title: Perivascular-like cells contribute to the stability of the vascular network of osteogenic tissue formed from cell sheet-based constructs.
abstract: In recent years several studies have been supporting the existence of a close relationship in terms of function and progeny between Mesenchymal Stem Cells (MSCs) and Pericytes. This concept has opened new perspectives for the application of MSCs in Tissue Engineering (TE), with special interest for the pre-vascularization of cell dense constructs. In this work, cell sheet technology was used to create a scaffold-free construct composed of osteogenic, endothelial and perivascular-like (CD146(+)) cells for improved in vivo vessel formation, maturation and stability. The CD146 pericyte-associated phenotype was induced from human bone marrow mesenchymal stem cells (hBMSCs) by the supplementation of standard culture medium with TGF-β1. Co-cultured cell sheets were obtained by culturing perivascular-like (CD146(+)) cells and human umbilical vein endothelial cells (HUVECs) on an hBMSCs monolayer maintained in osteogenic medium for 7 days. The perivascular-like (CD146(+)) cells and the HUVECs migrated and organized over the collagen-rich osteogenic cell sheet, suggesting the existence of cross-talk involving the co-cultured cell types. Furthermore the presence of that particular ECM produced by the osteoblastic cells was shown to be the key regulator for the singular observed organization. The osteogenic and angiogenic character of the proposed constructs was assessed in vivo. Immunohistochemistry analysis of the explants revealed the integration of HUVECs with the host vasculature as well as the osteogenic potential of the created construct, by the expression of osteocalcin. Additionally, the analysis of the diameter of human CD146 positive blood vessels showed a higher mean vessel diameter for the co-cultured cell sheet condition, reinforcing the advantage of the proposed model regarding blood vessels maturation and stability and for the in vitro pre-vascularization of TE constructs.
authors: 3B's Research Group-Biomaterials, Biodegradables and Biomimetics, University of Minho, Guimarães, Portugal.
authors: Luís F
authors: LF
authors: Mendes
authors: Rogério P
authors: RP
authors: Pirraco
authors: Wojciech
authors: W
authors: Szymczyk
authors: Ana M
authors: AM
authors: Frias
authors: Tírcia C
authors: TC
authors: Santos
authors: Rui L
authors: RL
authors: Reis
authors: Alexandra P
authors: AP
authors: Marques
mesh_terms: D051929:CD146 Antigen / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000502:physiology*; D020286:Pericytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
chemical_list: D051929:CD146 Antigen; D053773:Transforming Growth Factor beta1