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Volume 26
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10.3390/ijms262311406
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Brief Report

An Ovine Intestinal Organoid–Macrophage Co-Culture Model to Test the Effects of Ovine Colostrum Exosomes on Intestinal Barrier Function and Inflammation

by
Mahsa Dehnavi
1,
Giulio Galli
2,
Carlos García-Estrada
2,3,
Rafael Balaña-Fouce
2,3,
F. Javier Giráldez
1,
Mercedes Alonso
1,
Nuria Santos
1,
Fernando Rozada
1 and
Sonia Andrés
1,*
1
Instituto de Ganadería de Montaña (CSIC-Universidad de León), Finca Marzanas s/n, E-24346 Grulleros, León, Spain
2
Departamento de Ciencias Biomédicas, Facultad de Veterinaria, Universidad de León, Campus de Vegazana s/n, 24071 León, Spain
3
Instituto de Biomedicina (IBIOMED), Universidad de León, Campus de Vegazana s/n, 24007 León, Spain
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2025, 26(23), 11406; https://doi.org/10.3390/ijms262311406
Submission received: 14 October 2025 / Revised: 19 November 2025 / Accepted: 21 November 2025 / Published: 25 November 2025
(This article belongs to the Special Issue Intestinal 3D Culture, Organoids and Gut-on-Chip in Intestinal Disorders)
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Abstract

Ovine colostrum exosomes obtained from nutritionally programmed dairy ewes (F0) may present modifications in microRNAs, thus having consequences for the intestinal barrier function and immunity parameters of lambs (F1). To test this hypothesis, colostrum exosomes from two ewe groups [F0-MET (nutritionally programmed ewes being fed methionine during early life) and F0-CTRL (ewes not supplemented with methionine during early life)] were sequenced to compare differences in the miRNAome. In addition, these exosomes were added to an in vitro co-culture in a Transwell chamber system consisting of ovine duodenum intestinal organoids and macrophages to assess the expression of genes encoding tight junction proteins in organoids and immunity parameters in macrophages. Finally, the concentrations of cytokines (e.g., IL-12 and IL-6) were assessed by ELISA kits in the supernatants of the chamber containing macrophages. According to the miRNAome, the expression of two miRNAs (e.g., oar_miR_376c_3p and oar_miR_432) was reduced in the colostrum exosomes obtained from dairy ewes nutritionally programmed with dietary supplementation of methionine during early life (F0-MET ewes). These changes did not seem to modify the expression of intestinal barrier and immune response marker genes when these exosomes were added to a co-culture of ovine intestinal organoids and macrophages. However, the levels of IL-12 produced by macrophages were reduced (p < 0.05), which suggests the inhibition of inflammatory pathways. Further studies using ovine colostrum exosomes obtained from nutritionally programmed ewes will help to clarify their potential to improve the health of suckling lambs.
Keywords: methionine; nutritional programming; three Rs; alternative methods; miRNA; epigenetics; organoids methionine; nutritional programming; three Rs; alternative methods; miRNA; epigenetics; organoids
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MDPI and ACS Style

Dehnavi, M.; Galli, G.; García-Estrada, C.; Balaña-Fouce, R.; Giráldez, F.J.; Alonso, M.; Santos, N.; Rozada, F.; Andrés, S. An Ovine Intestinal Organoid–Macrophage Co-Culture Model to Test the Effects of Ovine Colostrum Exosomes on Intestinal Barrier Function and Inflammation. Int. J. Mol. Sci. 2025, 26, 11406. https://doi.org/10.3390/ijms262311406

AMA Style

Dehnavi M, Galli G, García-Estrada C, Balaña-Fouce R, Giráldez FJ, Alonso M, Santos N, Rozada F, Andrés S. An Ovine Intestinal Organoid–Macrophage Co-Culture Model to Test the Effects of Ovine Colostrum Exosomes on Intestinal Barrier Function and Inflammation. International Journal of Molecular Sciences. 2025; 26(23):11406. https://doi.org/10.3390/ijms262311406

Chicago/Turabian Style

Dehnavi, Mahsa, Giulio Galli, Carlos García-Estrada, Rafael Balaña-Fouce, F. Javier Giráldez, Mercedes Alonso, Nuria Santos, Fernando Rozada, and Sonia Andrés. 2025. "An Ovine Intestinal Organoid–Macrophage Co-Culture Model to Test the Effects of Ovine Colostrum Exosomes on Intestinal Barrier Function and Inflammation" International Journal of Molecular Sciences 26, no. 23: 11406. https://doi.org/10.3390/ijms262311406

APA Style

Dehnavi, M., Galli, G., García-Estrada, C., Balaña-Fouce, R., Giráldez, F. J., Alonso, M., Santos, N., Rozada, F., & Andrés, S. (2025). An Ovine Intestinal Organoid–Macrophage Co-Culture Model to Test the Effects of Ovine Colostrum Exosomes on Intestinal Barrier Function and Inflammation. International Journal of Molecular Sciences, 26(23), 11406. https://doi.org/10.3390/ijms262311406

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