PDGF, TGF-β, and Heterotypic Cell–Cell Interactions Mediate Endothelial Cell–induced Recruitment of 10T1/2 Cells and Their Differentiation to a Smooth Muscle Fate

Abstract

We aimed to determine if and how endothelial cells (EC) recruit precursors of smooth muscle cells and pericytes and induce their differentiation during vessel formation. Multipotent embryonic 10T1/2 cells were used as presumptive mural cell precursors. In an under-agarose coculture, EC induced migration of 10T1/2 cells via platelet-derived growth factor BB. 10T1/2 cells in coculture with EC changed from polygonal to spindle-shaped, reminiscent of smooth muscle cells in culture. Immunohistochemical and Western blot analyses were used to examine the expression of smooth muscle (SM)-specific markers in 10T1/2 cells cultured in the absence and presence of EC. SM-myosin, SM22α, and calponin proteins were undetectable in 10T1/2 cells cultured alone; however, expression of all three SM-specific proteins was significantly induced in 10T1/2 cells cocultured with EC. Treatment of 10T1/2 cells with TGF-β induced phenotypic changes and changes in SM markers similar to those seen in the cocultures. Neutralization of TGF-β in the cocultures blocked expression of the SM markers and the shape change. To assess the ability of 10T1/2 cells to contribute to the developing vessel wall in vivo, prelabeled 10T1/2 cells were grown in a collagen matrix and implanted subcutaneously into mice. The fluorescently marked cells became incorporated into the medial layer of developing vessels where they expressed SM markers. These in vitro and in vivo observations shed light on the cell–cell interactions that occur during vessel development, as well as in pathologies in which developmental processes are recapitulated.

Histone Deacetylase 7‐Derived Peptides Play a Vital Role in Vascular Repair and Regeneration

“…In the meantime, the recruitment of SMCs is vital to stabilize the newly formed endothelium 32, 39, 40, 41, 42. In this study, we have demonstrated that the inclusion of 7A, especially the phosphorylated version 7Ap, significantly increased endothelialization.…”

Msx2 promotes cardiovascular calcification by activating paracrine Wnt signals

“…We previously demonstrated that the C3H10T1/2 cell system represents a highly useful cell culture model for studying effects of Msx2 on osteogenic, adipogenic, and smooth muscle differentiation (10). This mesenchymal progenitor shares features of vascular myofibroblasts and can readily adopt the mural VSMC fate when transplanted in vivo (30). Therefore, we assessed whether the conditioned media (CM) from Msx2-transduced 10T1/2 cells could alter osteogenic and/or adipogenic differentiation.…”

Endothelialization of Acellular Porcine ECM with Chemical Modification

“…Mural cells, including vascular smooth muscle cells and pericytes, together with endothelial cells, possess the potential of recruiting cells from the surrounding tissue in vivo [12]. MSCs can be differentiated into both endothelial and mural cells in vitro and in vivo [13]. Our preliminary studies demonstrated that endothelial cells seeded into the vasculature of the ECM were only shown to survive and reach a stabilized phase over time without Endothelialization of Acellular Porcine ECM with Chemical Modification proliferation, which is necessary for the formation of functional blood vessels.…”