Simplifying the protocol for low-pollution-risk, efficient mouse myoblast isolation and differentiation

  • Adv Biotechnol (Singap). 2025 Mar 11;3(1):8. doi: 10.1007/s44307-025-00060-0.
Yi Luo  #  1 ,  Jia-Dong Zhang  #  1 ,  Xue-Gang Zhao  2 ,  Wei-Cai Chen  1 ,  Wan-Xin Chen  1 ,  Ya-Rui Hou  1 ,  Ya-Han Ren  1 ,  Zhen-Dong Xiao  1 ,  Qi Zhang  1  3 ,  Li-Ting Diao  4 ,  Shu-Juan Xie  5  6
Affiliations
  • 1. Biotherapy Center, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China.
  • 2. Department of Surgical Intensive Care, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China.
  • 3. Vaccine Research Institute of Sun Yat-Sen University, Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China.
  • 4. Biotherapy Center, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China. [email protected].
  • 5. Biotherapy Center, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China. [email protected].
  • 6. Vaccine Research Institute of Sun Yat-Sen University, Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510630, China. [email protected].
  • # Contributed equally.
PMID: 40067554 DOI: 10.1007/s44307-025-00060-0
Abstract

Myoblasts are the primary effector cells that play crucial roles in myogenesis and muscle regeneration following injury. However, isolating purified primary myoblasts from murine skeletal muscle poses challenges for junior researchers. Here, we present a simplified, low-risk, and optimized protocol for the extraction and enrichment of these myogenic progenitor cells. Additionally, we demonstrate that, compared to F10 (Ham's F-10)-based medium, DMEM (Dulbecco's Modified Eagle's Medium)-based differentiation medium provides a more conducive environment for myoblasts differentiation. This enhancement improves the efficiency of myofiber formation and the expression of myogenic markers.

Keywords
Differentiation; Isolation; Primary myoblasts.
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