Enhanced growth and myogenic differentiation of spheroid-derived C2C12 cells

Biosci Biotechnol Biochem. 2021 Apr 24;85(5):1227-1234. doi: 10.1093/bbb/zbab018.

Abstract

Among many factors of controlling stem cell differentiation, the key transcription factor upregulation via physical force is a good strategy on the lineage-specific differentiation of stem cells. The study aimed to compare growth and myogenic potentials between the parental cells (PCs) and the 1-day-old C2C12 spheroid-derived cells (SDCs) in two-dimensional (2D) and three-dimensional (3D) culture conditions through examination of the cell proliferation and the expression of myogenic genes. The data showed that 1-day-old spheroids had more intense expression of MyoD gene with respect to the PCs. The proliferation of the SDCs is significantly higher than the PCs in a time-dependent manner. The SDCs had also significantly higher myogenic potential than the PCs in 2D and 3D culture conditions. The results suggest that MyoD gene upregulation through cell-cell contacts is the good approach for preparation of seed cells in muscle tissue engineering.

Keywords: 2D and 3D culture; C2C12; cell expansion; myogenesis; spheroids.

MeSH terms

  • Actinin / genetics
  • Actinin / metabolism
  • Animals
  • Cell Culture Techniques*
  • Cell Differentiation
  • Cell Line
  • Cell Proliferation / drug effects
  • Collagen / chemistry
  • Collagen / pharmacology
  • Gene Expression Regulation
  • Mice
  • Muscle Cells / cytology
  • Muscle Cells / drug effects
  • Muscle Cells / metabolism*
  • Muscle Development / drug effects
  • Muscle Development / genetics*
  • MyoD Protein / antagonists & inhibitors
  • MyoD Protein / genetics*
  • MyoD Protein / metabolism
  • Myoblasts / cytology
  • Myoblasts / drug effects
  • Myoblasts / metabolism*
  • Myogenin / genetics
  • Myogenin / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Spheroids, Cellular / cytology
  • Spheroids, Cellular / drug effects
  • Spheroids, Cellular / metabolism*
  • Tissue Engineering / methods

Substances

  • MyoD Protein
  • MyoD1 myogenic differentiation protein
  • Myog protein, mouse
  • Myogenin
  • RNA, Small Interfering
  • Actinin
  • Collagen