Non–fibro-adipogenic pericytes from human embryonic stem cells attenuate degeneration of the chronically injured mouse muscle
Gina M. Mosich, … , Frank A. Petrigliano, Ayelet Dar
Gina M. Mosich, … , Frank A. Petrigliano, Ayelet Dar
Published December 19, 2019
Citation Information: JCI Insight. 2019;4(24):e125334. https://doi.org/10.1172/jci.insight.125334.
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Research Article Muscle biology Stem cells

Non–fibro-adipogenic pericytes from human embryonic stem cells attenuate degeneration of the chronically injured mouse muscle

Abstract

Massive tears of the rotator cuff (RC) are associated with chronic muscle degeneration due to fibrosis, fatty infiltration, and muscle atrophy. The microenvironment of diseased muscle often impairs efficient engraftment and regenerative activity of transplanted myogenic precursors. Accumulating myofibroblasts and fat cells disrupt the muscle stem cell niche and myogenic cell signaling and deposit excess disorganized connective tissue. Therefore, restoration of the damaged stromal niche with non–fibro-adipogenic cells is a prerequisite to successful repair of an injured RC. We generated from human embryonic stem cells (hES) a potentially novel subset of PDGFR-β+CD146+CD34–CD56– pericytes that lack expression of the fibro-adipogenic cell marker PDGFR-α. Accordingly, the PDGFR-β+PDGFR-α– phenotype typified non–fibro-adipogenic, non-myogenic, pericyte-like derivatives that maintained non–fibro-adipogenic properties when transplanted into chronically injured murine RCs. Although administered hES pericytes inhibited developing fibrosis at early and late stages of progressive muscle degeneration, transplanted PDGFR-β+PDGFR-α+ human muscle-derived fibro-adipogenic progenitors contributed to adipogenesis and greater fibrosis. Additionally, transplanted hES pericytes substantially attenuated muscle atrophy at all tested injection time points after injury. Coinciding with this observation, conditioned medium from cultured hES pericytes rescued atrophic myotubes in vitro. These findings imply that non–fibro-adipogenic hES pericytes recapitulate the myogenic stromal niche and may be used to improve cell-based treatments for chronic muscle disorders.

Authors

Gina M. Mosich, Regina Husman, Paras Shah, Abhinav Sharma, Kevin Rezzadeh, Temidayo Aderibigbe, Vivian J. Hu, Daniel J. McClintick, Genbin Wu, Jonathan D. Gatto, Haibin Xi, April D. Pyle, Bruno Péault, Frank A. Petrigliano, Ayelet Dar

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Figure 2

Human embryonic stem cell derivatives are lineage-restricted pericytes.

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Human embryonic stem cell derivatives are lineage-restricted pericytes. ...
(A) Representative flow cytometry dot plot of CD34, CD45, and CD146 expression by hES cell derivatives expanded in EGM-2 medium at passage 0. Flow cytometry analysis of expression of characteristic cell surface markers: PDGFR-β, CD73, and CD146 (pericytic); CD45 (hematopoietic); and CD34 (endothelial, hematopoietic progenitors and adventitial cells) by expanded hES derivatives at passages 2 and 9. (B) Vasculogenic features of CD146+ PC-like cells from hES are demonstrated by tube formation on Matrigel. (C) Representative dot plots of flow cytometry analysis of CD34, PDGFR-α, CD146, and CD56 expression by expanded hES pericytic derivatives. (D–K) hES PCs lack fibro-adipogenic differentiation capability. Adipocytes are not detected in control (D) and adipogenic cultures (E). Negative Oil Red O staining of non-induced control (F) and adipogenic induced cultures (G). Representative images of picrosirius red staining for collagen of control (H) and TGF-β1–treated (I) hES PC cultures. Spectrophotometric quantification of collagen production (mean ± SEM) by non-induced and TGF-β1–induced hES PC cultures at the indicated time points (J). #P < 0.00001 compared with control and TGF-β1 induced hES PCs that were cultured for 4 days and 2 weeks. *P < 0.00001 compared with control and TGF-β1–induced hES PCs that were cultured for 4 days (1-way ANOVA). TGF-β1 does not induce the expression of α-smooth muscle actin (K, α-SMA in green, nuclear staining for DAPI in blue) by hES PCs. (L and M) Poor staining for alizarin red demonstrates limited osteogenic differentiation of induced hES PCs. (N) Sorting strategy based on the expression of CD146 and CD56 by human muscle cells expanded in EGM-2 medium at passage 0. Flow cytometry analysis of the expression of PDGFR-β, PDGFR-α, and CD45 by sorted CD56– cells at passages 1–2 (right). (O–R) Myogenic (O and P) and adipogenic (Q and R) cultures of PDGFR-α+CD56– (O and Q) and PDGFR-α–CD56+ (P and R) cells. (S) Concentration of collagen in control and TGF-β1–induced PDGFR-α+CD56– cell cultures (mean ± SEM). Data were pooled from 3 independent experiments (n = 3 donors) with triplicates. *P < 0.005 compared with untreated cultures (1-way ANOVA). Scale bars: 100 μm.