外周血红系祖细胞来源诱导多能干细胞的建立  

Generation of induced pluripotent stem cells from erythroid progenitor cells in circulating peripheral blood

作　　者：梁淑贞[1] 林茂[1] 陈继冰[1] 牛立志[1] 

机构地区：[1]暨南大学医学院附属复大肿瘤医院/广州复大肿瘤医院中心实验室,广州510655

出　　处：《临床检验杂志》2016年第8期597-600,共4页Chinese Journal of Clinical Laboratory Science

基　　金：广州市海珠区科技计划项目(2014-cg-05)

摘　　要：目的建立和鉴定外周血中红系祖细胞来源的诱导多能干细胞(induced pluripotent stem cells,i PSCs)。方法从健康人外周血标本中分选红系祖细胞,将表达Oct4、Sox2、Lin28、L-Myc和Klf4转录因子的ori P/EBNAl附着体电转染红系祖细胞使其重编程获得i PSCs,并通过核型鉴定、碱性磷酸酶染色、RT-PCR反应、畸胎瘤形成实验及类胚体形成实验检测其干细胞多能性特性。结果获得的i PSCs核型正常,碱性磷酸酶染色呈阳性,表达干细胞多能性基因Sox2、Oct4、Nanog和Klf4和Lin28,体内畸胎瘤形成实验可分化为内、中、外三胚层细胞,体外可形成类胚体。结论成功建立外周血红系祖细胞来源具有多向分化潜能的i PSCs。Objective To generate and identify induced pluripotent stem cells （iPSCs） from erythroid progenitor cells in circulating peripheral blood（PB）. Methods Erythroid progenitor cells were isolated from PB of healthy individuals and were infected by episomal vectors with oriP/EBNA-1 （ Epstein-Barr nuclear antigen-1 ） backbone for delivering the reprogramming genes Oct,l, Sox2, Lin28, L- Myc and Klf4 to generate iPSCs. The multipotency of the obtained iPSCs was verified by karyotyping analysis, alkaline phosphatase staining, RT-PCR, teratoma test, and the formation of embryoid body. Results The obtained iPSCs carried normal karyotype and expressed pluripotent genes Sox2, Oct4, Nanog, Klf4 and Lin28. Alkaline phosphatase staining of the iPSCs showed positive result. In the teratoma test, the cells from the three germ layer, ectoderm, mesoderm and endoderm, were all found. Embryoid bodies were formed in vitro in suspended culture of the iPSCs. Condusion The iPSCs from erythroid progenitor cells in circulating peripheral blood with muhipotency were successfully generated.

关 键 词：红系祖细胞 转录因子 重编程 诱导多能干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]