Murine myoblast migration: influence of replicative ageing and nutrition

Abstract

Cell migration is central to skeletal muscle repair following damage. Leucine and b-Hydroxy bmethylbutyric acid (HMB) are supplements consumed for recovery from muscle damaging exercise in humans, however, their impact on muscle cell migration with age is not yet understood. We hypothesised that replicatively aged (''aged''; P46-P48) myoblasts would be less efficient at basal and supplemented repair versus parental controls (''control''; P12-P16). Aged and control myoblasts were scratch-damaged and migration velocity, directionality and distance assessed over 48 h in the absence and presence of leucine (10 mM) or HMB (10 mM) ± PI3K/Akt (LY294002 10 lM), ERK (PD98059 5 lM) or mTOR (rapamycin 0.5 lM) inhibition. Opposing our hypothesis, aged cells displayed increased velocities, directionality and distance migrated (P \ 0.001) versus control. Leucine and HMB significantly increased (P \ 0.001) the same parameters in control cells. The supplements were with smaller, albeit significant impact on aged cell velocity (P \ 0.001) and in the presence of HMB only, distance (P = 0.041). Inhibitor studies revealed that, PI3K and ERK activation were essential for velocity, directionality and migration distance of aged cells in basal conditions, whereas mTOR was important for directionality only. While PI3K activation was critical for all parameters in control cells (P \ 0.001), inhibition of ERK or mTOR improved, rather than reduced, control cell migration distance. Enhanced basal velocity, directionality and distance in aged cells required ERK and PI3K activation. By contrast, in control cells, basal migration was underpinned by PI3K activation, and facilitated by leucine or HMB supplementation, to migration levels seen in aged cells. These data suggest that replicatively aged myoblasts are not anabolically resistant per se, but are capable of efficient repair, underpinned by altered signaling pathways, compared with unaged control myoblasts.

Cellular dynamics of myogenic cell migration: molecular mechanisms and implications for skeletal muscle cell therapies

“…(Chal et al, 2016;Hicks et al, 2018)) could generate innovative advanced therapy medicinal products (ATMPs) with controllable proliferation, migration and differentiation capacity. 2 Phenotypic disparities between young and aged myogenic cell types have been investigated (Collins-Hooper et al, 2012;Brown et al, 2017;Rotini et al, 2018). In addition to regenerative capacity, migration has been suggested to be altered with ageing: studying and understanding migration dynamics in young vs aged myogenic cells may highlight specific pathways to modulate to enhance migration of ATMPs.…”

“…(Tedesco et al , 2012; Darabi et al , 2012)) or transgene‐free protocols (e.g. (Chal et al , 2016; Hicks et al , 2018)) could generate innovative advanced therapy medicinal products (ATMPs) with controllable proliferation, migration and differentiation capacity. Phenotypic disparities between young and aged myogenic cell types have been investigated (Collins‐Hooper et al , 2012; Brown et al , 2017; Rotini et al , 2018). In addition to regenerative capacity, migration has been suggested to be altered with ageing: studying and understanding migration dynamics in young vs aged myogenic cells may highlight specific pathways to modulate to enhance migration of ATMPs. Lastly, omics‐based comparative studies of myogenic cell populations treated with pro‐migratory factors or small molecules may unravel druggable pathways which could be further modulated to enhance cell motility.…”

CCR3 antagonist protects against induced cellular senescence and promotes rejuvenation in periodontal ligament cells for stimulating pulp regeneration in the aged dog

“…Numerous cytokines, molecular pathways, and chemokines involved in persistent chronic inflammation induce direct tissue degeneration and cause multiple age-related diseases 36 . Regeneration in aged tissues is declined due to resident stem cell senescence 32,37 , chronic inflammation 38 , decreased migration activity 39 . Angiogenesis/vaculogenesis 40 and neurogenesis 41 are also decreased.…”

Degradation of ribosome and chaperone proteins is attenuated during the Differentiation of Replicatively Aged C2C12 Myoblasts

“…(2) replicatively aged (high passage 48 -50). The aged C2C12 myoblasts were based on a previous model of replicative aging [11,12]. In brief, the aged myoblasts underwent 140 -150 population doublings.…”

“…Parabiotic studies conducted in vivo [7] and in vitro [8,9] have established an important role of the host environment on muscle regeneration and satellite cell function, whilst replicative aging of cells in vitro has demonstrated age-related impairments in differentiation capacity [10] that occur in the absence of the confounding effects of changes in systemic environment. Replicatively aged C2C12 myoblasts exhibit impaired differentiation and fusion [11,12] similar to human replicatively-aged primary myoblasts [13] and cells and tissue isolated from older rodents and humans [13][14][15][16][17][18]. Myoblast differentiation is orchestrated by complex programmes of cell signalling [19] and altered gene expression [17] that result in dynamic changes to the cell proteome during differentiation [20,21].…”