DEVELOPMENT OF TWO IN VITRO ANGIOGENIC MODELS TO EVALUATE THE PRO-ANGIOGENIC CAPACITY OF DERMAL PAPILLA SPHEROIDS

OPPENHEIMER, FLORENCIA MAIA; CERUTI, JULIETA MARÍA; LEIROS, GUSTAVO JOSÉ; BALAÑA, MARIA EUGENIA

Congreso; REUNIÓN DE SOCIEDADES DE BIOCIENCIAS 2020; 2020

Tissue-engineered skin represents a useful treatment of deep skin injuries. The neovascularization, essential to supply oxygen and nutrients to the cells in grafted constructs, remains a major challenge.We have shown that the presence of dermal papilla cells (DPCs) in these constructs favors the vascularization process, resulting in a better wound healing and graft take. We also have seen that 3D culture increases the expression of angiogenic factors as VEGF, angiogenin and FGF. Our objective was to fit two models to evaluate the DPC?s culture influence over the ability of endothelial cells (HUVEC) to migrate and form tubules in vitro For the migration assay, isolated DPC or fibroblasts (Fbs) were cultured in a transwell and maintained at 37°C for 72 hours. HUVEC were then seeded on the insert and placed into wells. After 8 hours, we observed 12% more migrated cells in the coculture with DPCs than in the fibroblast and control.For tubulogenic assay, DPC conditioned medium was concentrated 10 times and diluted by half in fresh medium. HUVEC were resuspended in the mixed medium and seeded on Matrigel. We observed an increase in the total tubule?s length and in the number of segments and joints, compared to control. The higher differences were obtained at 5 hours.To compare the inductive molecules secretion between 2D and 3Dcultures, we looked for the condition in which both systems contained approximately the same amount of metabolically active cells.We obtained that 55 spheres of 5,000 cells each with 72 formationhours are metabolically equivalent to 10,000 cells/cm2 monolayerseeded cells, cultured during the sam e period. Progress was made in the development of two angiogenic models that would allow us to study the influence of 3D culture of DPCs on the capacity of endothelial cells to migrate and to form tubulesin vitro. In this way, its use in skin substitutes could favor the vascularization of the grafts, favoring the closure of the wound and the graft take