Advanced maturation by electrical stimulation: Differences in response between C2C12 and primary muscle progenitor...

Area of Science:

• Biomedical Engineering
• Regenerative Medicine
• Cell Biology

Background:

• Skeletal muscle tissue engineering faces challenges in achieving desired functional properties for regenerative medicine and meat production.
• Electrical stimulation mimics in vivo nerve stimulation, promoting muscle cell maturation in vitro.
• Translating findings from cell lines to primary cells is crucial but often debated.

Purpose of the Study:

• To investigate the effects of electrical stimulation protocols on muscle cell maturation in 2D and 3D models.
• To compare the maturation of C2C12 cell line and primary muscle progenitor cells (MPCs) in a 3D hydrogel system.
• To evaluate the translation of electrical stimulation effects from cell lines to primary cells.

Main Methods:

• Developed and applied electrical stimulation protocols to C2C12 cell monolayers (2D).
• Translated 2D protocols to a 3D model using collagen type I/Matrigel hydrogels with C2C12 and MPCs.
• Assessed muscle construct maturation via cross-striations and myosin heavy chain (MHC) isoform expression.

Main Results:

• Electrical stimulation, when optimally timed, accelerated sarcomere assembly in both 2D and 3D models.
• Muscle progenitor cell (MPC) constructs exhibited significantly higher maturation levels than C2C12 constructs.
• MPC constructs showed increased susceptibility to electrical stimulus, leading to a shift towards slower MHC isoforms.

Conclusions:

• Electrical stimulation is a viable tool for advancing skeletal muscle maturation in tissue engineering.
• Primary muscle progenitor cells (MPCs) are more responsive and achieve higher maturation levels than C2C12 cells in 3D engineered constructs.
• Optimized electrical stimulation protocols can enhance the development of functional skeletal muscle tissue, with cell source significantly impacting outcomes.