Amygdalin, a natural substance, is a cyanogenic glycoside occurring in the seeds of apricots and bitter almonds. It is a controversial anti-tumor compound that has been used as an alternative cancer drug for many years. Amygdalin is composed of two molecules of glucose, one of benzaldehyde, which induces an analgesic action, and one of hydrocyanic acid, which is an anti-neoplastic compound. This in vitro study was performed to evaluate the possible impact of amygdalin (1, 10, 100, 1000, 10 000 μg/mL) on the secretory activity of granulosa cells (GCs) from porcine cyclic ovaries. The release of progesterone and estradiol-17β by GCs were evaluated by ELISA. In our study, the noticeable changes in estradiol-17β release by ovarian GCs were determined after the amygdalin addition. Amygdalin, at the highest dose (10 000 μg/mL), significantly (P≤0.05) stimulated the release of estradiol-17β by GCs, in comparison to the untreated control cells. On the contrary, no significant (P≥0.05) changes in the progesterone release by GCs caused by amygdalin addition were observed. In conclusion, obtained results showed that the amygdalin application (various doses) to ovarian GCs caused a dose-dependent stimulation of the estradiol-17β release, but not progesterone, and its possible modulatory impact on the steroid production in porcine ovaries.