II Jornadas Doctorales de la Universidad de Murcia BI-CO-03 In vitro culture of gilthead seabream (Sparus aurata) skin cells D. Ceballos-Francisco , H. Cordero, M. Á. Esteban. Fish Innate Immune System Group. Department of Cell Biology and Histology. Faculty of Biology, Campus Regional de Excelencia Internacional “Campus Mare Nostrum”, University of Murcia, 30100 Murcia, Spain (dianacecilia.ceballos@um.es) Teleost skin is a living non keratinised tissue. Its outermost layer, the epidermis, performs an important protective barrier against environmental changes, external aggressors and phathogen [1]. Fish epidermis is composed by several cell types which allow animals to survive in a harmful milleau; perhaps the most important type of these cells is the motile keratocyte. Keratocytes can cover fish skin wound surfaces with a new protective layer of cells within hours after wounding [2,3,4]. In addition to keratocytes, other cell types are present in fish epidermis acting as important contributors to the fish innate immune response [2]. Interest in fish skin has increased in last decade because of its extraordinary mucosal immunity its potent antimicrobial activity and its high tissue regenerative ability [5,6]. Therefore it is neccesary to study the in vitro behavior of skin cells to understand how them can carry out so many important functions. This work used the gilthead seabream (Sparus aurata) as a model due to its significant commercial value on the marine aquaculture. Our aim was to establish and evaluate the epidermal cell culture at different incubation times. Our results showed an active cell migration of high viable cell at the beginning of the explant culture. Those cells maintained their morphological characteristics and viability after 3 days in culture. This is useful to optimize the culture protocol of fish skin cells as a tool for developing future in vitro approaches. References [1] Vieira, F. A., Gregório, S. F., Ferraresso, S., Thorne, M. A., Costa, R., Milan, M., & Power, D. M. (2011). Skin healing and scale regeneration in fed and unfed sea bream, Sparus auratus. BMC genomics, 12(1). [2] Le Guellec, D., Morvan-Dubois, G., & Sire, J. Y. (2004). Skin development in bony fish with particular emphasis on collagen deposition in the dermis of the zebrafish (Danio rerio). International Journal of Developmental Biology, 48, 217-232. [3] Esteban, M.A. (2012). An overview of the immunological defenses in fish skin. ISRN Immunology, 2012 (29). [4] Johnson, R. M., & Brown, E.J. (2000). Cell-mediated immunity in host defense against infectious diseases. Mandell, Douglas and Bennett's principles and practice of infectious diseases. 5th ed. Philadelphia: Churchill Livingstone,2000, 112-46. [5] Rakers, S., Gebert, M., Uppalapati, S., Meyer, W., Maderson, P., Sell, A. F., & Paus, R. (2010). ‘Fish matters’: the relevance of fish skin biology to investigative dermatology. Experimental dermatology, 19(4), 313-324. [6] Nakatani, Y., Kawakami, A., & Kudo, A. (2007). Cellular and molecular processes of regeneration, with special emphasis on fish fins. Development, growth & differentiation, 49(2), 145-154. Acknowledgements H.C. and D.C.F. wish to thank the Spanish Ministry of Economy and Competitiveness for both F.P.I. scholarships. The financial support of the Spanish Ministry of Economy and Competitiveness (grant numbers AGL2011-30381- C03-01 and AGL2014-51839-C5-1-R) and the Fundación Séneca de la Región de Murcia (grant no. 19883/GERM/15, Grupo de Excelencia de la Región de Murcia, Spain) is also acknowledged.