Long term culture and characterization of European sea bass (Dicentrarchus labrax) ovarían follicular cells

Trabajo presentado en el 8º Congreso de la Asociación Ibérica de Endocrinología Comparada celebrado en Madrid (España) del 5 al 7 septiembre 2011Ovarian follicular cells have a crucial role in reproductive success as they support oocyte growth and maturation. Absence of European sea bass immortalized cell lines led us to develop a primary culture suitable for functional studies. This in vitro model permits analysis at the cellular level and is easy to handle, although cell availability and variability between preparations remain limiting. The primary culture for sea bass follicular cells set up in our laboratory was studied in terms of steroid production, characteristic gene expression and response to different stimuli. Simultaneously, the cell dispersion protocol was modified to perform three independent immortalization trials using classical trypsin dissociation solution prior to subculture. Modifications of culture conditions (temperature, serum type and hormonal treatments) were tested to improve proliferation and survival rates. Even though extreme variability was observed, to date, follicular cells have survived 200 days and have been passed up to 10 times. Follicular primary cell culture is actually used as an in vitro model in our laboratory to carry out studies on promoter regulation and follicular cell physiology, to better understand the process of follicular growth and differentiation