Multipotent Vasculogenic Pericytes From Human Pluripotent Stem Cells Promote Recovery of Murine Ischemic Limb

Abstract

Background-Pericytes represent a unique subtype of microvessel-residing perivascular cells with diverse angiogenic functions and multilineage developmental features of mesenchymal stem cells. Although various protocols for derivation of endothelial and/or smooth muscle cells from human pluripotent stem cells (hPSC, either embryonic or induced) have been described, the emergence of pericytes in the course of hPSC maturation has not yet been elucidated. Methods and Results-We found that during hPSC development, spontaneously differentiating embryoid bodies give rise to CD105 ϩ CD90 ϩ CD73 ϩ CD31 Ϫ multipotent clonogenic mesodermal precursors, which can be isolated and efficiently expanded. Isolated and propagated cells expressed characteristic pericytic markers, including CD146, NG2, and platelet-derived growth factor receptor ␤, but not the smooth muscle cell marker ␣-smooth muscle actin. Coimplantation of hPSC-derived endothelial cells with pericytes resulted in functional and rapid anastomosis to the murine vasculature. Administration of pericytes into immunodeficient mice with limb ischemia promoted significant vascular and muscle regeneration. At day 21 after transplantation, recruited hPSC pericytes were found incorporated into recovered muscle and vasculature. Conclusions-Derivation of vasculogenic and multipotent pericytes from hPSC can be used for the development of vasculogenic models using multiple vasculogenic cell types for basic research and drug screening and can contribute to angiogenic regenerative medicine. (Circulation. 2012;125:87-99.)Key Words: pluripotent stem cells Ⅲ ischemia Ⅲ mesenchymal stem cells Ⅲ pericytes Ⅲ vasculature I n adult tissues, the majority of blood vessels are composed of 3 layers including a luminal inner monolayer of endothelial cells (EC), tunica intimae, an intermediate muscular layer, tunica media, of smooth muscle cells (SMC) and an outer layer of fibroblast-like adventitial cells and connective tissue components, tunica adventitia. Microvessels, including capillaries, precapillary arterioles, postcapillary venues, and collecting venules are composed of internal endothelial layer surrounded by outer coverage of pericytes (also known as Rouget cells or mural cells). 1 Both perivascular SMC and pericytes have been shown to function as critical regulators of vascular development, stabilization, maturation, and remodeling mediated by transforming growth factor ␤ (TGF-␤), platelet-derived growth factor B, or angiopoietin-1. 2,3 Although related in function and anatomic localization, pericytes can be distinguished from SMC on the basis of their characteristic morphology and specific cell marker expression: Whereas SMC form a separate layer of the tunica media in blood vessels, pericytes are physically embedded within the endothelial basement membrane to promote mutual communication with the underlying endothelium. 4 In addition, SMC and the majority of pericytes in multiple human and murine tissue types express ␣-smooth muscle actin (␣-SMA), which is involved in regulatio...

Hematopoietic MicroRNA-126 Protects against Renal Ischemia/Reperfusion Injury by Promoting Vascular Integrity

“…Support for this notion is emerging in the literature, where the role of BM-derived pericytes is increasingly discussed. 59,60 Taken together, the current findings demonstrate that enhancing the vasculogenic potential by overexpression of miR-126 in the hematopoietic compartment protects the kidney from IRI, further confirming the critical role of EC integrity in the progression of kidney disease. Therefore, strategies aimed at improvement of the endogenous vasculogenic potential such as described in this study not only would enhance the mobilization of the vasculogenic progenitors to the injured kidney but also may correct EPC dysfunction in renal disease.…”

Pericytes, mesenchymal stem cells and their contributions to tissue repair

“…Furthermore, Matrigel implantation of human endothelial and pericyte co-culture results in neo-formation of the microvasculature, with perfusion and anastomoses with murine blood vessels (Dar et al, 2012). In this study, pericytes were derived from differentiated human induced pluripotent stem cells (hiPSC) from embryoid bodies, and defined, somewhat controversially as compared to traditional methods, as CD105+/CD31-cells (Dar et al, 2012).…”

“…Cultured, osteogenically-stimulated hiPSC-pericytes have also demonstrated mineral deposition and the formation of bone-like structures when implanted in vivo (Dar et al, 2012). Perivascular cells isolated from the SVF have been shown to generate greater ectopic bone formation in a humanmouse xenograft using biological scaffolds as compared to unsorted SVF, suggesting that these are the crucial SVF cells involved in osteogenesis (James et al, 2012a;James et al, 2012c).…”

“…Induced pericytes are also capable of myogenesis; in the presence of TGF-β and PDGF-BB, hiPSC-derived pericytes cocultured with hiPSC-derived ECs are capable of differentiation into contractile smooth muscle cells in vitro, as demonstrated by expression of the contractile markers SM22 and CNN1 (Orlova et al, 2014a). Moreover, CD105+ pericytes isolated from hiPSC-derived embryoid bodies have been found to aid in ischemic muscle recovery and display direct myogenic differentiation, as indicated by desmin expression, in addition to providing vascular support by integrating with the host vasculature (Dar et al, 2012).…”

Different angiogenic potentials of mesenchymal stem cells derived from umbilical artery, umbilical vein and wharton’s jelly

“…Recently, osteogenic and adipogenic progenitors have also been shown to originate from perivascular niches in vivo and purified pericytes [14][15][16]. In addition, transplantation of purified pericytes can support vasculature and repair damaged tissue [17,18]. These results indicate the therapeutic capacity of perivascular stem cells in postinjury angiogenesis/vasculogenesis.…”