重庆医学2013，Vol.42Issue(1)：8-12,5.DOI:10.3969/j.issn.1671-8348.2013.01.003
SOX9基因修饰骨髓间充质干细胞并诱导其向髓核样细胞分化的实验研究
Experimental research of SOX9 gene in modifying marrow mesenchymal stem cells and inducing their differentiation toward nucleus pulposus-like cells
许运 1陈亮 2史勇 1顾勇 2邹俊 2唐天驷2
作者信息
- 1. 苏州市相城人民医院骨科,江苏苏州,215131
- 2. 苏州大学附属第一医院骨科,江苏苏州,215006
- 折叠
摘要
Abstract
Objective To investigate lentivirus vector containing SOX9 gene infecting rabbit marrow mesenchymal stem cells(MSC)and inducing their differentiation toward nucleus pulposus-like cells .Methods Lentivirus vectors containing SOX9 gene were constructed and were used to infect MSC .MSC were divided into none transduction group ,blank transduction group and SOX9 transduction group .Flow cytometry was employed to detect the infection rate and apoptotic rate of MSC ,MTT was adopted to measure the MSC proliferation .RT-PCR ,Western blot and immunofluorescent staining were applied to detect the mRNA or protein expression of SOX9,Collagen type E and Aggrecan in MSC .Results Lentivirus vector containing SOX9 gene was successfully constructed and infected MSC with infection rate of 100% .There was no significant impact of lentivirus vector infection on proliferation and apoptosis of MSC .After 2 weeks infection ,mRNA and protein expression levels of SOX9 and Collagen type E in MSC in SOX9 transduction group were markedly higher than those in none transduction group and blank transduction group (P< 0 .01) ,while mRNA or protein expression of Aggrecan only existed in MSC in SOX9 transduction group.Collagen type Ⅱ- and Aggrecan- positive fluorescent staining were found in MSC in SOX9 transduction group,while those in none transduction group and blank transduction group showed negative .Conclusion SOX9 lentiviral vector infected MSC can be induced to differentiate into nu -cleus pulposus-like cells .关键词
间质干细胞/慢病毒感染/细胞分化/胶原Ⅱ型/基因,SOX9Key words
mesenchymal stem cells /lentivirus infections /cell differentiation /collagen type Ⅱ /gene,SOX9引用本文复制引用
许运,陈亮,史勇,顾勇,邹俊,唐天驷..SOX9基因修饰骨髓间充质干细胞并诱导其向髓核样细胞分化的实验研究[J].重庆医学,2013,42(1):8-12,5.
