-39-01) （2012KW-39-01）
