PhD Scientific Days 2025

Theoretical and Translational Medicine II.

Development of an In Vitro Model for Assessing Intestinal Epithelial Permeability

Előadó neve

Vágó-Szincsák Sára, MSc

Neptun code

H1ZP61

Előadó munkahelye

Institute of Translational Medicine, Semmelweis University

Előadó telefonszáma

+36203335216

Előadó e-mail címe

vago.sara@phd.semmelweis.hu

Az előadás címe

Development of an In Vitro Model for Assessing Intestinal Epithelial Permeability

Szerző(k) neve és munkahelye

Sara Vago-Szincsak¹, Csenge Somodi¹, Gabor Szegvari¹, Dora David², Anna Földes³, Zoltan Lohinai^1,4

1: Institute of Translational Medicine, Semmelweis University
2: Department of Anatomy, Histology and Embryology, Semmelweis University
3: Department of Oral Biology, Semmelweis University
4: Pulmonology Hospital Törökbálint

Bemutatás módja

Szóbeli

Szekció

Theoretical and Translational Medicine II.

Language of the presentation

English

Preferred session

Theoretical and Translational Medicine

Összefoglaló szövege

Introduction: Urine samples provide a non-invasive method for studying potential circulating biomarkers originating from the host organism. Specific proteins can cross the intestinal barrier and may be derived from the gut microbiota. This approach supports the identification of biomarkers for early diagnosis and personalized treatment strategies. Establishing a reliable in vitro model is crucial for simulating the intestinal environment, enabling researchers to investigate how various substances interact with the epithelial barrier and affect its permeability. Aims: We aimed to create an epithelial monolayer to assess the gut barrier integrity and study the effect of identified urine EV proteins on tight junctions. Method: Human epithelial T84 and LS174T cell lines were grown in a Transwell setup on a membrane with a pore size of 0.4 μm. The integrity of the cellular layer was evaluated using transepithelial electrical resistance (TEER) measurements. Immunofluorescence staining was employed to investigate the distribution of MUC2 and ZO-1 proteins. Results: Co-cultures containing increasing proportions of LS174T cells demonstrated decreased maximum TEER values, indicating reduced barrier integrity. A higher number of seeded cells in T84 cultures on non-coated inserts led to increased resistance. In LPS-treated cultures, TEER decreased in a dose- and time-dependent manner, with 10 ng/mL inducing a 20% drop after 180 minutes. Immunofluorescent imaging revealed distinct mucin layer formation in co-culture conditions, whereas tight junction structures were less defined than in T84 monocultures. Conclusion: We established a customizable in vitro model for studying intestinal barrier properties. Co-culture systems with goblet-like cells produce physiologically relevant mucin layers but show decreased electrical resistance, limiting TEER as a sole endpoint. FITC-dextran permeability assays and further cytokine profiling are planned to complement functional barrier integrity assessment. Funding: This study was supported by the Semmelweis 250+ Excellence PhD Scholarship and the Hungarian National Research, Development and Innovation Office (OTKA-FK #146775; OTKA-PD #142287).

University

Semmelweis University

Supervisor

Zoltan Lohinai MD, PhD

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies after complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem hagyta jóvá

Előadó

7486

Start

12:45

End

13:00