A modified preplate technique for efficient isolation and proliferation of mice muscle-derived stem cells

被引:6
|
作者
Xu, Zhuqiu [1 ]
Yu, Lu [1 ]
Lu, Haibin [1 ]
Feng, Weifeng [1 ]
Chen, Lulu [1 ]
Zhou, Jing [1 ]
Yang, Xiaonan [1 ]
Qi, Zuoliang [1 ]
机构
[1] Chinese Acad Med Sci, Plast Surg Hosp, Peking Union Med Coll, Beijing 100041, Peoples R China
基金
中国国家自然科学基金;
关键词
Regenerative medicine; Cell and tissue culture; Myogenic stem cells; In vitro culture; SKELETAL-MUSCLE; PROGENITOR CELLS; SATELLITE CELLS; IN-VITRO; IDENTIFICATION; POPULATION; DIFFERENTIATION; TRANSPLANTATION; HETEROGENEITY; REGENERATION;
D O I
10.1007/s10616-018-0262-0
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We modified an existing protocol to develop a more efficient method to acquire and culture muscle-derived stem cells (MDSCs) and compared the characteristics of cells obtained from the two methods. This method is based on currently used multistep enzymatic digestion and preplate technique. During the replating process, we replaced the traditional medium with isolation medium to promote fibroblast-like cell adherence at initial replating step, which shortened the purifying duration by up to 4days. Moreover, we modified the culture container to provide a stable microenvironment that promotes MDSC adherence. We compared the cell morphology, growth curve and the expression of specific markers (Sca-1, CD34, PAX7 and Desmin) between the two cell groups separately obtained from the two methods. Afterwards, we compared the neural differentiation capacity of MDSCs with other muscle-derived cell lineages. The protocol developed here is a fast and effective method to harvest and purify MDSCs from mice limb skeletal muscle.
引用
收藏
页码:1671 / 1683
页数:13
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