Three-dimensional niche stiffness synergizes with Wnt7a to modulate the extent of satellite cell symmetric self-renewal divisions

Mol Biol Cell. 2020 Jul 21;31(16):1703-1713. doi: 10.1091/mbc.E20-01-0078. Epub 2020 Jun 3.

Abstract

Satellite cells (SCs), the resident adult stem cells of skeletal muscle, are required for tissue repair throughout life. While many signaling pathways are known to control SC self-renewal, less is known about the mechanisms underlying the spatiotemporal control of self-renewal during skeletal muscle repair. Here, we measured biomechanical changes that accompany skeletal muscle regeneration and determined the implications on SC fate. Using atomic force microscopy, we quantified a 2.9-fold stiffening of the SC niche at time-points associated with planar-oriented symmetric self-renewal divisions. Immunohistochemical analysis confirms increased extracellular matrix deposition within the basal lamina. To test whether three-dimensional (3D) niche stiffness can alter SC behavior or fate, we embedded isolated SC-associated muscle fibers within biochemically inert agarose gels tuned to mimic native tissue stiffness. Time-lapse microscopy revealed that a stiff 3D niche significantly increased the proportion of planar-oriented divisions, without effecting SC viability, fibronectin deposition, or fate change. We then found that 3D niche stiffness synergizes with WNT7a, a biomolecule shown to control SC symmetric self-renewal divisions via the noncanonical WNT/planar cell polarity pathway, to modify stem cell pool expansion. Our results provide new insights into the role of 3D niche biomechanics in regulating SC fate choice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells
  • Animals
  • Cell Differentiation / physiology
  • Cell Proliferation / physiology
  • Elasticity / physiology
  • Extracellular Matrix / metabolism
  • Female
  • Fibronectins / genetics
  • Fibronectins / metabolism
  • Hardness / physiology
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Atomic Force / methods
  • Muscle Fibers, Skeletal
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / physiology*
  • Regeneration / physiology
  • Satellite Cells, Skeletal Muscle / metabolism*
  • Satellite Cells, Skeletal Muscle / physiology
  • Signal Transduction / physiology
  • Stem Cell Niche / physiology
  • Wnt Proteins / genetics
  • Wnt Proteins / metabolism
  • Wound Healing / physiology*

Substances

  • FN1 protein, human
  • Fibronectins
  • WNT7A protein, human
  • Wnt Proteins