Fibroblast rejuvenation by mechanical reprogramming and redifferentiation

Proc Natl Acad Sci U S A. 2020 May 12;117(19):10131-10141. doi: 10.1073/pnas.1911497117. Epub 2020 Apr 29.


Over the course of the aging process, fibroblasts lose contractility, leading to reduced connective-tissue stiffness. A promising therapeutic avenue for functional rejuvenation of connective tissue is reprogrammed fibroblast replacement, although major hurdles still remain. Toward this, we recently demonstrated that the laterally confined growth of fibroblasts on micropatterned substrates induces stem-cell-like spheroids. In this study, we embedded these partially reprogrammed spheroids in collagen-I matrices of varying densities, mimicking different three-dimensional (3D) tissue constraints. In response to such matrix constraints, these spheroids regained their fibroblastic properties and sprouted to form 3D connective-tissue networks. Interestingly, we found that these differentiated fibroblasts exhibit reduced DNA damage, enhanced cytoskeletal gene expression, and actomyosin contractility. In addition, the rejuvenated fibroblasts show increased matrix protein (fibronectin and laminin) deposition and collagen remodeling compared to the parental fibroblast tissue network. Furthermore, we show that the partially reprogrammed cells have comparatively open chromatin compaction states and may be more poised to redifferentiate into contractile fibroblasts in 3D-collagen matrix. Collectively, our results highlight efficient fibroblast rejuvenation through laterally confined reprogramming, which has important implications in regenerative medicine.

Keywords: engineered 3D tissue; lateral confinement; redifferentiation; rejuvenation; stem-cell-like state.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • Cell Differentiation*
  • Cell Proliferation
  • Cells, Cultured
  • Cellular Reprogramming*
  • Collagen Type I / metabolism
  • Extracellular Matrix / metabolism
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism
  • Fibronectins / metabolism
  • Humans
  • Laminin / metabolism
  • Mice
  • NIH 3T3 Cells
  • Regenerative Medicine*
  • Rejuvenation / physiology*
  • Spheroids, Cellular / cytology
  • Spheroids, Cellular / metabolism


  • Collagen Type I
  • Fibronectins
  • Laminin