基础医学与临床2012，Vol.32Issue(1)：71-76,6.
用组织贴壁法从整根脐带分离培养间充质干细胞及其生物学特性的检测
Isolation and characterization of mesenchymal stem cells derived from whole human umbilical cord by applying a direct explant technique
摘要
Abstract
Objective To establish a technique for isolating mesenchymal stem cells from umbilical cord and their biological characteristics. Methods Mesenchymal stem cells (MSCs) were isolated from umbilical cord(UC) by applying a explant technique and explanding cultured in vitro. Growth curves were drawn, and The phenotypes and cell cycle were evaluated by flow cytometry. UC-MSCs were induced to differentiate into adipocytes, osteocytes and chondrocytes in special differentiation condition. Mutidifferentiation related genes and stem cell-related transcription factors, Nanog, Oct-4 Sox-2 were detected by TR-PCR. Results UC-MSCs were isolated using a explant technique , and characteristic of plastic adherence and fibroblast-like morphology, the adherent cells displayed an abundant presence of CD73, CD90, CD105 and absence of CD34, CD45, HLA-DR. Cell cycle showed that there werepercentages of stem cells as Go/G1 81. 56 % and S + G2 + M 18. 44% respectively. When cultured in differentiation media, they differentiated into adipocytes, osteocytes and chondrocytes. RT-PCR reactions confirmed that their mutidifferentiation related genes were positive. Moreover, stem cell-related transcription factors, Nanog, Oct-4 Sox-2 were positively expressed in UC-MSCs. Conclusions The explant technique is a feasible method to obtain MSCs from whole human umbilical cord, and UC can be considered as a novel and convenient source of adult MSCs displaying primitive pluripotent sten cells and high expansion potential.关键词
脐带/间充质干细胞/分化Key words
umbilical cord/mesenchymal stem cell/differentiation分类
医药卫生引用本文复制引用
张颢,张斌,程梅,陶艳玲,扈江伟,徐曼,陈虎..用组织贴壁法从整根脐带分离培养间充质干细胞及其生物学特性的检测[J].基础医学与临床,2012,32(1):71-76,6.基金项目
国家高技术研究发展计划项目(863项目)(2011AA020114) （863项目）
军队临床高新技术重大项目(2010gxjs100) （2010gxjs100）
黑龙江省卫生厅科研课题(2011-518) （2011-518）
首都临床特色应用研究(Z111107058811107) （Z111107058811107）
哈尔滨市科技创新人才研究专项基金(2009RFQQS027) （2009RFQQS027）
