THREE-DIMENSIONAL CULTURE OF DPC CELLS IMPROVES HUVEC ENDOTHELIAL CELLS ANGIOGENIC CAPABILITIES

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

Virtual

Congreso; Reunion Conjunta SAIC-SAI-AAFE -NANOMED; 2021

SAIC

THREE-DIMENSIONAL CULTURE OF DPC CELLS IMPROVES HUVEC ENDOTHELIAL CELLS ANGIOGENIC CAPABILITIESFlorencia Maia Oppenheimer, María Julieta Ceruti, Gustavo José Leirós, María Eugenia BalañáInstituto de Ciencia y Tecnología Dr. César Milstein (CONICET), Buenos Aires, ArgentinaTissue-engineered skin represents a useful strategy for the treatment of deep skin injuries. However, its correct vascularization 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 DPC-spheres culture increase the expression of angiogenic genes as VEGF, angiogenin and FGF. Since angiogenesis in scaffolds is essential for grafts to survive and integrate with existing host tissue, our aim was to compare the neovascularization abilty of monolayer and spheres cultures of DPC by using two adapted angiogenic modelsTo be able to compare the inductive molecules secretion between monolayer and spheres, we looked for the condition in which both systems contained the same amount of metabolically active cells. In our culture conditions, 45 spheres were metabolically equivalent to 104 cells/cm2 monolayer seeded cells. We observed that serum concentration is critical to sphere formation, when generated by the hanging drop technique. As at least 5% was required for DPC to aggregate, the same conditions were used for monolayer culture, in order to compare the angiogenic effects. For the migration assay, conditioned media by monolayer or spheres were transferred to the bottom of a transwell plate and the HUVEC were seeded on the insert and placed into wells. We obtained ~10% more HUVEC cells migrated in the culture medium conditioned by the spheres than in the culture medium conditioned by the monolayer.In conclusion, three-dimensional culture of DPCs improved the migration ability of HUVEC in vitro. In this way, the use of DPC spheres in skin substitutes may favor the vascularization of the grafts, favoring the closure of the wound and the graft take.