구.농수식품
- 조회1,377
Jinsol Jeong1, Kwang-Hwan Choi1,2, Seung-Hun Kim1, Dong-Kyung Lee1,2, Jong-Nam Oh1, Mingyun Lee1, Gyung Cheol Choe1 and Chang-Kyu Lee1,3*
1Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture and Life Science, Seoul National University, Seoul 08826, South Korea.
2Present address: Research and Development Center, Space F corporation, Hwasung-si, Gyeonggi-do 18471, South Korea.
3Institute of Green Bio Science and Technology, Seoul National University, Pyeong Chang, Kangwon-do 25354, South Korea.
* Correspondence
Abstract
Background
Myogenic transdifferentiation can be accomplished through ectopic MYOD1 expression, which is facilitated by various signaling pathways associated with myogenesis. In this study, we attempted to transdifferentiate pig embryonic fibroblasts (PEFs) myogenically into skeletal muscle through overexpression of the pig MYOD1 gene and modulation of the FGF, TGF-β, WNT, and cAMP signaling pathways.
Results
The MYOD1 overexpression vector was constructed based on comparative sequence analysis, demonstrating that pig MYOD1 has evolutionarily conserved domains across various species. Although forced MYOD1 expression through these vectors triggered the expression of endogenous muscle markers, transdifferentiated muscle cells from fibroblasts were not observed. Therefore, various signaling molecules, including FGF2, SB431542, CHIR99021, and forskolin, along with MYOD1 overexpression were applied to enhance the myogenic reprogramming. The modified conditions led to the derivation of myotubes and activation of muscle markers in PEFs, as determined by qPCR and immunostaining. Notably, a sarcomere-like structure was observed, indicating that terminally differentiated skeletal muscle could be obtained from transdifferentiated cells.
Conclusions
In summary, we established a protocol for reprogramming MYOD1-overexpressing PEFs into the mature skeletal muscle using signaling molecules. Our myogenic reprogramming can be used as a cell source for muscle disease models in regenerative medicine and the production of cultured meat in cellular agriculture.
논문정보
- Research article
- 2021년 05월 (BRIC 등록일 2021-06-08)
- 국내 연구진
- 생명과학 > 동물자원학
- 구・농수식품
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