Abstract: In order to produce virus free potato tuber seeds in vitro culture was established using apical meristem of four widely cultivated varieties of Bangladesh namely Diamant, Cardinal, Multa and Lalpakri. Meristem of these varieties of potato were cultured for shoot proliferation and root induction in MS and MS0 medium supplemented with different types and concentration of phytohormones. Among the different growth regulator formulations, 0.5 mg l-1 GA3+0.04 mg l-1 KIN was found to be the best medium for the primary establishment of meristem. The primary established meristems were subcultured on to MSo medium and MS medium containing BA and IBA singly or in combinations. Considering all the treatments singly use of IBA (0.5 mg l-1) is recommended for proper shoot and root development from primary meristem. After DAS-ELISSA test the virus free in vitro grown potato plantlets were using for massive micro propagation. GA3 and KIN singly or in combinations are used for shoot and root multiplication, among all these combinations 2.0 mg l-1 GA3 was found to be the best media for shoot induction for the studied varities. For high frequency of root formation, combination of GA3 (0.1 mg l-1)+KIN (0.1 mg l-1) was most effective. Rooted plantlet were gradually acclimatized and successfully established in the field. Visual evolution of the morphological trials of the tissue culture derived plants showed all plants were found normal and free from virus diseases. Substantial yield increase was observed in meristem derived plants over their source plants.