Development of Sheep Duodenum Intestinal Organoids and Implementation of High-Throughput Screening Platform for Veterinary Applications DOI Open Access
Giulio Galli, Estela Melcón-Fernández, María Gracia de Garnica García

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(7), P. 3452 - 3452

Published: April 7, 2025

New therapeutic molecules for farm animals are needed to address worldwide problems in the food industry, like rise of resistance among ruminant parasites and pathogenic microbes. Since vivo testing would involve an excessive number animals, with consequent ethical economic issues, generation sheep intestinal organoids represents a promising close-to-reality vitro model veterinary drug development; however, characterization application such remain limited. In this study, ovine were generated from adult LGR5+ stem cells crypts freshly slaughtered lambs, developed culture system. Morphological analysis via brightfield microscopy immunocytochemical staining revealed pseudostratified epithelium multiple cell types, distinct apical-basal polarity, while RNA sequencing validated preservation physiological characteristics original organ. The development robust reproducible protocol culturing duodenum high-throughput screening (HTS) compatible format demonstrated reliability HTS applications, Z'-factor tests indicating assay performance. Dose-response studies using pre-identified compounds showed comparable pharmacodynamic profiles between mouse organoids. These findings establish as innovative tool pharmacology toxicology, offering cost-effective sustainable platform challenges improve livestock health.

Language: Английский

Development of Sheep Duodenum Intestinal Organoids and Implementation of High-Throughput Screening Platform for Veterinary Applications DOI Open Access
Giulio Galli, Estela Melcón-Fernández, María Gracia de Garnica García

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(7), P. 3452 - 3452

Published: April 7, 2025

New therapeutic molecules for farm animals are needed to address worldwide problems in the food industry, like rise of resistance among ruminant parasites and pathogenic microbes. Since vivo testing would involve an excessive number animals, with consequent ethical economic issues, generation sheep intestinal organoids represents a promising close-to-reality vitro model veterinary drug development; however, characterization application such remain limited. In this study, ovine were generated from adult LGR5+ stem cells crypts freshly slaughtered lambs, developed culture system. Morphological analysis via brightfield microscopy immunocytochemical staining revealed pseudostratified epithelium multiple cell types, distinct apical-basal polarity, while RNA sequencing validated preservation physiological characteristics original organ. The development robust reproducible protocol culturing duodenum high-throughput screening (HTS) compatible format demonstrated reliability HTS applications, Z'-factor tests indicating assay performance. Dose-response studies using pre-identified compounds showed comparable pharmacodynamic profiles between mouse organoids. These findings establish as innovative tool pharmacology toxicology, offering cost-effective sustainable platform challenges improve livestock health.

Language: Английский

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