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Vascularization and engraftment of a human skin substitute using circulating progenitor cell-derived endothelial cells

Benjamin R. Shepherd^*,, David R. Enis^*,, Feiya Wang^,,1, Yajaira Suarez^*,, Jordan S. Pober^*,,,,2 and Jeffrey S. Schechner^,,3

^* Department of Pathology,

Department of Dermatology,

Department of Immunobiology, and

Interdepartmental Program in Vascular Biology and Transplantation, Yale University School of Medicine, New Haven, Connecticut, USA

²Correspondence: Yale University School of Medicine, 295 Congress Ave., Boyer Center for Molecular Medicine Rm. 454, New Haven, CT 06510, USA. E-mail: jordan.pober{at}yale.edu

ABSTRACT

We seeded tissue engineered human skin substitutes with endothelial cells (EC) differentiated in vitro from progenitors from umbilical cord blood (CB-EC) or adult peripheral blood (AB-EC), comparing the results to previous work using cultured human umbilical vein EC (HUVEC) with or without Bcl-2 transduction. Vascularized skin substitutes were prepared by seeding Bcl-2-transduced or nontransduced HUVEC, CB-EC, or AB-EC on the deep surface of decellularized human dermis following keratinocyte coverage of the epidermal surface. These skin substitutes were transplanted onto C.B-17 SCID/beige mice receiving systemic rapamycin or vehicle control and were analyzed 21 d later. CB-EC and Bcl-2-HUVEC formed more human EC-lined vessels than AB-EC or control HUVEC; CB-EC, Bcl-2-HUVEC, and AB-EC but not control HUVEC promoted ingrowth of mouse EC-lined vessels. Bcl-2 transduction increased the number of human and mouse EC-lined vessels in grafts seeded with HUVEC but not with CB-EC or AB-EC. Both CB-EC and AB-EC-induced microvessels became invested by smooth muscle cell-specific alpha-actin-positive mural cells, indicative of maturation. Rapamycin inhibited ingrowth of mouse EC-lined vessels but did not inhibit formation of human EC-lined vessels. We conclude that EC differentiated from circulating progenitors can be utilized to vascularize human skin substitutes even in the setting of compromised host angiogenesis/vasculogenesis.—Shepherd, B. R., Enis, D. R., Wang, F., Suarez, Y., Pober, J. S., Schechner, J. S. Vascularization and engraftment of a human skin substitute using circulating progenitor cell-derived endothelial cells.

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