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The aim of this study was to investigate the probiotic properties of 174 Lactobacillus strains isolated from Mongolian dairy products, and particularly their impact on intestinal calcium uptake and absorption. All isolates underwent a first screening based on cell surface hydrophobicity, acid tolerance, tolerance to gastro-intestinal digestion, autoaggregation, adhesion and cytotoxicity against intestinal cells. Six Lactobacillus strains from different species (L. casei, L. kefiranofaciens, L. plantarum, L. fermentum, L. helveticus and L. delbrueckii) were selected, and their impact on intestinal calcium uptake and transport was investigated using Caco-2. Five strains were able to improve total calcium transport after 24 h contact with a differentiated Caco-2 cell monolayer. Concomitantly the L. plantarum strain was able to increase cellular calcium uptake in Caco-2 cells by 10% in comparison to control conditions. To determine which pathway(s) of calcium absorption was modulated by the strains, a qPCR-based study on 4 genes related to calcium/vitamin D metabolism or tight junction integrity was conducted on mucus-secreting intestinal HT-29 MTX cells. The L. plantarum strain modulates the transcellular pathway by regulating the expression of vitamin D receptor (1.79 fold of control) and calcium transporter (4.77 fold of control) while the L. delbrueckii strain acts on the paracellular pathway by modulating claudin-2 expression (2.83 fold of control). This work highlights the impact of Lactobacillus probiotic strains on intestinal calcium absorption and for the first time give some evidence about the cellular pathways involved., Competing Interests: Declaration of Competing Interest CR, MF and MV are affiliated with VF Bioscience SAS, France, a company commercializing lactic acid bacteria strains for use as probiotics or dairy starters., (Copyright © 2020 Elsevier Ltd. All rights reserved.)