摘要
目的利用 Ad5/CMV－ TGF－β 1转染体外培养兔骨髓间充质干细胞，测定 TGF－β 1的表达产物为关节软骨缺损的基因治疗提供实验基础。方法采用体外培养法对兔骨髓间充质干细胞进行原代培养，应用已构建好的 Ad5/CMV－ TGF－β 1真核载体对其进行转染，并用免疫组织化学方法对转染细胞进行表达产物测定。结果采用腺病毒转染技术能够成功的实现骨髓间充质干细胞的外源性基因修饰，没有发生明细的细胞毒性反应； Ad5/CMV－ TGF－β 1对骨髓间充质干细胞的转染率为 95％左右；转染 8周后仍能观察到 TGF－β 1的表达。结论采用真核表达载体 Ad5/CMV－ TGF－β 1可以成功转染体外培养的骨髓间充质干细胞并可获得 TGF－β 1的高表达。
Objective Bone marrow mesenchymal stem cells(BMSCs) are transfectde with Ad5/CMV－ TGF－β 1, then the expression of TGF－β 1, are evaluted to provide the experimented foundation of gene therapy in treating joint articular defect. Methods BMSCs were obtained from bone marrow of 4 adult Newzealand rabbit and culture. Primary cultures were transfected with Ad5/CMV－ TGF－β 1, then the expression of TGF－β 1 protein were determined with immunohistochemical method. Result Adeno－ virus transduction technology could sucessfully complete the exogene modification of BMSCs, no obvious cell toxic reaction. The transduction ration of Ad5/CMV－ TGF－β 1 to BMSCs were 95％ and the exogenous gene expression could sustain for over 8 weels. Conclusion Ad5/CMV－ TGF－β 1 can be transfected to BMSCs and the expression of TGF－β 1 can be transfected to BMSCs and the expression of TGF－β 1 is high.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
2001年第20期42-43,共2页
Journal of Clinical Rehabilitative Tissue Engineering Research
