老年大鼠骨骼肌肌源细胞的纯化及培养

The Purification and Culture of Aged Rat Skeletal Muscle Derived Cells

【目的】探讨老年大鼠骨骼肌肌源细胞纯化及培养方法。【方法】取老年雌性SD大鼠前肢肱三头肌,用胶原酶和Dispase进行消化,采用差速贴壁的方法纯化骨骼肌肌源细胞,用含20%胎牛血清的HamsF10培养基进行培养,α-骨骼肌肌动蛋白(-αsarcometric actin)免疫细胞化学染色进行细胞鉴定。【结果】成功地分离培养了老年大白鼠的骨骼肌肌源细胞。【结论】采用差速贴壁的方法可以成功纯化骨骼肌肌源细胞,为肌源细胞自体移植治疗压力性尿失禁打下了基础。

【Objective】To develope a culture and purification method of aged rat skeletal muscle derived cells.【Methods】The triceps muscle of forelimb was removed from aged SD mouse and weighed,cells were enzymatically dissociated with collagen and Dispase,and then purificated by different adhesion time.The cells were cultured in growth medium（Ham＇s F-10 nutrient mixture supplemented with 20% FBS） and identified by immunocytochemistry.【Results】Highly purified skeletal muscle derived cells were harvested successfully from aged SD and they showed α-sarcometric actin（＋） by immunocytochemistry.【Conclusion】The preplate technique can isolate and purify skeletal muscle derived cells.This study forms a basis for treatment of stress urinary incontinence using autologous muscle derived cells injection.