20.500.12556/RUL-166014 In vitro model of human mammary gland microbial colonization (MAGIC) demonstrates distinctive cytokine response to imbalanced human milk microbiota Despite the established concept of the human mammary gland (MG) as a habitat with its own microbiota, the exact mechanism of MG colonization is still elusive and a well-characterized in vitro model would reinforce studies of the MG microbiota development. We aimed to establish and characterize an in vitro cell model for studying MAmmary Gland mIcrobial Colonization (MAGIC) model. We used the immortalized cell line MCF10A, which expresses the strong polarized phenotype similar to MG ductal epithelium when cultured on a permeable support (Transwell). We analyzed the surface properties of the MAGIC model by gene expression analysis of E-cadherin, tight junction proteins, and mucins and by scanning electron microscopy. To demonstrate the applicability of the model, we tested the adhesion capability of the whole human milk (HM) microbial community and the cellular response of the model when challenged directly with raw HM samples. MCF10A on permeable supports differentiated and formed a tight barrier, by upregulation of CLDN8, MUC1, MUC4, and MUC20 genes. The surface of the model was covered with mucins and morphologically diverse with at least two cell types and two types of microvilli. Cells in the MAGIC model withstood the challenge with heat-treated HM samples and responded differently to the imbalanced HM microbiota by distinctive cytokine response. The microbial profile of the bacteria adhered on the MAGIC model reflected the microbiological profile of the input HM samples. The well-studied MAGIC model could be useful for studies of bacterial attachment to the MG and for in vitro studies of biofilm formation and microbiota development. human milk microbiota bacterial adherence cytokine response mikrobiologija mlečna žleza in vitro model humano mleko mikrobiota citokinski odziv true false false Angleški jezik Slovenski jezik Članek v reviji 2024-12-17 15:17:45 2024-12-17 15:17:49 2024-12-18 03:57:10 0000-00-00 00:00:00 2024 0 0 Str. 1-19 iss. 3 Vol. 12 2024 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 636.09:579 2165-0497 10.1128/spectrum.02369-23 184797955 treven-et-al-2024-in-vitro-model-of-human-mammary-gland-microbial-colonization-(magic)-demonstrates-distinctive.pdf treven-et-al-2024-in-vitro-model-of-human-mammary-gland-microbial-colonization-(magic)-demonstrates-distinctive.pdf 1 AC69D3BE4222C7E35B7696C2BC72FACF 2d6901f8d75fcd8a68c6204a4f4a2dcfdf3df3091b9fbabf73f70ac78f1e458c 44586c60-bc82-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=197379 https://journals.asm.org/doi/10.1128/spectrum.02369-23 1 28d3cd32-bc81-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=197378 Biotehniška fakulteta Veterinarska fakulteta 0 0 0