Vascularization and engraftment of a human skin substitute using circulating progenitor cell-derived endothelial cells.
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
Benjamin R
BR
Department of Pathology, Yale University School of Medicine, 295 Congress Ave., Boyer Center for Molecular Medicine Rm. 454, New Haven, Connecticut 06510, USA.
Enis
David R
DR
Wang
Feiya
F
Suarez
Yajaira
Y
Pober
Jordan S
JS
Schechner
Jeffrey S
JS
D000818:Animals; D042783:Endothelial Cells / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D018919:Neovascularization, Physiologic; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D020123:Sirolimus / Q000494:pharmacology; D017768:Skin, Artificial; D013234:Stem Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous
D019253:Proto-Oncogene Proteins c-bcl-2; D020123:Sirolimus