In this study, we examined the effect of: coconut water (CW), plant growth regulators NAA ? a-naphthaleneacetic acid, IBA ? b-indolilobutyric acid as well as BAP ? 6-benzyl aminopurine and steroidal hormones (progesterone, estriol, estrone, 17-b-estradiol, androsterone, androstendione) on callus of Polygonatum verticillatum (L.) All. induction. The best callus formation was obtained from the hypocotyl part of plants cultivated from the seeds after their stratification (two weeks in 4C) on the Murashige and Skoog (MS) medium, which contained: 100 mg/dm3 of myoinositol, 30 g/dm3 of sucrose, 2 mg/dm3 2,4-D and 1 mM/dm3 17-b-estradiol. Additionally, the efforts to obtain the callus from underground parts of P. verticillatum were made. It was done on MS medium containing: 1 mg/dm3 2,4-D, 1 mM/dm3 17-b-estradiol and 2 mg/dm3 6 ? BAP. The callus grown in in vitro conditions was used for the fitochemical analysis to determine the presence of diosgenin.