In vitro model of human mammary gland microbial colonization (MAGIC) demonstrates distinctive cytokine response to imbalanced human milk microbiota

Treven, Primož (author), Paveljšek, Diana (author), Kostanjšek, Rok (author), Golob, Majda (author), Bogovič Matijašić, Bojana (author), Mohar Lorbeg, Petra (author)

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.

• Language: English
• Keywords: human milk microbiota, bacterial adherence, cytokine response
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty; VF - Veterinary Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2024
• Number of pages: Str. 1-19
• Numbering: Vol. 12, iss. 3
• PID: 20.500.12556/RUL-166014
• UDC: 636.09:579
• ISSN on article: 2165-0497
• DOI: 10.1128/spectrum.02369-23
• COBISS.SI-ID: 184797955
• Publication date in RUL: 17.12.2024

Record is a part of a journal

• Title: Microbiology spectrum
• Shortened title: Microbiol. spectr.
• Publisher: ASM Press
• ISSN: 2165-0497
• COBISS.SI-ID: 4526968

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: mikrobiologija, mlečna žleza, in vitro model, humano mleko, mikrobiota, citokinski odziv

Projects

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P4-0097
• Name: Prehrana in mikrobna ekologija prebavil

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: Z3-8198
• Name: Karakterizacija in vitro modela za študij mikrobiote mlečne žleze (akronim - MAGIC)