Isolation and culture of multifidus satellite cells by single muscle fiber method

doi:10.3969/j.issn.2095-4344.0941

Abstract

Abstract:

BACKGROUND: Atrophy and damage of multifidus muscles mainly result in various low back pains and dysfunctions. Muscle satellite cells, which belong to muscle stem cells, play an important role for muscle regeneration after injury. However, little is reported on the isolation and culture of multifidus muscle up to now.
OBJECTIVE: To isolate and identify multifidus satellite cells of rats using single muscle fiber method.
METHODS: Multifidus muscles were isolated in vivo and multifidus satellite cells were obtained by single muscle fiber method. Then the cells were purified by repeated differential attachment. Morphological changes of the cells were observed and the proliferation curve was made thereafter. Moreover, myogenic differentiation of the isolated cells cultured in differentiation medium was analyzed and marker proteins of multifidus satellite cells (Pax7, Myod, Desmin) were characterized by immunofluorescence.
RESULTS AND CONCLUSION: A large number of fusiform or spindle cells were seen around the muscle fibers after 72 hours of culture. Round cells with high refraction were visible after being purified by repeated differential attachment, and the cells adhered after 24-48 hours of re-inoculation exhibited an “S” shaped growth curve. Large myotubes formed in the differentiation medium. Immunofluorescence findings showed that Pax7 and Myod were positively expressed in muscle satellite cell nuclei, while Desmin was positively expressed in the cytoplasm. These findings reveal that multifidus satellite cells can be successfully separated by single muscle fiber method, which can be used for regeneration of multifidus muscles in the future.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Satellite Cells, Skeletal Muscle, Muscle Fibers, Skeletal, Cell Separation, Tissue Engineering

CLC Number:

R394.2

Liu Tong, Yu Jia-ni, Chen Yu-pei, Chen Huan, Zou De-hui, Yan Jun, Lu Zong-xiao, Liu Yue, Zhang Li, Huo Ze-jun. Isolation and culture of multifidus satellite cells by single muscle fiber method[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(25): 4065-4070.

Figures/Tables 1

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