PMID- 33543006
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210206
IS  - 2590-0285 (Electronic)
IS  - 2590-0285 (Linking)
VI  - 2
DP  - 2019 May
TI  - Adherent muscle connective tissue fibroblasts are phenotypically and 
      biochemically equivalent to stromal fibro/adipogenic progenitors.
PG  - 100006
LID - 10.1016/j.mbplus.2019.04.003 [doi]
LID - 100006
AB  - Extracellular matrix (ECM) gives structure, support, and is the niche for several 
      cells found in skeletal muscle. ECM is mainly produced by muscle connective 
      tissue (CT) fibroblasts during development and regeneration. Stromal 
      fibroadipogenic progenitors (FAPs) are CT fibroblasts-like mesenchymal 
      progenitors (MPs) with important roles in regeneration and degeneration. Chronic 
      damage restrains the normal regenerative behavior of muscle fibroblasts/FAPs. 
      Thus, the isolation and study of these mesenchymal progenitors are of crucial 
      importance for understanding their behavior and biology. We investigated whether 
      adult muscle CT fibroblasts (hereafter referred to as adherent fibroblasts 
      [aFbs]) cultured via pre-plating strategy belong to a heterogeneous population of 
      FAPs. By combining microscopy, western blot analyses, flow cytometry, and FACS we 
      determined that aFbs isolated from skeletal muscle largely overlap with FAPs. In 
      addition, we used the PDGFRalpha(EGFP) mice in order to corroborate our results with 
      EGFP(+) FAPs. Moreover, our strategy allows the isolation of activated EGFP(+) 
      FAPs from the murine DMD model PDGFRalpha(EGFP); mdx and PDGFRalpha(EGFP) denervated 
      mice. Here we report that 1 h 30 min of pre-plating strategy allows the isolation 
      and culture of a highly enriched population of aFbs. These cells are 
      phenotypically and biochemically a FAPs-like population of adherent cells. In 
      addition, aFbs respond in the same fashion as FAPs to Nilotinib, an inducer of 
      FAPs apoptosis. Moreover, flow cytometry characterization of these aFbs suggests 
      that 85% of them express the MP marker PDGFRalpha, and isolation of aFbs from the 
      PDGFRalpha(EGFP) mice suggests that 75% of them show high EGFP expression. 
      Furthermore, TGF-beta1 induces aFbs proliferation, myofibroblast differentiation, 
      and ECM production. We were also able to isolate activated aFbs from skeletal 
      muscle of the DMD mice and from the PDGFRalpha(EGFP) mice 2-days after denervation. 
      Our findings suggest that the in vitro pre-plating strategy allows the isolation 
      and culture of a relatively pure aFbs population, which resembles FAPs in vitro.
CI  - (c) 2019 The Authors.
FAU - Contreras, Osvaldo
AU  - Contreras O
AD  - Departamento de Biologia Celular y Molecular and Center for Aging and 
      Regeneration (CARE-ChileUC), Facultad de Ciencias Biologicas, Pontificia 
      Universidad Catolica de Chile, Santiago, Chile.
AD  - Biomedical Research Centre, Department of Medical Genetics, 2222 Health Sciences 
      Mall, University of British Columbia, Vancouver, BC, Canada.
FAU - Rossi, Fabio M
AU  - Rossi FM
AD  - Biomedical Research Centre, Department of Medical Genetics, 2222 Health Sciences 
      Mall, University of British Columbia, Vancouver, BC, Canada.
FAU - Brandan, Enrique
AU  - Brandan E
AD  - Departamento de Biologia Celular y Molecular and Center for Aging and 
      Regeneration (CARE-ChileUC), Facultad de Ciencias Biologicas, Pontificia 
      Universidad Catolica de Chile, Santiago, Chile.
LA  - eng
PT  - Journal Article
DEP - 20190417
PL  - Netherlands
TA  - Matrix Biol Plus
JT  - Matrix biology plus
JID - 101775320
PMC - PMC7852197
OTO - NOTNLM
OT  - Adherent Fibroblasts, aFbs
OT  - Connective tissue, CT
OT  - Extracellular matrix, ECM
OT  - FAPs
OT  - Fibroadipogenic progenitors, FAPs
OT  - Fibrosis
OT  - Fluorescence-activated cell sorting, FACS
OT  - Mesenchymal Progenitors, MPs
OT  - Mesenchymal progenitors
OT  - Muscle stem cells, MuSCs
OT  - PDGFRalpha
OT  - Platelet-derived growth factor receptor alpha, PDGFRalpha
OT  - Skeletal muscle
OT  - TGF-beta signaling
OT  - Transcription factor, TF
OT  - Transforming growth factor type-beta, (TGF-beta)
EDAT- 2019/04/17 00:00
MHDA- 2019/04/17 00:01
PMCR- 2019/04/17
CRDT- 2021/02/05 06:00
PHST- 2019/02/21 00:00 [received]
PHST- 2019/04/12 00:00 [revised]
PHST- 2019/04/12 00:00 [accepted]
PHST- 2021/02/05 06:00 [entrez]
PHST- 2019/04/17 00:00 [pubmed]
PHST- 2019/04/17 00:01 [medline]
PHST- 2019/04/17 00:00 [pmc-release]
AID - S2590-0285(19)30005-5 [pii]
AID - 100006 [pii]
AID - 10.1016/j.mbplus.2019.04.003 [doi]
PST - epublish
SO  - Matrix Biol Plus. 2019 Apr 17;2:100006. doi: 10.1016/j.mbplus.2019.04.003. 
      eCollection 2019 May.