Maintenance of human embryonic stem cells on gelatin  

作　　者：LI Yang LIN ChangSheng WANG Li LIU Ying MU XiaoNing MAYue LI LingSong 

机构地区：[1]Peking University Stem Cell Research Center and China National Center for International Research, Beijing 100191, China [2]Department of Reproduction, Beijing Obstetrics and Genecology Hospital, Beijing 100026, China [3]The Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China

出　　处：《Chinese Science Bulletin》2009年第22期4214-4220,共7页

基　　金：Supported by the National Key Basic Research and Development of China (Grant Nos. 2006CB943603 and 2006CB503905);International Collaboration, the Ministry of Science and Technology of China (Grant No. 20070192)

摘　　要：Matrigel is routinely used as a coating material in the feeder-free culture system of human embryonic stem cells (hESCs). However, matrigel is costive and inconvenient to use. In this study, the possibility of using gelatin as an alternative coating material was investigated. The results showed that, after trypsinization, hESCs were maintained undifferentiated on gelatin. These hESCs expressed pluripotent markers, formed teratoma and maintained a normal karyotype. As measured at passage 10, the hESCs expressed a high level of Oct4 on both gelatin and Matrigel. hESCs growing on gelatin formed AP-positive colonies in similar size and number to those growing on Matrigel (P > 0.05). Moreover, hESCs growing on gelatin contained a comparable percentage of SSEA-4-positive cells to those growing on Matrigel (95.1% vs.94.3%, P > 0.05). H-1 hESCs were maintained undifferentiated on gelatin for 20 passages and remained the stable normal karyotype. This gelatin-based culture protocol may allow us to propagate hESCs in large scale, with less cost.Matrigel is routinely used as a coating material in the feeder-free culture system of human embryonic stem cells （hESCs）. However, matrigel is costive and inconvenient to use. In this study, the possibility of using gelatin as an alternative coating material was investigated. The results showed that, after trypsinization, hESCs were maintained undifferentiated on gelatin. These hESCs expressed pluripotent markers, formed teratoma and maintained a normal karyotype. As measured at passage 10, the hESCs expressed a high level of Oct4 on both gelatin and Matrigeh hESCs growing on gelatin formed AP-positive colonies in similar size and number to those growing on Matrigel （P〉 0.05）. Moreover, hESCs growing on gelatin contained a comparable percentage of SSEA-4-positive cells to those growing on Matrigel （95.1% vs.94.3%, P〉 0.05）. H-1 hESCs were maintained undifferentiated on gelatin for 20 passages and remained the stable normal karyotype. This gelatin-based culture protocol may allow us to propagate hESCs in large scale, with less cost.

关 键 词：人类胚胎干细胞 明胶 维护 涂层材料 培养体系 胰蛋白酶 阳性克隆 阳性细胞 

分 类 号：Q813[生物学—生物工程] TQ431.3[化学工程]