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该文探讨了5、10、15、21、42日龄大鼠骨骼肌卫星细胞(muscle-derived satellite cells,MDSCs)生物学特性。利用二次酶消化法提取MDSCs原代细胞。利用不同的诱导培养液使MDSCs定向分化为神经细胞、成骨细胞和肌细胞,并通过特异性染色和免疫组化法对其进行鉴定。结果显示,5、10、15、21日鼠龄大鼠骨骼肌中均可高效分离出原代MDSCs且细胞活力旺盛,42日大鼠MDSCs原代分离困难。以上结果一定程度上拓展了MDSCs的取材范围。第5、10、15、21日大鼠来源MDSCs经成神经细胞诱导后呈多角形、有明显的树突结构且表达神经特异性烯醇化酶(neuron-specific enolase,NSE),证明MDSCs可以分化为神经细胞;经成骨诱导后细胞形成明显的骨节结,茜素红及骨钙蛋白(osteocalcin)染色均呈阳性;经成肌诱导后细胞发生融合形成成熟肌管细胞且表达快肌肌球蛋白。出生后5、10、15、21日大鼠来源MDSCs取材范围广,易于体外分离培养,增殖能力强且具有一定的多能性,适合作为再生医学的种子细胞。
This article explored the biological characteristics of 5, 10, 15, 21, 42-day-old rat skeletal muscle-derived satellite cells (MDSCs). Primary culture of MDSCs cells by secondary enzymatic digestion. MDSCs were differentiated into neurons, osteoblasts and myocytes by using different induction culture medium, and identified by specific staining and immunohistochemistry. The results showed that the primary MDSCs could be efficiently isolated from rat skeletal muscle of 5, 10, 15, 21 days and the viability of the cells was good. The primary isolation of MDSCs on the 42nd was difficult. The above results to a certain extent, expanded the scope of the draw of MDSCs. On the 5th, 10th, 15th and 21st days, MDSCs derived from rat were polygonal with induced dendritic structure and expressed neuron-specific enolase (NSE), demonstrating that MDSCs could differentiate Which were neurons. After osteogenic induction, the cells formed obvious osteodules, which were all positive by alizarin red and osteocalcin staining. After myogenic induction, the cells fused to form mature myotubes and express the myofibers protein. After birth, the MDSCs derived from rats at 5, 10, 15 and 21 days were obtained from a wide range of sources and were easily isolated and cultured in vitro with strong proliferative ability and pluripotency and were suitable as seed cells for regenerative medicine.