Linboko zelula amen eta melanozitoen ko-kultiboaren optimizazioa: hazkuntza medioek melanozito epidermikoen hazkundean eta funtzionaltasunean duten eragina

Abstract

Limbal epithelial stem cells (LESC), which are located in the sclerocorneal limbus (the transition zone between cornea and sclera), precisely in a specialized environment called the limbal niche, are constantly renewing the corneal epithelium. The limbal niche also contains melanocytes, which play a key role in maintaining the state of dedifferentiation and the functionality of said stem cells. When LESCs become dysfunctional, the cornea undergoes conjunctivalization, resulting in decreased transparency and loss of vision. Although autologous LESC transplantation is the most effective treatment, the scarcity of LESCs requires their ex vivo expansion, highlighting the need to seek optimal conditions for this. 

Co-seeding with melanocytes has been recently investigated as a promising approach to maintaining the survival, stemness and functionality of LESCs by mimicking their natural niche. Using as the main axis the investigation of morphology, growth, viability, migration, melanin synthesis and expression of melanocyte markers, we analyzed melanocytes in different culture media. The findings indicated that the commonly used media for LESCs is not adequate to culture melanocytes. In contrast, the modified experimental media enhanced viability and migratory capacity while maintaining the typical melanocyte morphology.

Cholera toxin was replaced with isoproterenol to develop growth media free of xenogenic compounds - chemical substances derived from other species- in order to avoid compounds that may pose risks to cell therapy. Furthermore, studies have shown the importance of specific growth factors and signal molecules such as α-MSH and endothelin-1 for melanocyte survival. Optimization of LESC and melanocyte growth conditions is essential to improve LESC expansion and transplantation therapies.