Alaska, Safran and Spunta are the most important varieties of potatoes used by Tunisian farmers. This study was carried out in four steps. First, study on regeneration of tissue culture protocol was studied using buds as an explant for initiation of culture in MS media supplemented with four different concentrations of an auxin: indole butyric acid (IBA). Growth proliferation showed that optimum regeneration rate was obtained with 0, 5 mg/l of IBA. The regenerated plants were cultured using nodal cuttings as explants for further multiplication. In vitro tuberization involving a combination benzyladenine (BA) and paclobutrazol (PBZ) gave good tuberization rate. Yet, liquid media containing 5 mg/l of BA was optimal to produce microtubers for all cultivars. Microtubers were transplanted in the soil, cultured in glasshouse to produce minitubers, they produces,-7, 2 healthy minitubers/plant. Microtubers cultured in medium containing sucrose (80 g/l) gave best number of minitubers/plant.