SDF-1 cDNA转染人骨髓间充质干细胞的实验研究

Experimental Study on Human Bone Marrow Mesenchymal Stem Cells Transfected by SDF-1 cDNA

本研究旨在探讨基质细胞衍生因子-1(SDF-1)cDNA瞬时转染人骨髓间充质干细胞(hBMSCs)前后hBMSCs中SDF-1mRNA表达的情况。在分离、培养、鉴定人骨髓间充质干细胞(hBMSCs)和构建SDF-1-pIRES2-EGFP真核表达载体的基础上,通过脂质体介导法将SDF-1真核表达质粒转染至hBMSCs,观测绿色荧光蛋白的表达并用RT-PCR方法检测瞬时转染效率。结果发现:SDF-1-pIRES2-EGFP瞬时转染后hBMSCs的SDF-1mRNA表达增高约20%左右。结论:阳离子脂质体可以将SDF-1cDNA真核表达质粒瞬时转染入hBMSCs,但转染效率低下,不能获得足够数量的稳定转染细胞供后续实验,因此需对转染方法作进一步的探讨。

This study was aimed to investigate the expression of SDF-1 mRNA in SDF-1 cDNA-modified human bone marrow mesenchymal stem cells （hBMSCs） before and after transfection. The hBMSCs were isolated, cultured and identified, the SDF-1-pIRES2-EGFP eukaryotic expressing vector was constructed, and then the hBMSCs were transfected with the vector encapsulated by lipofectamineTM 2000. The transfection efficiency was measured by observing the expression of green fluorescence protein and detecting the mRNA by RT-PCR. The results indicated that the expression of SDF-1 mRNA increased by about 20% after hBMSCs were transfected instantaneously by SDF-1-pIRES2- EGFP. It is concluded that SDF-1 cDNA eukaryotic expression vector can be instantly transfected into hBMSCs by lipofectamineTM 2000, but the efficiency was too low to obtain enough steady transferred hBMSCs. Other procedures should be trialed to improve the transfection efficiency.