In order to achieve positive health benefits to the host, metabolically active live probiotic cells need to be administrated in exactly defined numbers of cells, as concentrated formulations, which can be prepared by freeze-drying. Hence, in this work the freeze-drying of the wet biomass of the probiotic strains Enterococcus faecium A7 and Lactobacillus fermentum A8 was performed and moreover an influence of the different lyoprotectors on the bacterial viability during freeze-drying was observed. Several different lyoprotectors were investigated including: glycerol, skim milk, calcium carbonate, sorbitol, inulin, lactose and sucrose. The number of cells before and after freeze-drying was determined by indirect method and the percentage of the cell survival was calculated. The results have shown that the addition of the lyoprotectans immediately before freeze-drying has positively influenced on the percentage of survival of lactic acid bacteria and therefore has protective role. The addition of the skim milks has shown the highest percentages of the cell viability compared to the viability of the cells achieved in the cases when the rest of the tested lyoprotectors were applied. These values were high for the both probiotic strains E. faecium A7 is achieved 98,5%, and for L. fermentum A8 97,3% respectively. Freeze-drying can be successfully applied for the production and storage of probiotic strains as it enables the maintenance of the cell viable count in exactly defined amounts which are necessary in order to exert health benefits to the host, aimed for their application as living drugs.