应用小鼠早期胚胎活性因子诱导小鼠成纤维细胞再程序化为多能干细胞的实验研究  

Experimental study on inducing the mouse fibroblast reprogramming into pluripotent cells

作　　者：刘婷[1] 王立斌[1] 朱永朝[1] 金毅然[1] 李玉奎[1] 魏军[1] 

机构地区：[1]宁夏医科大学总医院宁夏人类干细胞研究所,宁夏银川750004

出　　处：《吉林医学》2013年第35期7347-7348,共2页Jilin Medical Journal

基　　金：宁夏自然科学基金资助[项目编号:NZ09133]

摘　　要：目的:研究并建立应用早期胚胎活性因子诱导成体细胞再程序化的实验体系。方法:通过体外受精方法大量收集小鼠3.5 d囊胚,从小鼠囊胚中提取早期胚胎蛋白,加入到小鼠成纤维细胞培养体系中,诱导其再程序化为多能干细胞。对得到的多能干细胞采用PCR鉴定Oct3/4基因表达;在诱导多能干细胞分化为脂肪细胞和成骨细胞后,采用油红O及茜素红染色确定其多向分化潜能。结果:诱导再程序化后获得的多能干细胞能够长期传代培养,表达干细胞特异性基因Oct3/4,采用油红O及茜素红染色确定其具有向脂肪和成骨诱导分化潜能。结论:本研究成功应用小鼠囊胚活性因子诱导小鼠成纤维细胞再程序化生成小鼠多能干细胞。Objective To search and establish the repro^amming system of the adtdt stem cell,which induced by the active factors de- rived from early mouse blastoeysts. Methods After obtaining 3.5 days mouse blastocysts by in vitro fertilization ,the embryonic protein was extracted from the blastoeysts and added to the mouse fibroblast culture system, by which reprogramming the fibroblast into pluripotent ceLls. The gene expression of Oct3/4 was tested by PCR; after induced differentiation into adipocytes and osteblasts, the eells＇ phenotype was identified by Oil Red O staining and Alizarlin Red staining. Results The induced pluripotent cells by reprog＇ramming maintained capacity of long term subeuhure; the gene of Oct3 and Oct4 were expressed in the induced pluripotent cells, which were the specific marker of stem ceils. After induced differentiation, the cells were positive stained by Oil Red O staining and Alizarlin Red staining,which were the markers of adipocyte and osteoblast. Conclusion The active factors derived from early mouse blastocysts might induce the mouse fibroblast repro- gram into the pluripotent cells. It could be provided an effective method for reprogramming of adult stem cell.

关 键 词：再程序化 多能干细胞 诱导分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]