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

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

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.

• Jezik: Angleški jezik
• Ključne besede: human milk microbiota, bacterial adherence, cytokine response
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta ; VF - Veterinarska fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2024
• Št. strani: Str. 1-19
• Številčenje: Vol. 12, iss. 3
• PID: 20.500.12556/RUL-166014
• UDK: 636.09:579
• ISSN pri članku: 2165-0497
• DOI: 10.1128/spectrum.02369-23
• COBISS.SI-ID: 184797955
• Datum objave v RUL: 17.12.2024

Gradivo je del revije

• Naslov: Microbiology spectrum
• Skrajšan naslov: Microbiol. spectr.
• Založnik: ASM Press
• ISSN: 2165-0497
• COBISS.SI-ID: 4526968

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: mikrobiologija, mlečna žleza, in vitro model, humano mleko, mikrobiota, citokinski odziv

Projekti

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P4-0097
• Naslov: Prehrana in mikrobna ekologija prebavil

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: Z3-8198
• Naslov: Karakterizacija in vitro modela za študij mikrobiote mlečne žleze (akronim - MAGIC)