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

doi:10.1083/jcb.141.3.805

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© The Rockefeller University Press, 0021-9525/1998//805 $5.00
The Journal of Cell Biology, Volume 141, Number 3, , 1998 805-814

Articles

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

Karen K. Hirschi, Stephanie A. Rohovsky, and Patricia A. D'Amore

Harvard Medical School and Children's Hospital, Boston, Massachusetts 02115

We aimed to determine if and how endothelial cells (EC) recruitprecursors of smooth muscle cells and pericytes and inducetheir differentiation during vessel formation. Multipotentembryonic 10T1/2 cells were used as presumptive mural cellprecursors. In an under-agarose coculture, EC induced migrationof 10T1/2 cells via platelet-derived growth factor BB. 10T1/2cells in coculture with EC changed from polygonal to spindle-shaped,reminiscent of smooth muscle cells in culture. Immunohistochemicaland Western blot analyses were used to examine the expressionof smooth muscle (SM)-specific markers in 10T1/2 cells culturedin the absence and presence of EC. SM-myosin, SM22 ${alpha}$ , and calponinproteins were undetectable in 10T1/2 cells cultured alone;however, expression of all three SM-specific proteins was significantlyinduced in 10T1/2 cells cocultured with EC. Treatment of 10T1/2 cells with TGF-β induced phenotypic changes and changesin SM markers similar to those seen in the cocultures. Neutralizationof TGF-β in the cocultures blocked expression of the SMmarkers and the shape change. To assess the ability of 10T1/2cells to contribute to the developing vessel wall in vivo, prelabeled 10T1/2 cells were grown in a collagen matrix and implantedsubcutaneously into mice. The fluorescently marked cells becameincorporated into the medial layer of developing vessels wherethey expressed SM markers. These in vitro and in vivo observationsshed light on the cell–cell interactions that occur duringvessel development, as well as in pathologies in which developmentalprocesses are recapitulated.

Abbreviations used in this paper: Ac-LDL, acetylated low density lipoprotein; BAE, bovine aortic EC; CS, calf serum; EC, endothelial cells; SM, smooth muscle; SMC, smooth muscle cells.

Address all correspondence to Patricia A. D'Amore, Laboratoryfor Surgical Research, Children's Hospital, Enders 1061, 300Longwood Ave., Boston, MA 02115. Tel.: 617-355-8377. Fax: 617-355-7043.E-mail: damore_p{at}a1.tch.harvard.edu

The current address of Karen K. Hirschi is Children's NutritionResearch Center, Baylor College of Medicine, 1100 Bates Street,Houston, TX 77030. The current address of Stephanie A. Rohovskyis Department of Surgery, Beth Israel-Deaconess Hospital, Boston,Massachusetts 02115.

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Chen, S., Kulik, M., Lechleider, R. J. (2003). Smad proteins regulate transcriptional induction of the SM22{alpha} gene by TGF-{beta}. Nucleic Acids Res 31: 1302-1310 [Abstract] [Full Text]
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