In recent years, there has been a growing interest in biostimulants as an alternative to chemicals for safe and sustainable agriculture. The aim of this study was to analyze the effect of biostimulators Regoplant and Charkor (Agrobiotech, Ukraine) on growth and development of micropropagated pear plants (Pyrus communis L. ‘Old Home’ x ‘Farmingdale’) at acclimatization stage. In vitro propagated and rooted plantlets from pear rootstock OHF 333 were acclimatized in a floating system with 100 μl L-1 Regoplant or Charkor. Plantlets with no additional treatments served as control. Data on growth parameters, chlorophyll a fluorescence (ОJIP test) and antioxidant activity were collected 45 days after transplanting to ex vitro conditions. Enrichment of the nutrient solution with biostimulator Charkor (100 μl L-1) in floating system led to the highest survival rate (82.5%) of pear plants, the greatest stem length, number, fresh and dry mass of leaves. Combining innovative approaches such as a floating system and biostimulators would significantly improve the acclimatization and the overall process of micropropagation of fruit plants.