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Isolation of Perivascular Multipotent Precursor Cell Populations from Human Cardiac Tissue
Published on: October 8, 2016
Human myocardial pericytes: multipotent mesodermal precursors exhibiting cardiac specificity
William C W Chen1, James E Baily, Mirko Corselli
1Department of Bioengineering, University of Pittsburgh, Pennsylvania, USA; Department of Orthopedic Surgery, University of Pittsburgh, Pennsylvania, USA; Stem Cell Research Centre, University of Pittsburgh, Pennsylvania, USA.
Abstract:
Perivascular mesenchymal precursor cells (i.e., pericytes) reside in skeletal muscle where they contribute to myofiber regeneration; however, the existence of similar microvessel-associated regenerative precursor cells in cardiac muscle has not yet been documented. We tested whether microvascular pericytes within human myocardium exhibit phenotypes and multipotency similar to their anatomically and developmentally distinct counterparts. Fetal and adult human heart pericytes (hHPs) express canonical pericyte markers in situ, including CD146, NG2, platelet-derived growth factor receptor (PDGFR) β, PDGFRα, alpha-smooth muscle actin, and smooth muscle myosin heavy chain, but not CD117, CD133, and desmin, nor endothelial cell (EC) markers. hHPs were prospectively purified to homogeneity from ventricular myocardium by flow cytometry, based on a combination of positive- (CD146) and negative-selection (CD34, CD45, CD56, and CD117) cell lineage markers. Purified hHPs expanded in vitro were phenotypically similar to human skeletal muscle-derived pericytes (hSkMPs). hHPs express mesenchymal stem/stromal cell markers in situ and exhibited osteo-, chondro-, and adipogenic potentials but, importantly, no ability for skeletal myogenesis, diverging from pericytes of all other origins. hHPs supported network formation with/without ECs in Matrigel cultures; hHPs further stimulated angiogenic responses under hypoxia, markedly different from hSkMPs. The cardiomyogenic potential of hHPs was examined following 5-azacytidine treatment and neonatal cardiomyocyte coculture in vitro, and intramyocardial transplantation in vivo. Results indicated cardiomyocytic differentiation in a small fraction of hHPs. In conclusion, human myocardial pericytes share certain phenotypic and developmental similarities with their skeletal muscle homologs, yet exhibit different antigenic, myogenic, and angiogenic properties. This is the first example of an anatomical restriction in the developmental potential of pericytes as native mesenchymal stem cells.
Insights
Human heart pericytes (hHPs) share traits with skeletal muscle pericytes but have distinct myogenic and angiogenic properties. This study reveals anatomical restrictions in pericyte stem cell potential.
Area of Science:
- Cardiovascular Biology
- Stem Cell Biology
- Regenerative Medicine
Background:
- Perivascular mesenchymal precursor cells (pericytes) aid skeletal muscle regeneration.
- The presence and function of cardiac pericytes as regenerative precursors remain largely undocumented.
Purpose of the Study:
- To investigate if human heart pericytes (hHPs) exhibit similar phenotypes and multipotency to other pericyte types.
- To characterize the differentiation potential and functional properties of hHPs.
Main Methods:
- Prospective purification of hHPs from human myocardium via flow cytometry using specific cell surface markers.
- In vitro expansion and characterization of hHPs for mesenchymal stem/stromal cell marker expression and differentiation potential (osteogenesis, chondrogenesis, adipogenesis).
- Assessment of hHPs' angiogenic capacity and cardiomyogenic potential through in vitro assays and in vivo transplantation.
Main Results:
- Purified hHPs expressed canonical pericyte markers and mesenchymal stem/stromal cell markers.
- hHPs demonstrated osteo-, chondro-, and adipogenic potential but lacked skeletal myogenesis capacity.
- hHPs supported endothelial cell network formation, stimulated angiogenesis under hypoxia, and showed limited cardiomyocytic differentiation.
Conclusions:
- Human myocardial pericytes share some similarities with skeletal muscle pericytes but possess unique antigenic, myogenic, and angiogenic properties.
- This study presents the first evidence of anatomical restriction in the developmental potential of pericytes as native mesenchymal stem cells.
- Findings highlight distinct functional roles of pericytes based on their anatomical location.
