INVESTIGADORES
CERUTI Julieta Maria
congresos y reuniones científicas
Título:
DEVELOPMENT OF TWO IN VITRO ANGIOGENIC MODELS TO EVALUATE THE PRO-ANGIOGENIC CAPACITY OF DERMAL PAPILLA SPHEROIDS
Autor/es:
OPPENHEIMER, FLORENCIA MAIA; CERUTI, JM; LEIRÓS, GJ; BALAÑÁ, ME
Lugar:
Mar del Plata
Reunión:
Congreso; LXV REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA (SAIC); 2020
Institución organizadora:
SAIC
Resumen:
Tissue-engineered skin represents a useful treatment of deep skininjuries. The neovascularization, essential to supply oxygen and nu-trients to the cells in grafted constructs, remains a major challenge.We have shown that the presence of dermal papilla cells (DPCs) inthese constructs favors the vascularization process, resulting in abetter wound healing and graft take. We also have seen that 3D cul-ture increases the expression of angiogenic factors as VEGF, angio-genin and FGF. Our objective was to fit two models to evaluate theDPC?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. HUVECwere then seeded on the insert and placed into wells. After 8 hours,we observed 12% more migrated cells in the coculture with DPCsthan in the fibroblast and control.For tubulogenic assay, DPC conditioned medium was concentrated10 times and diluted by half in fresh medium. HUVEC were resus-pended in the mixed medium and seeded on Matrigel. We observedan increase in the total tubule?s length and in the number of seg-ments and joints, compared to control. The higher differences wereobtained at 5 hours.To compare the inductive molecules secretion between 2D and 3Dcultures, we looked for the condition in which both systems con-tained 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 same period.Progress was made in the development of two angiogenic modelsthat would allow us to study the influence of 3D culture of DPCson the capacity of endothelial cells to migrate and to form tubulesin vitro. In this way, its use in skin substitutes could favor the vas-cularization of the grafts, favoring the closure of the wound and thegraft take.