A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut

Tselekouni, Paraskevi; Mohseni-Garakani, Mansoureh; Papa, Steve; Kim, Seong Yeon; Avramoglu, Rita Kohen; Wertheimer, Michael R.; Ajji, Abdellah; Lakatos, Peter L.; Rosenzweig, Derek H.

doi:10.3390/gels12010070

Open AccessArticle

A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut

by

Paraskevi Tselekouni

^1,2,

Mansoureh Mohseni-Garakani

¹

,

Steve Papa

²

,

Seong Yeon Kim

¹,

Rita Kohen Avramoglu

²,

Michael R. Wertheimer

³,

Abdellah Ajji

⁴,

Peter L. Lakatos

^2,5,*,†

and

Derek H. Rosenzweig

^1,6,7,*,†

¹

Department of Surgery, McGill University, Montreal, QC H3A 0G4, Canada

²

Department of Medicine, Division of Gastroenterology, McGill University, Montreal, QC H3A 0G4, Canada

³

Department of Engineering Physics, Polytechnique Montreal, Montreal, QC H3T 1J4, Canada

⁴

Department of Chemical Engineering, Polytechnique Montreal, Montreal, QC H3T 1J4, Canada

⁵

Department of Internal Medicine and Oncology, Semmelweis University, 1428 Budapest, Hungary

⁶

Surgical & Interventional Sciences Program, The Research Institute of McGill University Health Centre, Montreal, QC H4A 3J1, Canada

⁷

Department of Orthopedic Surgery, University of Connecticut School of Medicine, Farmington, CT 06032, USA

^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Gels 2026, 12(1), 70; https://doi.org/10.3390/gels12010070

Submission received: 18 December 2025 / Revised: 5 January 2026 / Accepted: 7 January 2026 / Published: 13 January 2026

(This article belongs to the Special Issue Hydrogels for the Regeneration of Joints of the Musculoskeletal System in Orthopedics (2nd Edition))

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Abstract

Inflammatory bowel disease (IBD) arises from chronic dysregulation at the epithelial–stromal interface, creating a need for in vitro systems that better capture these interactions. In this study, we developed a 3D co-culture platform in which HT-29 intestinal epithelial cells and IMR-90 fibroblasts are embedded within an alginate–gelatin hydrogel, alongside a complementary interface model using a plasma-treated electrospun mesh to spatially compartmentalize stromal and epithelial layers. We first assessed metabolic activity, viability, and proliferation across several epithelial-to-fibroblast ratios and identified 1:0.5 as the most supportive of epithelial expansion. The A1G7 hydrogel maintained high viability (>92%) and sustained growth in all mono- and co-cultures. To evaluate inflammatory competence, models were stimulated with lipopolysaccharide (LPS), administered either within the hydrogel or through the culture medium. LPS exposure increased TNF-α and IL-1β secretion in both configurations, with the magnitude of the response depending on the delivery route. Treatment with dexamethasone consistently reduced cytokine levels, confirming the model’s suitability for pharmacological testing. Together, these results demonstrate that the alginate–gelatin system provides a reproducible epithelial–stromal platform with quantifiable inflammatory readouts, offering a practical foundation for mechanistic studies and early-stage screening of anti-inflammatory therapeutics in IBD.

Keywords: hydrogel scaffold; 3D co-culture model; intestinal epithelium; epithelial–stromal interactions; electrospun scaffold; inflammatory bowel disease; LPS stimulation; anti-inflammatory response

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MDPI and ACS Style

Tselekouni, P.; Mohseni-Garakani, M.; Papa, S.; Kim, S.Y.; Avramoglu, R.K.; Wertheimer, M.R.; Ajji, A.; Lakatos, P.L.; Rosenzweig, D.H. A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut. Gels 2026, 12, 70. https://doi.org/10.3390/gels12010070

AMA Style

Tselekouni P, Mohseni-Garakani M, Papa S, Kim SY, Avramoglu RK, Wertheimer MR, Ajji A, Lakatos PL, Rosenzweig DH. A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut. Gels. 2026; 12(1):70. https://doi.org/10.3390/gels12010070

Chicago/Turabian Style

Tselekouni, Paraskevi, Mansoureh Mohseni-Garakani, Steve Papa, Seong Yeon Kim, Rita Kohen Avramoglu, Michael R. Wertheimer, Abdellah Ajji, Peter L. Lakatos, and Derek H. Rosenzweig. 2026. "A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut" Gels 12, no. 1: 70. https://doi.org/10.3390/gels12010070

APA Style

Tselekouni, P., Mohseni-Garakani, M., Papa, S., Kim, S. Y., Avramoglu, R. K., Wertheimer, M. R., Ajji, A., Lakatos, P. L., & Rosenzweig, D. H. (2026). A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut. Gels, 12(1), 70. https://doi.org/10.3390/gels12010070

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A 3D Alginate–Gelatin Co-Culture Model to Study Epithelial–Stromal Interactions in the Gut

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