人类胚胎来源成肌细胞的培养和鉴定  

Culture and identification of human embryo-derived myoblasts

作　　者：刘桂英[1] 杨立业[2,3] 李文玉[2] 郑佳坤[3] 陈强[2] 

机构地区：[1]潮州市中心医院口腔科,广东省潮州市521021 [2]潮州市中心医院检验医学中心,广东省潮州市521021 [3]潮州市中心医院,神经外科,广东省潮州市521021

出　　处：《中国组织工程研究》2013年第32期5806-5812,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省自然科学基金项目(020001,04007929);广东省卫生厅立项课题(A2003886);潮州市科委基金资助项目;中国博士后科学基金资助(2004036491)~~

摘　　要：背景:人类胚胎骨骼肌含有成肌细胞,其在体外条件下的培养及在体外能否融合形成肌管,以及表达的相应的标志物,目前尚不明确。目的:验证源于人类胚胎骨骼肌的成肌细胞在体外条件下的培养条件,能否在体外融合形成肌管,能否表达神经细胞的标志物。方法:采用组织块培养法对人类胚胎肌肉来源的成肌细胞进行原代培养,以免疫细胞化学染色检测培养的细胞肌肉细胞标志物desmin、myogenin、平滑肌肌动蛋白、myosin和神经细胞标志物β-tubulinⅢ、nestin、neurofilament200(NF200)、胶质纤维酸性蛋白的表达。结果与结论:从人类胚胎肌肉组织中成功培养出成肌细胞,表达成肌细胞的标志物desmin和myogenin,同时也表达神经元特异性烯醇化酶、nestin和NF200,细胞能够在体外融合形成含有多个细胞核的肌管,融合的肌管可以表达NF200、β-tubulinⅢ和胶质纤维酸性蛋白等神经细胞的标志物。结果证实,人类胚胎肌肉来源的成肌细胞能够同时表达神经细胞和肌肉细胞的标志物,培养的成肌细胞和肌管细胞表达神经元特异性烯醇化酶、β-tubulinⅢ、nestin、NF200和胶质纤维酸性蛋白。说明这几种神经细胞标志物不能用于肌肉来源的细胞向神经细胞跨分化的鉴定研究。BACKGROUND： There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE： To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS： Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cells were identified with immunocytochemistry for neural markers, such as β-tubulin Ⅲ, nestin, neuron specific enolase, neurofilament 200, and glial fibrillary acidic protein, and muscle markers （desmin, myogenin, smooth muscle actin and myosin）. RESULTS AND CONCLUSION： A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressed βⅢ-tubulin, neurofilament 200 and glial fibrillary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase, β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cell identification of trans-differentiation study from muscle origin to nervous system.

关 键 词：干细胞 胚胎干细胞 成肌细胞 胚胎 肌管 分化 细胞培养 胶质纤维酸性蛋白 神经元特异性烯醇化酶 省级基金 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]