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背景:目前对于大鼠、恒河猴等骨骼肌卫星细胞的研究较多,但有关小鼠骨骼肌卫星细胞的研究报道较少。目的:观察体外培养的小鼠骨骼肌卫星细胞在成肌过程中的光电镜结构及免疫组织化学特性。设计:单一样本研究。单位:天津中医学院组织胚胎学教研室。材料:实验在北京大学基础医学院人体解剖学与组织胚胎学系唐军民导师研究室完成。新生5d昆明种小鼠5只,雌雄不限(购自北京大学医学部实验动物中心)。干预:分离新生小鼠骨骼肌卫星细胞,进行体外培养。在倒置显微镜下观察肌卫星细胞的成肌过程,并应用透射电镜及免疫细胞化学技术观察小鼠成肌细胞的超微结构及结构蛋白的分布。主要观察指标:①小鼠骨骼肌卫星细胞的分离纯化与原代培养的形态学观察。②免疫组织化学观察。结果:卫星细胞生长、分化良好,最终由单个核圆形细胞逐渐变化为多个核的长圆柱的肌管状细胞。免疫细胞化学技术显示,细胞呈α-肌动蛋白阳性。结论:采用两步消化法分离,得到的骨骼肌卫星细胞质量较好;且α-肌动蛋白对鉴定小鼠骨骼肌卫星细胞具有特异性。
BACKGROUND: At present, there are many researches on skeletal muscle satellite cells in rats and rhesus monkeys, but there are few reports about the satellite cells in mouse skeletal muscle. OBJECTIVE: To observe the light microscopic structure and immunohistochemical characteristics of mouse skeletal muscle satellite cells cultured in vitro during myogenic development. Design: Single Sample Study. Unit: Department of Embryology, Tianjin College of Traditional Chinese Medicine. MATERIALS: Experiments were performed at the Institute of Human Anatomy and Histology and Embryology, Peking University Institute of Materia Medica Tang Junmin. Newborn 5d Kunming mice 5, male or female (purchased from Peking University Health Science Laboratory Animal Center). Intervention: Isolation of neonatal mouse skeletal muscle satellite cells for in vitro culture. The muscle satellite cells were observed under an inverted microscope. The ultrastructure and the distribution of structural proteins of the myoblasts were observed by transmission electron microscopy and immunocytochemistry. MAIN OUTCOME MEASURES: ① Isolation and purification of mouse skeletal muscle satellite cells and morphological observation of primary culture. ② immunohistochemical observation. Results: The satellite cells grew well and differentiated well. Eventually, the long cylindrical myotubes of multiple nuclear nuclei gradually changed from single nuclei to round nuclei. Immunocytochemistry showed that the cells were α-actin positive. Conclusion: Skeletal muscle satellite cells obtained by two-step digestion method are of good quality. Α-actin is specific to the identification of mouse skeletal muscle satellite cells.