CHARACTERIZATION OF PRIMARY CULTURE OF BOVINE CORPUS LUTEUM (CPCLB): AN EXPERIMENTAL MODEL FOR THE STUDY OF APOPTOSIS AND DIRECT EFFECTS OF GONADOTROPIN-RELEASING HORMONE (GnRH) ANALOGUES.

GERMANO, P.; MOHAMAD, N.; SOÑEZ, MC.; LOMBARDO, DM.

Ciudad Autónoma de Buenos Aires

Jornada; IV Jornadas Internacionales del Instituto de Investigación y Tecnología en Reproducción Animal; 2014

Instituto de Investigación y Tecnología en Reproducción Animal

The CPCLB is an experimental model representative of bovine corpus luteum behavior without infl uence of the hypothalamic-pituitarygonadal axis. Th e objectives of this study were to characterize CPCLB regarding its heterogeneous cellular composition and to evaluate its sensitivity to apoptosis inducing factors for studying the modulation by GnRH analogues. Primary cultures of cells obtained from bovine corpus luteum of slaughtered animals were performed and cell growth curves were obtained. Staining with haematoxylin/eosin and oil red O were made to determine morphological and histochemical features. Th e expression of a)3 hydroxysteroid dehydrogenase (3 HSD), marker of steroidogenic cells, b) Von Willebrand Factor (VWF), marker of endothelial cells, c) active caspase 3 and d) GnRH-I receptor were immunocytochemicaly evaluated. Prevailing cell populations were quantifi ed by fl ow cytometry analysis with 3 HSD and VWF and the response to apoptosis induction by serum deprivation and H2O2 was assessed by TUNEL. Steroidogenic cells were the main cell type present in CPCLB (65-85%) and showed the presence of lipid vacuoles and expression of 3 HSD and GnRH-I receptor. Th e culture also presented smaller endothelial cells (25-35%), positively marked for VWF. Images of cells in interphase, mitosis and apoptosis were observed. Rate of apoptosis increased in response to apoptotic inducers in respect of control, indicating sensitivity of CPCLB to apoptosis. Th e positive expression of GnRH-I receptor, would permit evaluation of the direct eff ect of GnRH analogues on luteal cells via this receptor.