Sox9基因真核表达载体的构建及其诱导大鼠骨髓基质细胞定向分化为骨骺干细胞Construction of Sox9 eukaryotic expression vector and its inductive effect on marrow stroma cells differentiating into precartilaginous stem cells
胡伟华;陈安民;郭风劲;李锋;黄晖;张伟凯;
摘要(Abstract):
[目的]从永生化骨骺干细胞中克隆Sox9基因并构建真核表达载体,并探讨Sox9诱导骨髓基质细胞向骨骺干细胞分化的可能性。[方法]以RT-PCR方法获得Sox9全长,插入pGEM-TEasy克隆载体中,测序正确后与pEG-FP-IRES2表达载体酶切后连接,复合质粒以脂质体法转染骨髓基质细胞,观察转染效率,Sox9和FGFR-3的表达。流式细胞术鉴定细胞表型,MTT法检测细胞增殖活性。[结果]成功的完成了Sox9的扩增和表达载体的构建,重组载体转染骨髓基质细胞后能检测到Sox9、FGFR-3的表达,增殖活性与骨骺干细胞无异。[结论]成功构建了Sox9真核表达载体,其能诱导骨髓基质细胞分化为骨骺干细胞并具有其特性。
关键词(KeyWords): 骨骺干细胞;Sox9;骨髓基质细胞;转染
基金项目(Foundation): 国家自然科学基金资助项目(编号:30650006);; 教育部博士点基金资助项目[No.教技发中心函(2005)216]
作者(Author): 胡伟华;陈安民;郭风劲;李锋;黄晖;张伟凯;
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参考文献(References):
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