安淮山羊骨骼肌卫星细胞的分离培养与鉴定

发布时间：2018-12-17 02:25

【摘要】：为了分离得到安淮山羊骨骼肌卫星细胞,成功构建山羊骨骼肌卫星细胞系,为相关研究提供实验材料。以安淮山羊背最长肌肌肉组织为材料,采取胶原酶和胰蛋白酶结合的二步消化法,结合差速贴壁法纯化原代骨骼肌卫星细胞。对骨骼肌卫星细胞中特异性基因Pax7用免疫荧光染色法及PCR扩增加以鉴定。通过分离、纯化,得到纯度较高的山羊骨骼肌卫星细胞,其形态为圆形,折光性强。培养一段时间后密度增大,细胞呈梭型。所得骨骼肌卫星细胞生长状态良好,具有较为一致的生长方式和形态特点。经免疫荧光染色法以及PCR鉴定发现细胞系中有Pax7基因的表达。通过上述材料和方法,可获得较纯的山羊骨骼肌卫星细胞,并稳定传代。
[Abstract]:In order to isolate and obtain Anhuai goat skeletal muscle satellite cells, a goat skeletal muscle satellite cell line was successfully constructed. Using the longissimus dorsi muscle tissue of Anhuai goat as the material, the primary skeletal muscle satellite cells were purified by the two-step digestion method combined with collagenase and trypsin, and the differential adhesion method was used to purify the primary skeletal muscle satellite cells. The specific gene Pax7 in skeletal muscle satellite cells was identified by immunofluorescence staining and PCR amplification. High purity goat skeletal muscle satellite cells were obtained by isolation and purification. After a period of culture, the density of the cells increased and the cells were fusiform. The satellite cells of skeletal muscle are in good growth state and have consistent growth pattern and morphological characteristics. The expression of Pax7 gene was identified by immunofluorescence staining and PCR. Through the above materials and methods, more pure goat skeletal muscle satellite cells can be obtained and subcultured stably.
【作者单位】：安徽农业大学动物科技学院;安徽地方畜禽遗传资源保护与生物育种省级实验室;安徽省羊繁育工程技术研究中心;
【基金】：安徽省自然科学基金项目(1708085MC61) 安徽省现代农业产业技术体系(2016-2020)共同资助
【分类号】：S827

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