RT Journal Article
SR Electronic
T1 Bioengineered human skeletal muscle with a Pax7+ satellite cell niche capable of functional regeneration
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.05.04.076828
DO 10.1101/2020.05.04.076828
A1 J.W. Fleming
A1 A.J. Capel
A1 R.P. Rimington
A1 P. Wheeler
A1 O.G. Davies
A1 M.P. Lewis
YR 2020
UL http://biorxiv.org/content/early/2020/05/07/2020.05.04.076828.abstract
AB Skeletal muscle (SkM) regenerates following injury, replacing damaged tissue with high fidelity. However, in serious injuries non-regenerative defects leave patients with loss of function, increased re-injury risk and often chronic pain. Progress in treating these non-regenerative defects has been slow, with advances only occurring where a comprehensive understanding of regeneration has been gained. Tissue engineering has allowed the development of bioengineered models of SkM which regenerate following injury to support research in regenerative physiology. To date however, no studies have utilised human myogenic precursor cells (hMPCs) to closely mimic human physiology due to difficulties generating sufficient cell numbers and the relatively low myogenic potential of hMPCs. Here we address problems associated with cell number and hMPC mitogenicity using magnetic association cell sorting (MACS), for the marker CD56, and media supplementation with fibroblast growth factor 2 (FGF-2) and B-27 supplement. Cell sorting allowed extended expansion of myogenic cells and supplementation was shown to improve myogenesis within engineered tissues and force generation at maturity. In addition, these engineered human SkM contained a Pax7+ niche and regenerated following Barium Chloride (BaCl2) injury. Following injury, reductions in function (87.5%) and myotube number (33.3%) were observed, followed by a proliferative phase with increased MyoD+ cells and a subsequent recovery of function and myotube number. An expansion of the Pax7+ cell population was observed across recovery suggesting an ability to generate Pax7+ cells within the tissue, similar to the self-renewal of satellite cells seen in vivo. This work outlines an engineered human SkM capable of functional regeneration following injury, built upon an open source system adding to the pre-clinical testing toolbox to improve the understanding of basic regenerative physiology.Competing Interest StatementThe authors have declared no competing interest.GMGrowth mediumDMDifferentiation mediumECMExtracellular matrixMPCMyogenic precursor cellhMPCHuman myogenic precursor cellMyHCMyosin heavy chainSCSatellite cellVMLVolumetric muscle lossFAPFibro/adipogenic cell3DThree dimensionalPSCPluripotent stem cellDMEMDulbecco’s Modified Eagle’s MediumFBSFetal Bovine SerumP/SPenicillin/StreptomycinMEMMinimal essential mediumPBSPhosphate buffered salineKRHKrebs-Ringer-HEPESBaCl2Barium Chloride