转基因人骨髓间质干细胞在支架中增殖及矿化的研究

Proliferation and mineralization of human mesenchymal stem cells on scaffold

目的:观察携带骨形态发生蛋白2基因(BMP-2)的人骨髓间质干细胞(hMSCs)在生物衍生骨支架环境中的增殖及矿化。方法:利用BMP-2腺病毒载体转染hMSCs后种植生物衍生骨支架中,扫描电镜观察细胞生长增殖,能量谱仪测定钙质分泌,RTPCR检测培养液上清中BMP-2表达。结果:转染后,hBMSC呈ALP、Ⅰ和Ⅱ型胶原阳性表达;细胞在支架中伸展良好,生长旺盛,并产生钙盐沉积;RTPCR显示培养液上清中存在BMP-2基因表达。结论:转染BMP2基因的hMSCs在生物衍生骨支架中能够立体生长、增殖,并继续表达目的基因,为基因修饰的组织工程骨修复骨缺损奠定了基础。

Objective:To study proliferation and mineralization of human mesenchymal stem cells (hMSCs)carrying bone morphogenetic protein-2 gene (BMP-2)on allogenetic bone scaffold.Method:After infected with adenoviral vector carrying BMP-2 gene,hMSCs were implanted into allogenetic bone scaffold.Cellular proliferation was observed by electron microscope scanning;calcium content secretion was measured by energy spectrometer;BMP-2 expression in culture supernatant was analyzed by reverse transcription-polymerase chain reaction(RT-PCR).Result: After infection,hMSCs expressed positively ALP,collagen type Ⅰ and type Ⅱ.The cells grew well and produced calcium deposit on the scaffold.BMP-2 gene expression was detected in the culture supernatant by RT-PCR.Conclusion:hMSCs transfected by BMP-2 gene can grow and proliferate three-dimensionally,and continue to express exogenous gene,which lay a solid foundation for gene modified tissue engineering bone to repair bone defects.