Isolation and characterization of human fetal myoblasts.
Isolation and characterization of human fetal myoblasts.
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DOI：
10.1007/978-1-61779-343-1_1
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Gussoni, Emanuela
中科院分区：
文献类型：
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作者：
Lapan, Ariya D;Gussoni, Emanuela
分离的人胎儿骨骼肌含有肌源性细胞，以及非肌源性细胞，如脂肪细胞、成纤维细胞和淋巴细胞。因此，重要的是确定一种有效和可靠的分离方法，以获得更纯的成肌细胞群体。为此，荧光激活的细胞分选结合稳健的肌原细胞表面标记物可用于富集解离肌肉中的成肌细胞。在这一章中，我们描述了一种方法，显着丰富的成肌细胞使用黑色素瘤细胞粘附分子（MCAM），我们已经确定是一个很好的标记人胎儿成肌细胞。然后，这种分离方法产生的成肌细胞可以在体外扩增，并且仍然保留显着的生肌活性，如体外融合试验所示。从分离的肌肉中分离高度肌源性群体的能力有利于骨骼肌发育和肌肉疾病的体外研究。此外，这些细胞的稳健扩增将为人类肌肉疾病的细胞疗法的开发带来新的见解。
Dissociated human fetal skeletal muscle contains myogenic cells, as well as non-myogenic cells such as adipocytes, fibroblasts, and lymphocytes. It is therefore important to determine an efficient and reliable isolation method to obtain a purer population of myoblasts. Toward this end, fluorescence-activated cell sorting in conjunction with robust myogenic cell surface markers can be utilized to enrich for myoblasts in dissociated muscle. In this chapter, we describe a method to significantly enrich for myoblasts using melanoma cell adhesion molecule (MCAM), which we have determined to be an excellent marker of human fetal myoblasts. The myoblasts resulting from this isolation method can then be expanded in vitro and still retain significant myogenic activity as shown by an in vitro fusion assay. The ability to isolate a highly myogenic population from dissociated muscle facilitates the in vitro study of skeletal muscle development and muscle diseases. Furthermore, robust expansion of these cells will lead to new insights in the development of cell-based therapies for human muscle disorders.