成年动物骨骼肌细胞原代培养及转染外源基因的初步研究

PRIMARY CULTURE OF ADULT SKELETAL MUSCLE CELLS AND EXPRESSION OF TRANSFECTED FOREIGN GENES

目的 观察损伤对成年动物骨骼肌卫星细胞增殖的影响 ,以获取基因治疗自体移植的载体。 方法 以阳离子脂质体介导法将 p RSV- L ac Z或 p Rch- TH质粒转入培养肌细胞。 结果 骨骼肌损伤后 ,分离出的肌卫星细胞及组织块游离成肌细胞数均显著增高 ;外源基因能在细胞内表达。 结论 损伤可诱导成年骨骼肌卫星细胞增殖并促进体外培养肌细胞成活。

Objective\ To stimulate the proliferation of the muscle cells from adult animals with a slight injury for primary culture, in order to get the genetically modified cells for autografting. Methods The injury of the gastrocnemius muscle of the animal was repeatedly made by a short period of blockage of the popliteal artery with a rubber band and an electro\|stimulation was simultaneously given to the muscle. This injured muscle was then taken out from the leg and cultured for 72 hrs. These muscle cells were then transfected by a plasmid containing a report gene, LacZ gene, or objective gene, TH gene, by using the method of lipofection in vitro. Results The primary muscle cells were survived properly and grew luxuriantly. An appearance of proliferation of muscle cells was also seen in the culture. Quite a few of these muscle cells showed a good expression of LacZ or TH gene with X\|gal or TH staining after transfecting. Conclusion The results indicate that the skeletal muscle cell from an adult animal, including human being, can be cultured in vitro after a slight injury and this provides a useful way to practice gene therapy.