Quiescence of human muscle stem cells is favored by culture on natural biopolymeric films

Abstract

BackgroundSatellite cells are quiescent resident muscle stem cells that present an important potential to regenerate damaged tissue. However, this potential is diminished once they are removed from their niche environment in vivo, prohibiting the long-term study and genetic investigation of these cells. This study therefore aimed to provide a novel biomaterial platform for the in-vitro culture of human satellite cells that maintains their stem-like quiescent state, an important step for cell therapeutic studies.MethodsHuman muscle satellite cells were isolated from two donors and cultured on soft biopolymeric films of controlled stiffness. Cell adhesive phenotype, maintenance of satellite cell quiescence and capacity for gene manipulation were investigated using FACS, western blotting, fluorescence microscopy and electron microscopy.ResultsAbout 85% of satellite cells cultured in vitro on soft biopolymer films for 3 days maintained expression of the quiescence marker Pax7, as compared with 60% on stiffer films and 50% on tissue culture plastic. The soft biopolymeric films allowed satellite cell culture for up to 6 days without renewing the media. These cells retained their stem-like properties, as evidenced by the expression of stem cell markers and reduced expression of differentiated markers. In addition, 95% of cells grown on these soft biopolymeric films were in the G0/G1 stage of the cell cycle, as opposed to those grown on plastic that became activated and began to proliferate and differentiate.ConclusionsOur study identifies a new biomaterial made of a biopolymer thin film for the maintenance of the quiescence state of muscle satellite cells. These cells could be activated at any point simply by replating them onto a plastic culture dish. Furthermore, these cells could be genetically manipulated by viral transduction, showing that this biomaterial may be further used for therapeutic strategies.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0556-8) contains supplementary material, which is available to authorized users.

Dystrophin Expressing Chimeric (DEC) Human Cells Provide a Potential Therapy for Duchenne Muscular Dystrophy

“…Furthermore, low expression (below 2%) of CD45, the hematopoietic stem cell marker, combined with high expression of CD56 (85%), the myoblast specific marker, confirms high myogenic potential of DEC. The efficacy of DEC therapy is cell-passage dependent and the optimal MBs passage for ex vivo fusion ranges between passage 5 and 7, since at passage 4 cells do not reach the required number and after passage 7 differentiation potential of MBs decreases [ 31 , 32 ]. The genotype stability of both DEC lines was preserved after fusion as confirmed by STR-PCR and PCR-rSSOP profiling.…”

Skeletal muscle stem cells in comfort and stress

“…32 Efforts have also been made to artificially recapitulate cellular quiescence or the niche environment ex vivo. 33–35…”

Abnormalities in Skeletal Muscle Myogenesis, Growth, and Regeneration in Myotonic Dystrophy

“…Novel strategies for satellite cell culture and preservation of self-renewal capacity before transplantation into muscle have now become available. Cell culture on pliable soft hydrogel matrices, in combination with pharmacological inhibition of p38/MAPK signaling ( 277 ) or culture on natural biopolymeric films ( 278 ) simulate the conditions of the muscle stem cell niche and help to preserve MuSC quiescence and enhance their self-renewal capacity. Also modulation of PAX7 expression may thereby be of help ( 279 ).…”