The complex metabolism of probiotic bacteria requires several technological options to guarantee the functionally of probiotic dairy foods during the shelf life. This research aimed to evaluate the effect of the supplementation of increasing amounts of Lactobacillus acidophilus (0, 0.4, or 0.8 g/L of milk) on the physicochemical parameters and sensory acceptance of Minas fresh cheese. In addition, the sensory acceptance of probiotic cheeses was assessed using a consumer test and compared with commercial cheeses (conventional and probiotic). High counts (9.11 to 9.42 log cfu/g) of L. acidophilus were observed throughout the shelf life, which contributed to the maintenance of its probiotic status and resulted in lower pH values and greater production of organic acids. The probiotic cheeses presented lower scores for appearance, aroma, and texture compared with conventional cheeses. Internal preference mapping explained almost 130% of the total variation of the data and showed a large number of consumers concentrated near the conventional cheeses, demonstrating greater preference for these samples. The findings indicated that some negative sensory effects could occur when high level of supplementation with L. acidophilus is used in probiotic cheese processing.941047774786