{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T15:29:39Z","timestamp":1781018979635,"version":"3.54.1"},"reference-count":18,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T00:00:00Z","timestamp":1764028800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004837","name":"Ministerio de Ciencia, Innovaci\u00f3n y Universidades","doi-asserted-by":"publisher","award":["PID2021-126489OB-I00"],"award-info":[{"award-number":["PID2021-126489OB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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 &lt; 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.<\/jats:p>","DOI":"10.3390\/ijms262311406","type":"journal-article","created":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T15:29:47Z","timestamp":1764084587000},"page":"11406","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["An Ovine Intestinal Organoid\u2013Macrophage Co-Culture Model to Test the Effects of Ovine Colostrum Exosomes on Intestinal Barrier Function and Inflammation"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3972-7293","authenticated-orcid":false,"given":"Mahsa","family":"Dehnavi","sequence":"first","affiliation":[{"name":"Instituto de Ganader\u00eda de Monta\u00f1a (CSIC-Universidad de Le\u00f3n), Finca Marzanas s\/n, E-24346 Grulleros, Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6854-5507","authenticated-orcid":false,"given":"Giulio","family":"Galli","sequence":"additional","affiliation":[{"name":"Departamento de Ciencias Biom\u00e9dicas, Facultad de Veterinaria, Universidad de Le\u00f3n, Campus de Vegazana s\/n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5617-9669","authenticated-orcid":false,"given":"Carlos","family":"Garc\u00eda-Estrada","sequence":"additional","affiliation":[{"name":"Departamento de Ciencias Biom\u00e9dicas, Facultad de Veterinaria, Universidad de Le\u00f3n, Campus de Vegazana s\/n, 24071 Le\u00f3n, Spain"},{"name":"Instituto de Biomedicina (IBIOMED), Universidad de Le\u00f3n, Campus de Vegazana s\/n, 24007 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0418-6116","authenticated-orcid":false,"given":"Rafael","family":"Bala\u00f1a-Fouce","sequence":"additional","affiliation":[{"name":"Departamento de Ciencias Biom\u00e9dicas, Facultad de Veterinaria, Universidad de Le\u00f3n, Campus de Vegazana s\/n, 24071 Le\u00f3n, Spain"},{"name":"Instituto de Biomedicina (IBIOMED), Universidad de Le\u00f3n, Campus de Vegazana s\/n, 24007 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0680-1541","authenticated-orcid":false,"given":"F. Javier","family":"Gir\u00e1ldez","sequence":"additional","affiliation":[{"name":"Instituto de Ganader\u00eda de Monta\u00f1a (CSIC-Universidad de Le\u00f3n), Finca Marzanas s\/n, E-24346 Grulleros, Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mercedes","family":"Alonso","sequence":"additional","affiliation":[{"name":"Instituto de Ganader\u00eda de Monta\u00f1a (CSIC-Universidad de Le\u00f3n), Finca Marzanas s\/n, E-24346 Grulleros, Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8802-3535","authenticated-orcid":false,"given":"Nuria","family":"Santos","sequence":"additional","affiliation":[{"name":"Instituto de Ganader\u00eda de Monta\u00f1a (CSIC-Universidad de Le\u00f3n), Finca Marzanas s\/n, E-24346 Grulleros, Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernando","family":"Rozada","sequence":"additional","affiliation":[{"name":"Instituto de Ganader\u00eda de Monta\u00f1a (CSIC-Universidad de Le\u00f3n), Finca Marzanas s\/n, E-24346 Grulleros, Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5255-1582","authenticated-orcid":false,"given":"Sonia","family":"Andr\u00e9s","sequence":"additional","affiliation":[{"name":"Instituto de Ganader\u00eda de Monta\u00f1a (CSIC-Universidad de Le\u00f3n), Finca Marzanas s\/n, E-24346 Grulleros, Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,25]]},"reference":[{"key":"ref_1","first-page":"686","article-title":"The intestinal barrier function and its involvement in digestive disease","volume":"107","author":"Vicario","year":"2015","journal-title":"Rev. Esp. Enferm. Dig."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4955","DOI":"10.3168\/jds.2015-10351","article-title":"Development and physiology of the rumen and the lower gut: Targets for improving gut health","volume":"99","author":"Steele","year":"2016","journal-title":"J. Dairy Sci."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Liu, W., Du, C., Nan, L., Li, C., Wang, H., Fan, Y., and Zhang, S. (2024). The difference of milk-derived extracellular vesicles from cow colostrum and mature milk on miRNAs expression and protecting intestinal epithelial cells against lipopolysaccharide damage. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25073880"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"116245","DOI":"10.1016\/j.anifeedsci.2025.116245","article-title":"Early post-natal methionine supplementation in ewe lambs: Long-term effects on feed efficiency, milk yield and fatty acid profile, metabolism, gut microbiome, and epigenetic regulation","volume":"321","author":"Dehnavi","year":"2025","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"116015","DOI":"10.1016\/j.anifeedsci.2024.116015","article-title":"Feed restriction during the suckling period of ewe Assaf lambs (F0) modifies milk quality and milk exosomal miRNAome of the filial generation (F1)","volume":"314","author":"Calogero","year":"2024","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"He, Y., He, Z., Leone, S., and Liu, S. (2021). Milk exosomes transfer oligosaccharides into macrophages to modulate immunity and attenuate adherent-invasive E. coli (AIEC) infection. Nutrients, 13.","DOI":"10.3390\/nu13093198"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lapping-Carr, G., Gemel, J., Mao, Y., and Beyer, E.C. (2020). Circulating extracellular vesicles and endothelial damage in sickle cell disease. Front. Physiol., 11.","DOI":"10.3389\/fphys.2020.01063"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Galli, G., Melc\u00f3n-Fern\u00e1ndez, E., de Garnica Garc\u00eda, M.G., Mart\u00ednez-Fern\u00e1ndez, B., Dehnavi, M., Andr\u00e9s, S., P\u00e9rez-Pertejo, Y., Reguera, R.M., Garc\u00eda-Estrada, C., and Mart\u00ednez-Valladares, M. (2025). Development of Sheep Duodenum Intestinal Organoids and Implementation of High-Throughput Screening Platform for Veterinary Applications. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26073452"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Miao, X., Luo, Q., Zhao, H., and Qin, X. (2017). An integrated analysis of miRNAs and methylated genes encoding mRNAs and lncRNAs in sheep breeds with different fecundity. Front. Physiol., 8.","DOI":"10.3389\/fphys.2017.01049"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Pokharel, K., Peippo, J., Honkatukia, M., Sepp\u00e4l\u00e4, A., Rautiainen, J., Ghanem, N., Hamama, T.M., Crowe, M.A., Andersson, M., and Li, M.H. (2018). Integrated ovarian mRNA and miRNA transcriptome profiling characterizes the genetic basis of prolificacy traits in sheep (Ovis aries). BMC Genom., 9.","DOI":"10.1186\/s12864-017-4400-4"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1186\/1476-4598-11-44","article-title":"Silencing of a large microRNA cluster on human chromosome 14q32 in melanoma: Biological effects of mir-376a and mir-376c on insulin growth factor 1 receptor","volume":"11","author":"Zehavi","year":"2012","journal-title":"Mol. Cancer"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1161\/ATVBAHA.108.181727","article-title":"Insulin-like growth factor-1 receptor expression masks the antiinflammatory and glucose uptake capacity of insulin in vascular smooth muscle cells","volume":"29","author":"Engberding","year":"2009","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Jin, M., Fei, X., Li, T., Lu, Z., Chu, M., Di, R., He, X., Wang, X., Wang, Y., and Yuan, Z. (2022). Oar-miR-432 regulates fat differentiation and promotes the expression of BMP2 in ovine preadipocytes. Front. Genet., 13.","DOI":"10.3389\/fgene.2022.844747"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Cui, Z., Amevor, F.K., Zhao, X., Mou, C., Pang, J., Peng, X., Liu, A., Lan, X., and Liu, L. (2023). Potential therapeutic effects of milk-derived exosomes on intestinal diseases. J. Nanobiotechnol., 21.","DOI":"10.1186\/s12951-023-02176-8"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Jab\u0142o\u0144ska, M., Sawicki, T., \u017bulewska, J., Staniewska, K., \u0141obacz, A., and Przyby\u0142owicz, K.E. (2024). The role of bovine Milk-Derived exosomes in human health and disease. Molecules, 29.","DOI":"10.3390\/molecules29245835"},{"key":"ref_16","first-page":"57","article-title":"Estimaci\u00f3n de las necesidades de amino\u00e1cidos en los corderos","volume":"7\u20138","author":"Cantarella","year":"1991","journal-title":"Mundo Ganad."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Nozi\u00e8re, P., Sauvant, D., and Delaby, L. (2018). INRA Feeding System for Ruminants, Wageningen Academic Publishers.","DOI":"10.3920\/978-90-8686-872-8"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"110131","DOI":"10.1016\/j.vetimm.2020.110131","article-title":"Optimized in vitro isolation of different subpopulation of immune cells from peripheral blood and comparative techniques for generation of monocyte-derived macrophages in small ruminants","volume":"230","author":"Benavides","year":"2020","journal-title":"Vet. Immunol. 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