摘要
背景:移植间充质干细胞预防和治疗椎间盘退变是一种可行的方法,将SOX-9和GDF-5共同转染骨髓间充质干细胞,使其向髓核细胞转化,以期获得更大的髓核诱导和促增殖效应。目的:探讨SOX-9和GDF-5基因共同诱导兔骨髓间充质干细胞向类髓核细胞分化的效果。方法:提取、分离、纯化4周龄新西兰大白兔骨髓间充质干细胞,取第3代骨髓间充质干细胞分为5组体外诱导其向类髓核细胞分化,分别为未转染组、空载体转染组、SOX-9转染组、GDF-5转染组、共转染组。转染后第14天采用RT-PCR检测SOX-9,GDF-5和Ⅱ型胶原的m RNA表达,免疫组化染色法检测髓核细胞标记物KRT19表达。结果与结论:共转染组SOX-9 m RNA表达高于转染SOX-9组,差异有显著性意义(P<0.05);共转染组GDF-5m RNA表达高于转染GDF-5组,差异有显著性意义(P<0.05)。共转染组Ⅱ型胶原表达高于转染SOX-9组、转染GDF-5组,差异有显著性意义(P<0.05)。SOX-9转染组及GDF-5转染组KRT19呈阳性表达,共转染组呈强阳性表达,可见被转染的骨髓间充质干细胞向类髓核细胞分化,且双基因转染诱导骨髓间充质干细胞向类髓核细胞分化的能力和分泌细胞外基质的能力明显高于单基因转染。
BACKGROUND：Transplantation of mesenchymal stem cels to prevent and treat degeneration of the intervertebral disc is a feasible method. Mesenchymal stem cels co-transfected by SRY-related high mobility group-box gene 9 （SOX-9） and growth differentiation factor-5 （GDF-5） can differentiate into nucleus pulposus cels, in order to obtain greater effect of induction and proliferation of nucleus pulposus cels. OBJECTIVE：To investigate the effect of SOX-9 and GDF-5 co-transfection on the differentiation of rabbit bone marrow mesenchymal stem cels into nucleus pulposus cels. METHODS： We separated and cultured bone marrow mesenchymal stem cels from the bone marrow of rabbit aged 4 months. Passage 3 cels were divided into five groups andin vitro induced to differentiate into nucleus pulposus cels： non-transfected group, empty vector transfection group, SOX-9 transfection group, GDF-5 transfection group, SOX-9 and GDF-5 co-transfection group. At 14 days after transfection, RT-PCR was employed to assay SOX-9, GDF-5 and colagen type II mRNA expressions in bone marrow mesenchymal stem cels. The marker of nucleus pulposus cels-KRT19 expression was also detected by immunohistochemical staining. RESULTS AND CONCLUSION：In the co-transfection group, the mRNA expressions of SOX-9, GDF-5, and colagen type II were significantly higher than those in the SOX-9 transfection group, GDF-5 transfection group, and both these two groups, respectively （P ＆lt; 0.05）. Cels were positive for KRT19 in the SOX-9 and GDF-5 groups, and strongly positive for KRT19 in the co-transfection group. These findings indicate that double gene-transfected bone marrow mesenchymal stem cels are better than single gene-transfected cels with regard to differentiation into nucleus pulposus cels and secretion of extracelular matrix.
出处
《中国组织工程研究》
CAS
北大核心
2015年第19期2953-2958,共6页
Chinese Journal of Tissue Engineering Research
基金
内蒙古自然科学基金(2009MS1117)~~
