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目的探讨hPDGF-A/hBD2腺病毒双基因转染大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)后,对BMSCs本身生物学特性的影响。方法体外分离培养大鼠BMSCs,应用GFP标记的腺病毒表达载体系统Adv-hPDGF-A-IRES-hBD2,以脂质体法转染293细胞,再以病毒上清液感染BMSCs,对转染目的基因后的BMSCs进行形态学观察,绘制生长曲线,测定其细胞周期以及向成脂、成骨、成肌诱导分化和鉴定。结果hPDGF-A/hBD2双基因成功转入BMSCs后,未发现其对细胞活性有明显作用,基因修饰后的BMSCs仍具有成体干细胞所具备的增殖能力,以及向成脂、成骨、成肌等方向分化的潜能。结论BMSCs是重组腺病毒转染的良好靶细胞,hPDGF-A/hBD2基因修饰的BMSCs仍可作为满意的细胞治疗的种子细胞。
Objective To investigate the effect of hPDGF-A / hBD2 adenovirus double transfection on the biological characteristics of BMSCs after transfection of bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were isolated and cultured in vitro. Ad-hPDGF-A-IRES-hBD2 was transfected into 293 cells by lipofection method using GFP-labeled adenovirus vector. BMSCs were infected with virus supernatant. After the gene, the BMSCs were observed morphologically, the growth curve was drawn, and the cell cycle and the differentiation and identification to adipogenic, osteogenic and myogenic differentiation were determined. Results The hPDGF-A / hBD2 double gene was successfully transfected into BMSCs without any significant effect on the cell viability. Gene-modified BMSCs still have the ability of proliferation of adult stem cells, as well as adipogenic, osteogenic and myogenic The potential for differentiation. Conclusion BMSCs are good target cells for transfection of recombinant adenovirus. BMSCs modified by hPDGF-A / hBD2 gene can still be used as seed cells for satisfactory cell therapy.