摘要
以肌组织原代培养方法对正常人成肌细胞进行体外培养；用倒置相差显微镜观察成肌细胞的成肌过程，并应用透射和扫描电镜以及免疫细胞化学技术研究成肌细胞的超微结构以及中间丝结蛋白的分布情况。结果显示，体外培养肌细胞可分为缓慢生长、分裂增殖和肌管形成期３个不同的生长、成熟阶段。成肌细胞是幼稚的早期单核肌细胞，体外分裂增殖能力强，可呈克隆生长或（和）簇状分布的不断的成熟分化过程。分裂增殖期成肌细胞胞浆有束状平行排列的肌丝以及线粒体和糖原分布，并呈逐步的分化成熟过程，可相互融合形成肌管，并可与初生肌管进一步融合发育成熟。各期成肌细胞胞浆中间丝结蛋白呈阳性反应，早期生成结蛋白量少且分布不均。上述方法对同期培养的成肌细胞与纤维母细胞可资鉴别。
Myoblast culture of 15 cases of human normal skeletal muscles were established by a modified explant technique using human fetal musle extract as a myocyte growth factor.Myogenesis was dynamically observed and morphology of myoblast was aslo preliminarily studied by phase contrast,electronmicroscopy and desmin immunocytochemistry.The results show that myocyte can be divided into three stage lag phase,divison proliferation phase and myotube formation phase when it is cultured in vitro.Myoblast was an early stage of myocyte and had a huge capacity of proliferative activity.Myoblasts clusters and clones were occasionally observed. Nascent myoblast could produce huge progeny myoblasts and they could fuse to form multinucleated myotubes Bundles of myofilaments.mitochondra,α-glycogens and free ribosomes were observed in the cytoplasm of myoblasts at the division proliferation phase in vitro.Desmin immunocytochemistry shows it significantly located in the cytoplasma of myoblasts of one,two and three weeks old cultured in vitro.Morphology of myoblast and fibroblast could be distinguished by phase contrast when they are both cultured in the late proliferation phase.
基金
福建省自然科学基金
