Angiotensin II and TGF-β1 Induce Alterations in Human Amniotic Fluid-Derived Mesenchymal Stem Cells Leading to Cardiomyogenic Differentiation Initiation
- Author:
Monika GASIŪNIENĖ
1
;
Gintautas PETKUS
;
Dalius MATUZEVIČIUS
;
Dalius NAVAKAUSKAS
;
Rūta NAVAKAUSKIENĖ
Author Information
- Publication Type:Original Article
- Keywords: Myocytes; Cardiac; Amniotic fluid; Cell differentiation; Chromatin; Histones
- MeSH: Amniocentesis; Amniotic Fluid; Angiotensin II; Angiotensins; Blotting, Western; Calcium; Cell Differentiation; Cell- and Tissue-Based Therapy; Chromatin; Chromatin Assembly and Disassembly; Connexin 43; Epigenomics; Female; Fluorescent Antibody Technique; Gap Junctions; Glycolysis; Histones; Humans; Ion Channels; Mesenchymal Stromal Cells; Muscle Cells; Oxidative Phosphorylation; Phenotype; Polycomb Repressive Complex 1; Pregnant Women; Smegmamorpha; Stem Cells; Tissue Engineering; Transcription Factors; Up-Regulation
- From:International Journal of Stem Cells 2019;12(2):251-264
- CountryRepublic of Korea
- Language:English
- Abstract: BACKGROUND AND OBJECTIVES: Human amniotic fluid-derived mesenchymal stem cells (AF-MSCs) may be a valuable source for cardiovascular tissue engineering and cell therapy. The aim of this study is to verify angiotensin II and transforming growth factor-beta 1 (TGF-β1) as potential cardiomyogenic differentiation inducers of AF-MSCs. METHODS AND RESULTS: AF-MSCs were obtained from amniocentesis samples from second-trimester pregnant women, isolated and characterized by the expression of cell surface markers (CD44, CD90, CD105 positive; CD34 negative) and pluripotency genes (OCT4, SOX2, NANOG, REX1). Cardiomyogenic differentiation was induced using different concentrations of angiotensin II and TGF-β1. Successful initiation of differentiation was confirmed by alterations in cell morphology, upregulation of cardiac genes-markers NKX2-5, TBX5, GATA4, MYH6, TNNT2, DES and main cardiac ion channels genes (sodium, calcium, potassium) as determined by RT-qPCR. Western blot and immunofluorescence analysis revealed the increased expression of Connexin43, the main component of gap junctions, and Nkx2.5, the early cardiac transcription factor. Induced AF-MSCs switched their phenotype towards more energetic and started utilizing oxidative phosphorylation more than glycolysis for energy production as assessed using Agilent Seahorse XF analyzer. The immune analysis of chromatin-modifying enzymes DNMT1, HDAC1/2 and Polycomb repressive complex 1 and 2 (PRC1/2) proteins BMI1, EZH2 and SUZ12 as well as of modified histones H3 and H4 indicated global chromatin remodeling during the induced differentiation. CONCLUSIONS: Angiotensin II and TGF-β1 are efficient cardiomyogenic inducers of human AF-MSCs; they initiate alterations at the gene and protein expression, metabolic and epigenetic levels in stem cells leading towards cardiomyocyte-like phenotype formation.
