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Updated: Aug 7, 2026
Preparation and Culture of Myogenic Precursor Cells/Primary Myoblasts from Skeletal Muscle of Adult and Aged Humans
Published on: February 16, 2017
NANOG restores the impaired myogenic differentiation potential of skeletal myoblasts after multiple population
Aref Shahini1, Debanik Choudhury1, Mohammadnabi Asmani2
1Bioengineering Laboratory, Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Amherst, NY 14260-4200, USA.
Abstract:
Adult skeletal muscle regeneration relies on the activity of satellite cells residing in the skeletal muscle niche. However, systemic and intrinsic factors decrease the myogenic differentiation potential of satellite cells thereby impairing muscle regeneration. Here we present data showing that late passage C2C12 myoblasts exhibited significantly impaired myogenic differentiation potential that was accompanied by impaired expression of myogenic regulatory factors (Myf5, MyoD, Myogenin, and MRF4) and members of myocyte enhancer factor 2 family. Notably, ectopic expression of NANOG preserved the morphology and restored the myogenic differentiation capacity of late passage myoblasts, possibly by restoring the expression level of these myogenic factors. Muscle regeneration was effective in 2D cultures and in 3D skeletal microtissues mimicking the skeletal muscle niche. The presence of NANOG was required for at least 15days to reverse the impaired differentiation potential of myoblasts. However, it was critical to remove NANOG during the process of maturation, as it inhibited myotube formation. Finally, myoblasts that were primed by NANOG maintained their differentiation capacity for 20days after NANOG withdrawal, suggesting potential epigenetic changes. In conclusion, these results shed light on the potential of NANOG to restore the myogenic differentiation potential of myoblasts, which is impaired after multiple rounds of cellular division, and to reverse the loss of muscle regeneration.
Insights
NANOG protein can restore muscle regeneration potential in aged cells. This discovery offers hope for reversing age-related decline in muscle repair and function.
Area of Science:
- Muscle biology
- Cellular regeneration
- Stem cell research
Background:
- Adult skeletal muscle regeneration depends on satellite cells.
- Aging and other factors reduce satellite cell function, impairing muscle repair.
- Late-passage myoblasts show decreased myogenic differentiation and impaired gene expression.
Purpose of the Study:
- To investigate the role of NANOG in restoring myogenic differentiation in aged myoblasts.
- To explore NANOG's potential to reverse impaired muscle regeneration.
Main Methods:
- Using C2C12 myoblast cell cultures at late passages.
- Ectopic expression of NANOG in myoblasts.
- Assessing myogenic differentiation capacity and gene expression (Myf5, MyoD, Myogenin, MRF4, MEF2).
- Utilizing 2D cultures and 3D skeletal microtissues.
Main Results:
- Late-passage myoblasts exhibited reduced myogenic potential and impaired expression of key myogenic factors.
- Ectopic NANOG expression restored myoblast morphology and differentiation capacity.
- NANOG presence for 15 days reversed impaired differentiation, but removal was necessary for myotube formation.
- NANOG-primed myoblasts retained differentiation capacity for 20 days post-withdrawal, suggesting epigenetic modifications.
Conclusions:
- NANOG can restore myogenic differentiation potential in aged myoblasts.
- NANOG may reverse the loss of muscle regeneration capacity associated with cellular aging.
- Findings suggest NANOG's potential therapeutic role in muscle repair and regeneration.
