Conditions for protoplast isolation and culture in the dioecious species Melandrium album (Silene alba) are described and differences between female and male plants subjected to in vitro culture identified. Plant regeneration was achieved in the case of female plants only. Selection for high regeneration capacity lines was initiated aiming at creating responsive experimental material for further studies. The regeneration behaviour in this dioecious plant is discussed in the context of hormonal models on sex determination.