Caracterização fenotípica e potencial de diferenciação neuronal de células-tronco estromais da córnea humana para aplicações em engenharia tecidual

Abstract

Given the ethical and translational restrictions associated with animal models in ocular research, the use of stem cells emerges as a valuable alternative. These cells offer broad advantages in biotechnological applications, allowing the development of in vitro models that are more representative of human physiology. This study aimed to phenotypically characterize human corneal stromal stem cells (hCSSCs) obtained from corneas discarded for transplantation and to investigate their potential for neural differentiation in vitro. hCSSCs were isolated and subjected to cell expansion, flow cytometry, multipotency assays, and neural differentiation protocols. The cells presented an immunophenotypic profile compatible with mesenchymal lineages (CD73+/CD90+/CD105+) and an absence of hematopoietic markers (CD34-/CD45-). During neural differentiation, a progressive reduction in the expression of the progenitor markers Nestin and Pax6 was observed, associated with an increase in the expression of βIII-tubulin, indicating neuronal maturation. Furthermore, the cells demonstrated the ability to organize into three-dimensional spheroids. The results indicate that discarded corneas constitute a viable and low-cost source for obtaining hCSSCs, highlighting the potential of these cells for applications in vitro modeling of ocular neuropathies and regenerative medicine.