Mesenchymal stem cells transduced by PLEGFP-N1 retroviral vector maintain their biological features and differentiation

作     者：HE Xu LI Yu-lin WANG Xin-rui GUO Xin NIU Yun 

作者机构：Key Laboratory of Pathobiology Ministry of Education JilinUniversity Changchun 130021 China Medical Center Hong Kong University of Science and TechnologyPeking University Shenzhen 518000 China 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2005年第118卷第20期

页      面：1728-1734页

核心收录：

学科分类：1001[医学-基础医学(可授医学、理学学位)] 100101[医学-人体解剖与组织胚胎学] 10[医学] 

基　　金：This study was supported by a grant from the National 863 Program( No.2004AA205020 )  Doctoral Foundation of Ministry ofEducation (No.20020183064) and Scientific Foundation for YoungTeachers of Jilin University (No.419070100050) 

主　　题：mesenchymal stem cells·bone marrow·enhanced green fluorescent protein·retroviral vector· gene therapy 

摘      要：Background Enhanced green fluorescent protein （EGFP） has been an important reporter gene for gene therapy. Human mesenchymal stem cells （hMSCs） are ideal target cells in cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by PLEGFP-N1 retroviral transduction. Methods hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector , and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated. Results The rate of stably transduced hMSCs-EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast-like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs. Conclusions hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy.