In vitro analysis of the interactions between preadipocytes and endothelial cells in a 3D fibrin matrix.
The volume-persistent survival of transplanted adipose tissue in vivo relies on early vascularization, due to an otherwise early induction of apoptosis of the centrally located cells. Thus, one way to enable the early formation of a capillary network resulting in a sufficient perfusion of the transplanted construct might be the co-transplantation of autologous preadipocytes with endothelial cells. To investigate preadipocyte-endothelial cell interaction, three-dimensional proliferation- and angiogenesis assays were performed in vitro. Proliferation rates of co-cultured endothelial cells and preadipocytes suspended in a fibrin matrix were elucidated by Alamarblue assays. The spheroid angiogenesis model was applied for analyzing the effects of vascular endothelial cell growth factor (VEGF) and basic fibroblast growth factor (bFGF) (produced by preadipocytes) as well as the impact of cell-cell interaction between preadipocytes and endothelial cells and fibrin matrix on endothelial cell migration. Preadipocytes proliferated in fibrin glue, whereas endothelial cells underwent apoptosis. By co-culturing, both cell types demonstrated an increased proliferation rate. Preadipocytes provoked migration of endothelial cells. Blocking bFGF and/or VEGF led to a significant decrease of migration. Changes in fibrin structure were followed by migration of single cells instead of sprouting. An appropriate fibrin matrix as well as already differentiated endothelial cells are necessary for preadipocytes to develop their angiogenic activity via bFGF and VEGF.
Borges
Jörg
J
Department of Plastic and Hand Surgery, University of Freiburg Medical Center, Freiburg, Germany. borges@ch11.ukl.uni-freiburg.de
Müller
Matthias C
MC
Momeni
Arash
A
Stark
G Björn
GB
Torio-Padron
Nestor
N
D017667:Adipocytes / Q000166:cytology*; D000273:Adipose Tissue / Q000098:blood supply / Q000166:cytology / Q000637:transplantation; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000166:cytology*; D015718:Fibrin Tissue Adhesive; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D006801:Humans; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010982:Platelet-Derived Growth Factor; D023822:Tissue Engineering / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D015718:Fibrin Tissue Adhesive; D010982:Platelet-Derived Growth Factor; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2