Scleroderma Mesenchymal Stem Cells display a different phenotype from healthy controls; implications for regenerative medicine

Angiogenesis. 2013 Jul;16(3):595-607. doi: 10.1007/s10456-013-9338-9. Epub 2013 Feb 15.

Abstract

Introduction: Vascular involvement is a key feature of Systemic sclerosis (SSc). Although the pericytes/endothelial cells (ECs) cross-talk regulates vessels formation, no evidences about the pericytes contribution to ineffective angiogenesis in SSc are available. Recent findings showed similarities between pericytes and Bone Marrow Mesenchymal Stem Cells (BM-MSCs). Due to difficulties in pericytes isolation, this work explores the possibility to use BM-MSCs as pericytes surrogate, clarifying their role in supporting neo-angiogenesis during SSc.

Methods: To demonstrate their potential to normally differentiate into pericytes, both SSc and healthy controls (HC) BM-MSCs were treated with TGF-β and PDGF-BB. The expression of pericytes specific markers (α-SMA, NG2, RGS5 and desmin) was assessed by qPCR, western blot, and immunofluorescence; chemioinvasion and capillary morphogenesis were also performed. Cell-sorting of BM-MSCs co-cultured with HC-ECs was used to identify a possible change in contractile proteins genes expression.

Results: We showed that BM-MSCs isolated from SSc patients displayed an up-regulation of α-SMA and SM22α genes and a reduced proliferative activity. Moreover during SSc, both TGF-β and PDGF-BB can specifically modulate BM-MSCs toward pericytes. TGF-β was found interfering with the PDGF-BB effects. Using BM-MSCs/MVECs co-culture system we observed that SSc BM-MSCs improve ECs tube formation in stressed condition, and BM-MSCs, sorted after co-culture, showed a reduced α-SMA and SM22α gene expression.

Conclusions: BM-MSCs from SSc patients behave as pericytes. They display a more mature and myofibroblast-like phenotype, probably related to microenvironmental cues operating during the disease. After their co-culture with HC-MVECs, SSc BM-MSCs underwent to a phenotypic modulation which re-programs these cells toward a pro-angiogenic behaviour.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Becaplermin
  • Blotting, Western
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology
  • Cells, Cultured
  • DNA Primers / genetics
  • Dose-Response Relationship, Drug
  • Endothelium, Vascular / cytology*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Humans
  • Mesenchymal Stem Cells / physiology*
  • Microfilament Proteins / metabolism
  • Muscle Proteins / metabolism
  • Neovascularization, Physiologic / physiology*
  • Pericytes / physiology
  • Phenotype*
  • Proto-Oncogene Proteins c-sis / pharmacology
  • Real-Time Polymerase Chain Reaction
  • Regenerative Medicine / methods*
  • Scleroderma, Systemic / physiopathology*
  • Scleroderma, Systemic / therapy
  • Transforming Growth Factor beta / pharmacology

Substances

  • ACTA2 protein, human
  • Actins
  • DNA Primers
  • Microfilament Proteins
  • Muscle Proteins
  • Proto-Oncogene Proteins c-sis
  • Transforming Growth Factor beta
  • transgelin
  • Becaplermin