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目的建立一种快速高效的小鼠成体骨骼肌卫星细胞的提取、纯化的实验方法。方法通过布比卡因预处理激活肌卫星细胞,48小时后,以混合酶一步消化法及改良纯化法从成年骨骼肌中分离高纯度卫星细胞,并与普通差速贴壁法、反复差速贴壁法及Percoll非连续密度梯度离心法等纯化方法相比较。结果通过改良提取、纯化法所得到的骨骼肌卫星细胞纯度可达到(82.53±3.16)%,明显高于普通差速贴壁法、反复差速贴壁法及Percoll非连续密度梯度离心法(P<0.05),并具有很好的增殖及分化能力。结论通过采用改良成体骨骼肌卫星细胞提取及纯化法,能快速高效地获取高纯度肌卫星细胞。
Objective To establish a rapid and efficient experimental method for the extraction and purification of adult mouse skeletal muscle satellite cells. Methods Muscle satellite cells were activated by bupivacaine preconditioning. After 48 hours, high purity satellite cells were isolated from adult skeletal muscle by one-step enzyme digestion and modified purification, and compared with ordinary differential adherence method, repeated differential Adherent method and Percoll non-continuous density gradient centrifugation and other purification methods compared. Results The purity of skeletal muscle satellite cells obtained by improved extraction and purification method was (82.53 ± 3.16)%, which was significantly higher than that of ordinary differential adhesion method, repeated differential adhesion method and Percoll discontinuous density gradient centrifugation method (P <0.05), and has good proliferation and differentiation ability. Conclusion High purity myoelectric satellite cells can be quickly and efficiently obtained by using the method of extracting and purifying satellite cells from adult skeletal muscle.