The research aimed to evaluate the effect of planting material and B1 vitamins during acclimatization on potato growth. The research was conducted at greenhouse PT. Adhiguna Jaya Laboratory, Central Java - Indonesia. The study used a completely randomized design - factorial 2 x 3 with four replications. The first factor was planting material: cuttings shoot plantlet (H1) and whole plantlet (H2). The second factor was an application of B1 vitamin in growing media : 0 ml. L−1 (M1), 1 ml. L−1 (M2), and 2 ml. L−1 (M3). The results showed that the whole plantlet had a significant effect on plant height, the number of leaves, shoots, and root length than the cuttings shoot planlet. Application of B1 vitamin 1 ml. L−1 at growing media showed the highest effect on roots length. Combination of whole plantlet and concentration B1 vitamin at 1 ml. L−1 - 2 ml. L−1 were increased growth the highest of 95.24% - 100% number of leaves, 2.50% - 3.00% shoots, and 15.82% - 19.90% root length. It can be concluded that the whole plantlet and application of 1 ml. L−1 - 2 ml. L−1 B1 vitamin in growing media during acclimatization increased the growth of potatoes.