基础医学与临床2013，Vol.33Issue(7)：892-897,6.
人胚胎干细胞无血清无饲养层培养体系的建立
Establishment of a serum-free and feeder-free culture system for human embryonic stem cells
摘要
Abstract
Objective To establish a serum-free and feeder-free culture system for human embryonic stem cells (HESc).Methods The HESX-01 cells were cultured in Knockout medium (the first group),self-made serumfree medium (the second group) and improved serum-free medium with bFGF and heparin (the third group),respectively.The morphology,clone counting and clone size were observed after twenty-fifth generation.The surface labeling immunocytochemical analysis,Fluorescence-activated cell sorter analysis (FACS) and embryoid bodies formation were utilized to determine the character of HESc.Results After twenty-fifth passages the clones of the second group were gradually differentiated,and the HESX-01 cells in the first and third group showed typical human embryonic stem cell morphology,they all expressed Nanog,Oct-4,Tra-1-60 and SSEA-4,but did not express SSEA-1,the same results were observed from FACS analysis.The HESc formed embryoid body.Besides,the significant differences were oberserved in the second group as compared with the first and third group in clone counting and size per field.Conclusions The self-made serum-free and feeder-free culture system for HESX-01 cells has been established,and the efficacy is not significantly different to commercially availabe Knockout medium.关键词
人胚胎干细胞/自制无血清/无饲养层/bFGF/肝素Key words
human embryonic stem cells/ self-made serum-free/ feeder-free/ bFGF/ heparin分类
医药卫生引用本文复制引用
喻松霞,郭玲玲,贺文凤,贺玲,李洁,李剑..人胚胎干细胞无血清无饲养层培养体系的建立[J].基础医学与临床,2013,33(7):892-897,6.基金项目
国家自然科学基金(30760261) （30760261）
国家大学生创新性实验计划(07-17) （07-17）
江两省青年科学家培养对象计划项目(2007DQ0014) （2007DQ0014）
