激活素A促进人胚胎干细胞定向分化为视网膜色素上皮细胞  

Activin A promotes human embryonic stem cells to directly differentiate into retinal pigment epithelium cells

作　　者：李奇[1,2] 王磊[2] 祝贺[2] 吴骏[2] 郝捷[2] 王柳[2] 周琪[1,2] 

机构地区：[1]吉林大学动物科学学院动物胚胎工程吉林省重点实验室,长春130062 [2]中国科学院动物研究所干细胞与生殖生物学国家重点实验室,北京100101

出　　处：《科学通报》2016年第16期1816-1821,共6页Chinese Science Bulletin

基　　金：中国科学院战略性先导科技专项(XDAO01020101);中国科学院动物研究所干细胞与生殖生物学国家重点实验室SKLRB青年杰俊科研创新主任基金资助

摘　　要：视网膜色素上皮细胞(retinal pigment epithelium,RPE)的功能紊乱、退化和丢失导致的感光细胞的损伤是造成老年性黄斑变性(age-related macular degeneration,AMD)和视网膜色素变性(retinitis pigmentosa,RP)的主要原因.目前临床常规治疗只能延缓患者视力减退的进程,无法根治该类疾病.近期的临床研究表明,视网膜下腔移植RPE能够改善患者视力,这为RPE细胞用于临床治疗提供了很好的依据.由于原代RPE不能再生且资源有限,所以可以通过人胚胎干细胞(human embryonic stem cells,h ESCs)分化获得RPE.为解决h ESCs自发分化获得RPE的效率很低、分化时程较长的问题,在常规分化RPE的前7天加入激活素A(Activin A),结果发现Activin A能够促进h ESCs向RPE的分化.鉴定结果表明,分化获得的细胞表达RPE相关蛋白:OTX2,MITF和BEST1,以及RPE特异标记物:MITF和RPE65,而且具有正常的形态特征和吞噬功能.上述结果表明分化得到的细胞具有RPE的基本特性.该分化方法能够为后期的研究和临床治疗提供大量的细胞.Retinal pigment epithelium（RPE） is a kind of monolayer cells situated between neural retina and choroid,and plays an important role in maintaining the functions of retina and photoreceptor. Its functions mainly include absorbing stray light,providing nutrients for neural retina and phagocytizing outer section of photoreceptor. Photoreceptor damage caused by dysfunctions,degeneration and loss of RPE is the main reason that leads to age-related macular degeneration（AMD） and retinitis pigmentosa（RP）. After subretinal transplantation of differentiated RPE into the rat model with retinal degeneration disease,visual functions and light sensors have been improved. At present,the conventional clinical treatments can only delay the process of patient＇s visual deterioration,and cannot cure the disease. Recent clinical studies showed that cell therapy based on subretinal transplantation RPE could promote patient＇s vision,which provide a good basis for the RPE replacement therapy. Because primary RPE is not renewable and its resource is limited,so human embryonic stem cells（h ESCs） were used to be differentiated into RPE. However spontaneous differentiation of h ESCs into RPE is low efficient,and the process is long,so improving differentiation efficiency of RPE is of large urgency. Transforming growth factor-b（TGF-b） superfamily adjusts RPE formation during the period of embryonic development,adding TGF-b into differentiation system can promote the RPE differentiation. Activin A is a member of the TGF-b superfamily. In this study,Activin A was added in the first 7 days of conventional differentiation of RPE,and according the results of number counting of pigment cluster Activin A could promote the differentiation of h ESCs into RPE. Next,identification of the h ESCs derived RPE was performed. Under the optical microscope,the cells showed a primary RPE-like hexagonal morphology. Furthermore,the h ESCs derived RPE expressed PRE specific markers： MITF,RPE65,OTX2 and BEST1,which were confirmed using th

关 键 词：激活素A 人胚胎干细胞 视网膜色素上皮细胞 定向分化 

分 类 号：R774[医药卫生—眼科]