PcDNA3-hBMP2转染骨髓基质细胞及其稳定表达

目的：通过观察转染hBMP-2基因的骨髓基质细胞的生长特性的变化和检测目的基因在受体细胞中表达，探讨骨髓基质细胞用作hBMP-2基因的受体细胞的可能性。方法：在脂质体介导下将hBMP-2基因导入体外培养的骨髓基质细胞中，用G418筛选获得阳性细胞，分别应用原位杂交技术和酶联免疫吸附方法检测目的基因在受体细胞内的存在与表达。结果：在mRNA水平可检测到目的基因在阳性细胞中进行了转录，在培养基中，用ELISA的方法能检测到目的基因在骨髓基质细胞中进行了表达，并分泌到培养基中。从细胞的生长曲线上，可知细胞的倍增时间并没有因为目的基因的导入而改变，其生长时间也与正常的细胞相近。结论：骨髓基质细胞可以用作hBMP-2基因的受体细胞，表达并分泌hBMP-2蛋白质，也可作为骨组织工程学中的种子细胞。

Transfection of bone marrow stromal cells with PcDNA3-hBMP2 and expression detection of hBMP-2 gene

Objective: To explore the feasibility of bone marrow stromal cells (BMSCs) being used as accepted cells of hBMP-2 gene by studying the instantaneous mRNA and its protein expression of hBMP-2 in rabbit's BMSCs, and to make further study on the effects of transfection on biological behavior of these cells. Methods: With the help of the lipofectamine , the cultured BMSCs were transfected with PcDNA3-hBMP2 and the positive cells were screened out by G418. The evidence of successful transfection in these cells could be obtained by in situ hybridization and ELISA. Results: After transfection with PcDNA3-hBMP2, hBMP-2 gene's transcribed product- mRNA can be found in the positive cells ,and hBMP-2 proteins can be examined in the culture . The biological behavior of the positive cells unchanged after the target gene was transfected into the cells. Conclusions: The BMSCs could be used as the accepted cells of hBMP-2 gene for expressing and secreting the hBMP-2 into the culture, which can be served as seed cells in bone tissue engineering .