© 2022 American Dairy Science AssociationThe use of polymer blends as carriers for probiotic cells or using multi-strain probiotic culture mixture in film formulations has a high potential to maintain the stability of probiotics throughout storage. In this study, the survival of Lactobacillus acidophilus, Lactobacillus plantarum, and mixed culture (Lactobacillus spp., Lactococcus spp., and Bifidobacterium spp.) in whey protein isolate (W), carrageenan (C), and W/C blend (W to C on a wt/wt basis at 100 to 0, 75 to 25, 50 to 50, and 0 to 100) films were investigated during 30 d of storage at 4 and 25°C. The water vapor, mechanical, optical, and morphological properties of film samples were also determined. A significant decrease in total lactic acid bacteria counts of all strains (5–6 log cfu/g in reduction) for W and C films was observed during storage at 25°C, whereas blended films had 2 to 3 log cfu/g reduction. The mixed culture-incorporated films had higher cell counts during all storage temperatures. The incorporation of probiotic bacteria significantly influenced the water vapor permeability and color values of films while decreasing tensile strength and elongation at break values. This study reveals that a multi-strain mixed culture presented more chance for survival inside the polymer matrix, especially when carbohydrate- and protein-based polymers were blended.