中国实验动物学报2011，Vol.19Issue(4)：271-276,6.DOI:10.3969/j.issn.1005-4847.2011.04.001
人脐带间充质干细胞作为维持人胚胎干细胞生长饲养层细胞的研究
Human umbilical cord mesenchymal stem cells support prolonged undifferentiated growth of human embryonic stem cells
摘要
Abstract
Objective To find a new source of human feeder cells which can support the undifferentiated growth of human embryonic stem cells. In this study,human umbilical cord mesenchymal stem cells( HUCMSCs) were tested as feeder cells for supporting the growth of hESCs to prevent the possibility of contamination from animal feeder cells. Methods Mesenchymal stem cells were isolated from human umbilical cord by collagenase IV digestion. Their surface markers were detected by flow cytometry. The cells were induced to differentiate into osteoblast and adipocytes. HESc line H1 was grown on inactivated human umbilical cord mesenchymal stem cells, and were passaged every five days . After many passages, the morphology, expression of specific markers and differentiation capacity of hESCs were examined. Results HUCMSCs were spindle-shaped adherent cells. They had strong proliferation ability, and showed strongly positive expression of CD44,CD29,CD73,CD105,CD90,CD86,CD147,CD117 and negative expression of CD14, CD38, CD133, CD34.CD45 and HLA-DR. After induction, oil red 0 staining was positive. After passaged on MSCs feeder cells for 20 passages, hESCs maintained their morphological characteristics. Immunofluorescence staining showed positive expression of OCT4, Nanog, SSEA4, TRA-1-60, and TRA-1-81 and negative SSEA1. Alkaline phosphatase staining showed positive results. Moreover, the hECSs cultured on HUCMSCs formed embryoid body in vitro confirmed their pluripotency. Conclusions HUCMSCs can be used as the feeder cells for hESCs culture and maintain their undifferentiated state.关键词
人胚胎干细胞/小鼠胚胎成纤维细胞/人脐带间充质干细胞/饲养层细胞/临床应用Key words
Human embryonic stem cells/ Mouse embryonic fibroblast cells/Human umbilical cord mesenchymal stem cells/ Feeder cells/ Clinical application分类
生物科学引用本文复制引用
刘峰,徐永胜,俞海燕,王薇,张纯..人脐带间充质干细胞作为维持人胚胎干细胞生长饲养层细胞的研究[J].中国实验动物学报,2011,19(4):271-276,6.基金项目
国家自然科学基金(编号:30772376) （编号:30772376）
北医三院临床重点项目基金(编号:BYSY200903) （编号:BYSY200903）
教育部高等学校博士学科点专项科研基金(编号:20100001120100). （编号:20100001120100）
