骨骼肌卫星细胞体外培养及分化为成骨样细胞的实验研究

Study on the Differentiation of Skeletal Muscle Satellite Cells Transfected by Ad-BMP2 Gene into Osteoblast-like Cells

目的:评估骨骼肌卫星细胞(skeletalmusclesatellitecells,SMSCs)体外分化为成骨样细胞的能力,探讨骨骼肌卫星细胞作为骨组织工程种子细胞的可能性。方法:取新生Wistar大鼠骨骼肌,采用酶消化法分离出SMSCs,体外原代及传代培养,经腺病毒介导的人骨形成蛋白2(Ad-BMP2)基因转染后,行成骨细胞标志物活性的检测及细胞化学染色,并进行细胞体外矿化能力的测定。结果:转染后细胞的碱性磷酸酶(ALP)活性增强(P<0.01)且组织化学染色呈阳性,骨钙素及I型胶原的免疫细胞化学染色呈阳性,21d后见钙结节形成。结论:体外培养的骨骼肌卫星细胞可以向成骨方向转化,有望成为骨组织工程的种子细胞。

Objective： To assess the ability of inducing skeletal muscle satellite cells（SMSCs） into osteoblast-like cells, and to approach the possibility of treating skeletal muscle satellite cells as a new seed cell for bone tissue engineering. Method： The skeletal muscles of newborn Wistar rats were acquired to separate skeletal muscles satellite cells with en- zyme digestion. Cells are cultured and subcultured in vitro. After being transfected by Ad-BMP2 gene, cells were examined by osteoblastic marker＇s activity test, cytochemical staining and mineralized capability in vitro. Results： After the cells were transfected, there was a significant increase in ALP activity（P〈0.01）, the cells were faint positive with immunocytochemical stain of osteocalcin and type 1 collagen. Calcium nodules were seen after cells were cultured for 21 days. Conclusion： SMSCs have the capacibility to be differentiated into osteoblast and to be regarded as the seed cells for bone tissure engineering.