Amygdalin has been one of the most popular “alternative cancer cures” in many European and South American countries. Its anticancer, anti-inflammatory activity and other medicinal benefits have been known for many years. The objective of this in vitro study was to examine the potential impact of amygdalin on the cell viability and production of steroid hormone testosterone by porcine ovarian granulosa cells. Granulosa cells were isolated from porcine ovaries and subsequently cultured without (control) or with amygdalin at various doses (1; 10; 100; 1000 and 10 000 μg/mL) for 24 h. The cell viability was determined by alamarBlueTM reagent and release of testosterone was assayed by ELISA. Obtained results showed a significant (P<0.05) increase of testosterone secretion only at the highest dose of amygdalin (10 000 μg/mL). Other experimental doses of amygdalin did not affect the testosterone production. Moreover, amygdalin treatment strongly enhanced the viability of ovarian granulosa cells. The viability was significantly (P<0.05) stimulated after amygdalin treatment at all used doses, except the highest concentration (10 000 μg/mL). To conclude, application of amygdalin to culture media positively affected cell viability, but not highest dose (10 000 µg/mL), and stimulated testosterone release by porcine ovarian cell. Present results could help to reveal the potential impact of amygdalin on cellular growth, as well as its mechanism of action in processes of ovarian steroidogenesis.