不同鼠龄大鼠骨骼肌卫星细胞生物学特性的探讨  

Study on the Biological Characteristics of Rat Skeletal Muscle-Derived Satellite Cells from Rats in Different Age

作　　者：任宇 俞兰 赵兴胜 

机构地区：[1]内蒙古自治区人民医院,呼和浩特010017

出　　处：《中国细胞生物学学报》2017年第5期566-574,共9页Chinese Journal of Cell Biology

基　　金：内蒙古自治区自然科学基金项目(批准号:2016BS0314);内蒙古自治区人民医院院内基金(批准号:2016012)资助的课题~~

摘　　要：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 study was aimed to study on the biological characteristics of rat skeletal muscle-derived satellite cells（MDSCs） in rats aged 5, 10, 15, 21, 42 days. MDSCs were separated and cultured by double enzyme digestion and differential adherent culture. Neurogenic, osteogenic and myogenic induction media were used to induce directed differentiation. Differentiated nerve cells, osteoblasts and myotubes cells were identified by morphology and immunohistochemical staining, respectively. Primary MDSCs of the 5th, 10th, 15th and 21 st day were highly efficiently separate from homologous skeletal muscles with a wide range of source. It was difficult to separate the MDSCs from 42 nd day rat. After neurogenesis, four groups MDSCs formed neuron-specific enolase positive polygonal-shaped dendritic cells. It is the first time to identified that MDSCs can be differentiated into neural cells. After osteogenesis, MDSCs in rats aged 5, 10, 15, 21, 42 days formed Alizarin red-and osteocalcin-positive bone nodules. After myogenesis, myotubes were formed and expressed fast muscle myosin-positive. MDSCs of the 5th, 10th, 15th and 21 st day are easy to isolate culture and amplify in vitro with pluripotency, which increases the range of source material available for tissue engineering and are suitable as seed cells for regenerative medicine.

关 键 词：骨骼肌卫星细胞 鉴定 成神经细胞诱导 成骨诱导 成肌诱导 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]