非整合型质粒重编程脐带血CD34^+细胞形成诱导性多潜能干细胞的研究  

Efficient Reprogramming of Human Cord Blood CD34+ Cells for Formation of Induced Pluripotent Stem Cells with Non-integrating Plasmid System

作　　者：张权娥 刘淑平[1] 刘延风[1] 张鸿雁[1] 袁卫平[1] 陈桂彬[1] 李彦欣[1] 许静[1] 

机构地区：[1]中国医学科学院,北京协和医学院血液病医院(血液学研究所)实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2013年第3期728-734,共7页Journal of Experimental Hematology

基　　金：国家医药重大专项(编号2011ZX09102-010-04);科技部973项目(编号2012CB966601;2010CB945204;2009CB521803);国家自然科学基金(编号81130074;81070390);天津市科委自然基金项目(编号10JCZDJC19500;11JCZDJC27900)

摘　　要：本研究探索和优化非整合质粒的方法,将人脐带血来源的CD34+(CB CD34+)细胞进行重编程,建立无病毒的iPS技术体系。利用细胞核转染仪将质粒pEB-C5和pEB-Tg转入短暂培养后的CB CD34+细胞中,使其进行重编程形成iPSC,14 d后观察到2×106CB CD34+细胞中约有200个类似ES细胞特征的克隆出现,并对产生的iPSC进行体内外多能性鉴定及细胞核型检测。结果表明,重编程后的CB CD34+细胞的多能性基因表达与ESC类似,细胞核型正常,外源基因无插入,且具有体内分化成三胚层的全能性。结论:利用非整合型质粒的方法重编程CB CD34+细胞形成iPSC,该方法无外源基因插入、重复性好、操作简单、且效率较高,为建立相对安全的iPSC提供了一个行之有效的途径,为深入探索iPSC应用于临床药物的筛选、组织工程和再生医学研究等提供了很好的研究材料。This study was to establish the episomal vector reprogramming method to reprogram iPSC from human cord blood （CB） CD34 cells. The non-integrating plasmids of pEB-C5 and pEB-Tg were transfected into short-term cultured CB CD34 cells by using the nucleofector, so as to demonstrate efficient reprogramming of CB CD34 cells. Within 14 days of one-time transfection by two plasmids together, up to 200 iPSC-like colonies per 2 million transfected CB CD34 cells were generated. The results showed that the pluripotency of iPSC-derived CB CD34 cells was similar to that of hESC and the karyotypes of iPSC were normal. In addition, no vector integration was found in iPSC of 9th and 10th passages. Furthermore, hiPSC formed teratoma with three embryonic germ layers. It is concluded that the integration-free method to generate human iPSC from CB CD34 cells is reliable and can provide new ways for both research and future clinical applications.

关 键 词：多潜能干细胞 脐带血CD34+细胞 非整合型质粒 

分 类 号：R331.1[医药卫生—人体生理学]