The Effects of Muscle Cell Aging on Myogenesis

Abstract

The process of myogenesis gradually deteriorates as the skeletal muscle ages, contributing to muscle mass loss. The aim of this study is to investigate the effect of senescence/aging on skeletal myogenesis, in vitro. A model of multiple cell divisions of C2C12 myoblasts was used to replicate cell senescence. Control and aged myoblasts were investigated during myogenesis, i.e., at days 0, 2, and 6of differentiation. SA-β-gal activity and comet assay were used as markers of aging and DNA damage. Flow cytometry was performed to characterize potential differences in cell cycle between control and aged cells. Alterations in the mRNA and/or protein expression of myogenic regulatory factors (MRFs), IGF-1 isoforms, apoptotic, atrophy, inflammatory, metabolic and aging-related factors were evaluated. Compared with the control cells, aged myoblasts exhibited G0/G1 cell cycle arrest, DNA damage, increased SA-β-gal activity, and increased expression of aging-related factors p16 and p21 during differentiation. Moreover, aged myoblasts showed a reduction in the expression of MRFs and metabolic/anabolic factors, along with an increased expression of apoptotic, atrophy and inflammatory factors. A diminished differentiation capacity characterized the aged myoblasts which, in combination with the induction of apoptotic and atrophy factors, indicated a disrupted myogenic lineage in the senescent muscle cells.

Lactobacillus plantarum HY7715 Ameliorates Sarcopenia by Improving Skeletal Muscle Mass and Function in Aged Balb/c Mice

“…The C2C12 cell line derived from murine skeletal muscle is an established model used to investigate muscle differentiation [ 33 , 34 , 35 ]. To identify how HY7715 induces muscle myogenic activation, C2C12 cells were differentiated for 5 days, and HY7715 was added to the differentiation media for the last 48 h. Whole-cell extracts of HY7715 were lysed, and the cytoplasmic and membrane fractions were isolated.…”

Diverse Roles of Cellular Senescence in Skeletal Muscle Inflammation, Regeneration, and Therapeutics

“…In humans, skeletal muscle tissue from older women (65–71 years old) expresses higher levels of CDKN1A than skeletal tissue from young women (20–29 years old) ( Welle et al, 2004 ). Several potential mechanisms of muscle cell senescence–mediated, aging-associated muscle loss (sarcopenia) were proposed based on an in vitro study using C2C12 cells, a murine myoblast cell line ( Alcalde-Estévez et al, 2020 ; Mijares et al, 2021 ; Moustogiannis et al, 2021 ). These studies revealed that elevated intracellular Ca 2+ concentration or endothelin-1 receptor in C2C12 cells induces cellular senescence, and senescent C2C12 cells increase the expression of apoptotic, atrophic, and inflammatory factors, which might in turn induce sarcopenia mediated by muscle cell senescence ( Alcalde-Estévez et al, 2020 ; Mijares et al, 2021 ; Moustogiannis et al, 2021 ).…”

“…Several potential mechanisms of muscle cell senescence–mediated, aging-associated muscle loss (sarcopenia) were proposed based on an in vitro study using C2C12 cells, a murine myoblast cell line ( Alcalde-Estévez et al, 2020 ; Mijares et al, 2021 ; Moustogiannis et al, 2021 ). These studies revealed that elevated intracellular Ca 2+ concentration or endothelin-1 receptor in C2C12 cells induces cellular senescence, and senescent C2C12 cells increase the expression of apoptotic, atrophic, and inflammatory factors, which might in turn induce sarcopenia mediated by muscle cell senescence ( Alcalde-Estévez et al, 2020 ; Mijares et al, 2021 ; Moustogiannis et al, 2021 ). On the other hand, recent in vivo analysis of human skeletal muscle revealed that there is no difference in the abundance of γH2AX-positive myonuclei between young individuals (21–30 years old) and old individuals (70–86 years old) ( Dungan et al, 2020 ).…”

The Effects of Mechanical Loading Variations on the Hypertrophic, Anti-Apoptotic, and Anti-Inflammatory Responses of Differentiated Cardiomyocyte-like H9C2 Cells

“…On the other hand, mechanical loading can have beneficial or detrimental effects on differentiated cardiomyocytes, depending on its specific characteristics [29], by activating pro-inflammatory factors, such as IL-1β, TNF-α, IL-6, and NF-kB, as well as muscle atrophy and pro-apoptotic factors, such as myostatin, muscle-specific ubiquitin ligase Atrogin-1 (MaFbx), FoxO1, and p53, which have been negatively implicated in cell growth and survival, inducing cardiac muscle wasting and promoting heart failure [29][30][31][32][33][34][35][36].…”