Li Xin, Cao Yang, Liu Yu, Fang Wenwen, Xiao Cheng, Cao Yang, Zhao Yumin
Institute of Animal Husbandry and Veterinary Medicine, Ji Lin Academy of Agricultural Sciences, Gongzhuling 136100, China.
Institute of Animal Biotechnology, Ji Lin Academy of Agricultural Sciences, Gongzhuling 136100, China.
Genes (Basel). 2024 Nov 21;15(12):1494. doi: 10.3390/genes15121494.
: Cultivated meat, an alternative to conventional meat, has substantial potential for alleviating environmental and ethical concerns. This method of manufacturing meat involves the isolation of skeletal muscle satellite cells (SMSCs) from donor animals, after which they proliferate in vitro and differentiate into primitive muscle fibers. The aim of this research was to evaluate how the insulin-like growth factor 1 (IGF1) gene regulates the myogenic differentiation of bovine skeletal muscle satellite cells (bSMSCs). : bSMSCs isolated from newborn calves were cultured to the third generation in vitro and differentiated into myoblasts via the serum withdrawal method. An overexpression lentivirus and siRNA targeting the IGF1 gene were constructed and transduced into bSMSCs, which were subsequently analyzed via real-time fluorescence quantitative PCR(qRT-PCR) and Western blots. The mRNA and protein levels of the myogenic differentiation markers myosin heavy chain (MyHC) and myogenin (MyoG) were determined. : The results revealed that the lentivirus overexpressing the IGF1 gene significantly increased the expression of MyHC and MyoG, whereas the expression of both the MyHC and MyoG mRNAs and proteins was strongly reduced by si-IGF1. : IGF1 positively regulates the myogenic differentiation of bSMSCs. This study provides a reference for further elucidating the molecular mechanism by which the IGF1 gene regulates the myogenic differentiation of bSMSCs via the PI3K/Akt signaling pathway and lays a foundation for establishing a regulatory network of bovine muscle growth and development.
培养肉作为传统肉类的替代品，在缓解环境和伦理问题方面具有巨大潜力。这种肉类制造方法涉及从供体动物中分离骨骼肌卫星细胞（SMSC），然后它们在体外增殖并分化为原始肌纤维。本研究的目的是评估胰岛素样生长因子1（IGF1）基因如何调节牛骨骼肌卫星细胞（bSMSC）的成肌分化。从新生小牛分离的bSMSC在体外培养至第三代，并通过血清饥饿法分化为成肌细胞。构建了靶向IGF1基因的过表达慢病毒和siRNA，并将其转导至bSMSC中，随后通过实时荧光定量PCR（qRT-PCR）和蛋白质免疫印迹法进行分析。测定了成肌分化标志物肌球蛋白重链（MyHC）和肌细胞生成素（MyoG）的mRNA和蛋白质水平。结果显示，过表达IGF1基因的慢病毒显著增加了MyHC和MyoG的表达，而si-IGF1则强烈降低了MyHC和MyoG的mRNA及蛋白质表达。IGF1正向调节bSMSC的成肌分化。本研究为进一步阐明IGF1基因通过PI3K/Akt信号通路调节bSMSC成肌分化的分子机制提供了参考，并为建立牛肌肉生长发育调控网络奠定了基础。
