The cytological status of callus and regenerants ofPodophyllum hexandrumcultured in vitrofor over a period of 3 years is reported. Calli maintained both on lower (0.5 μM) and higher (10 μM) 2,4-D, remained cytologically stable up to 18 months. With the long term (3 years) culture on higher 2,4-D both numerical and structural changes in the chromosomes were observed. The calli maintained for 3 years on lower 2,4-D on the other hand retained both chromosome stability and regeneration potentiality. The regenerants in general showed normal mitotic complements. One of the regenerants from higher 2,4-D medium, however, showed structural chromosome changes.