Aims of this study were: to investigate whether FSH and LH secretion from in-vitro cultured buffalo adenohypophyseal cells can be increased by supplementing culture media with diethyl-stilbestrol and medroxy-progesterone, respectively; to monitor bioactivity of these in-vitro produced gonadotropins and to see if these gonadotropins have any adverse effects on hematology and internal body organs of female rabbits. Pituitary glands collected from 36 adult buffaloes slaughtered at a local abattoir were used. The anterior pituitary cells were cultured in-vitro using medium RPMI-1640 (code R6504-Sigma) enriched with 10% fetal calf serum and GnRH and treated with 0.5 or 1.0 mg/100 ml diethyl-stilbestrol, and 2.5 or 5.0 mg/ml medroxy-progesterone, or left as untreated control. The results showed that FSH and LH concentrations from cultures treated with low or high dose of respective steroids were higher (P<0.05) than those for controls. Treatment of pre-pubertal female rabbits with in-vitro extracted FSH increased serum FSH and LH concentrations, ovarian size and number of developing follicles (GFs) on the ovaries compared to controls (P<0.01). However, rabbits treated with in-vitro produced extract of LH showed increased serum FSH and LH, while there was no effect on ovarian size and number of GFs. Moreover, treatment of rabbits with both gonadotropins had no effects on body weight, hematological variables and internal body organs. In conclusion, diethyl-stilbestrol and medroxy-progesterone enhanced the secretion of FSH and LH, respectively, from cultured pituitary cells. Moreover, in-vitro produced FSH increased ovarian size, serum FSH and LH and stimulated ovarian activity, while in-vitro produced LH neither increased ovarian size nor stimulated ovarian activity.

Key words: Adenohypophyseal cells, Bioactivity, Diethyl-stilbestrol, Gonadotropins, in-vitro culture, Medroxy-progesterone