Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

被引:28
作者
Choi, Kwang-Hwan [1 ,2 ]
Kim, Minsu [1 ,2 ]
Yoon, Ji Won [1 ,2 ]
Jeong, Jinsol [1 ,2 ]
Ryu, Minkyung [1 ,2 ]
Jo, Cheorun [1 ,2 ,3 ]
Lee, Chang-Kyu [1 ,2 ,3 ]
机构
[1] Seoul Natl Univ, Ctr Food & Bioconvergence, Dept Agr Biotechnol, Seoul 08826, South Korea
[2] Seoul Natl Univ, Res Inst Agr & Life Sci, Seoul 08826, South Korea
[3] Seoul Natl Univ, Inst Green Bio Sci & Technol, Pyeongchang 25354, South Korea
基金
新加坡国家研究基金会; 芬兰科学院;
关键词
pig; muscle stem cells; purification; magnetic-activated cell sorting (MACS); cluster of differentiation 29 (CD29); SKELETAL; TRANSPLANTATION; PROLIFERATION; REGENERATION; MYOBLASTS; GROWTH;
D O I
10.5851/kosfa.2020.e51
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
The muscle stem cells of domestic animals are of interest to researchers in the food and biotechnology industries for the production of cultured meat. For producing cultured meat, it is crucial for muscle stem cells to be efficiently isolated and stably maintained in vitro on a large scale. In the present study, we aimed to optimize the method for the enrichment of pig muscle stem cells using a magnetic-activated cell sorting (MACS) system. Pig muscle stem cells were collected from the biceps fetnoris muscles of 14 d-old pigs of three breeds [Landracex Yorkshire xDuroc (LYD), Berkshire, and Korean native pigs] and cultured in skeletal muscle growth medium-2 (SkGM-2) supplemented with epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580). Approximately 30% of total cultured cells were nonmyogenic cells in the absence of purification in our system, as determined by immunostaining for cluster of differentiation 56 (CD56) and CD29, which are known markers of muscle stem cells. Interestingly, following MACS isolation using the CD29 antibody, the proportion of CD56(+)/CD29(+) muscle stem cells was significantly increased (91.5 +/- 2.40%), and the proportion of CD56 single-positive nonmyogenic cells was dramatically decreased. Furthermore, we verified that this method worked well for purifying muscle stem cells in the three pig breeds. Accordingly, we found that CD29 is a valuable candidate among the various marker genes for the isolation of pig muscle stem cells and developed a simple sorting method based on a single antibody to this protein.
引用
收藏
页码:852 / 859
页数:8
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