人胎肝MSCs的分离、鉴定及其向脂肪和成骨细胞的分化  

Isolation,identification of human fetal liver-derived mesenchymal-like stem cells and their differentiation to adipocyte and osteoblast

作　　者：王跃春[1] 张洹[1] 

机构地区：[1]暨南大学医学院血液病研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2007年第2期120-123,128,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国务院侨办重点学科建设基金(2005);暨南大学博士论文创新基金(52005031)

摘　　要：目的:分离纯化人胎肝中的间充质样干细胞(mesenchymal-like stem cells,MSCs),观察化学因子诱导其定向分化的作用,并初步鉴定其生物学特性。方法:利用二步离心法、羟乙基淀粉沉淀法及细胞差速贴壁生长特性分离纯化人胎肝MSCs;流式细胞仪检测其细胞周期和表面标志;添加常规诱导液诱导其向脂肪、成骨细胞分化并加以鉴定。结果:从人胎肝中成功分离纯化得到MSCs,P3代细胞有91.2%的细胞处于G0/G1期;表达CD29、CD44、CD105和CD166,不表达造血细胞标志CD15、CD34、CD45,不表达与GVHD相关的HLA-DR、CD80、CD86、CD40、CD40L。在经典诱导条件下,人胎肝MSCs可向脂肪及成骨细胞分化。结论:人胎肝中含有较丰富的MSCs,人胎肝MSCs具有多向分化潜能,且免疫原性弱,是组织工程和细胞治疗较为理想的种子细胞。Aim： To isolate and purify the mesenchymal-like stem cells （MSCs） from human fetal liver, and to study their biological characteristics in vitro under the induction of chemical factors. Methods： Isolate and purify the MSCs from human fetal liver based on the two-steps centrifugal way, Hespan deposition way and the different growth characteristic of attaching to the wall of cell culture flask. The cell cycle and surface markers of MSCs were identified by flow cytometry. Adding the regular inductive medium to induces the MSCs to differentiate to osteoblast and adipocyte. Osteoblast and adipocyte were identified by special staining methods. Results： The MSCs from human fetal liver expressed CD29, CD44,CD105 and CD166, but not antigens of hematopoietic CD15, CD34 ,CD45 and not antigens related to GVHD such as HLA-DR,CD80,CD86,CD40,CD40L. Exposure of these cells to osteogenic inductive agents resulted in an increase in expression of alkaline phosphatase and the appearance of hydroxyapatite nodules. Incubation with adipogenic inductive agents resulted in morphological change and positive resuits when staining with Oil Red O. Conclusion：Plentiful MSCs existed in human fetal liver. The MSCs derived from human fetal liver have multi-differentiation potency and weaker immunogenicity, so provided an ideal source of tissue engineering and cellular therapeutics.

关 键 词：人胎肝 间充质干细胞 分离 鉴定 分化 

分 类 号：R575[医药卫生—消化系统]