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<p>The development of a reliable ex vivo intestinal model is essential for studying gut physiology and host–microbe interactions under controlled conditions. This study evaluated the temporal dynamics of viability, morphology, and apoptotic regulation in chicken ileal explants cultured ex vivo. Ileal tissues from 21-d-old broiler chickens were incubated for 0, 4, and 8&#xa0;h and analyzed using metabolic assays (MTS, CCK-8), histological staining (hematoxylin and eosin (H&amp;E), Periodic Acid–Schiff (PAS)), and quantitative PCR for apoptosis-related genes (BCL2L11, cytochrome c, caspase-3, caspase-8, and caspase-9). After 4&#xa0;h, explants retained structural integrity with well-developed villi and active mucus secretion. Metabolic assays confirmed preserved cell viability, while transcriptional analysis revealed moderate upregulation of cytochrome c and downregulation of BCL2L11, suggesting transient adaptation to culture stress. After 8&#xa0;h, villi appeared shortened with epithelial exfoliation and reduced PAS reactivity, indicating progressive tissue deterioration. Correspondingly, BCL2L11 and cytochrome c were significantly upregulated together with caspase-8, reflecting activation of intrinsic and extrinsic apoptotic pathways. Metabolic activity declined markedly, confirming reduced tissue vitality. These findings demonstrate that chicken ileal explants with a diameter of 4&#xa0;mm remain viable and functionally stable up to approximately 4&#xa0;h of ex vivo culture, after which apoptotic processes intensify, compromising tissue integrity. The results define the practical viability window for short-term intestinal explant studies and provide methodological guidance for future investigations on host–pathogen or probiotic interactions in poultry gut models.</p>

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Dynamics of viability and apoptotic regulation in chicken ileal explant culture

  • 期刊论文

  • 发表日期：

    2026年3月14日

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  • Zuzana Kiššová,
  • Klaudia Jaszcza,
  • Veronika Vinclérová,
  • Rege Anna Márton,
  • Máté Mackei,
  • Gábor Mátis,
  • Viera Karaffová

摘要

The development of a reliable ex vivo intestinal model is essential for studying gut physiology and host–microbe interactions under controlled conditions. This study evaluated the temporal dynamics of viability, morphology, and apoptotic regulation in chicken ileal explants cultured ex vivo. Ileal tissues from 21-d-old broiler chickens were incubated for 0, 4, and 8 h and analyzed using metabolic assays (MTS, CCK-8), histological staining (hematoxylin and eosin (H&E), Periodic Acid–Schiff (PAS)), and quantitative PCR for apoptosis-related genes (BCL2L11, cytochrome c, caspase-3, caspase-8, and caspase-9). After 4 h, explants retained structural integrity with well-developed villi and active mucus secretion. Metabolic assays confirmed preserved cell viability, while transcriptional analysis revealed moderate upregulation of cytochrome c and downregulation of BCL2L11, suggesting transient adaptation to culture stress. After 8 h, villi appeared shortened with epithelial exfoliation and reduced PAS reactivity, indicating progressive tissue deterioration. Correspondingly, BCL2L11 and cytochrome c were significantly upregulated together with caspase-8, reflecting activation of intrinsic and extrinsic apoptotic pathways. Metabolic activity declined markedly, confirming reduced tissue vitality. These findings demonstrate that chicken ileal explants with a diameter of 4 mm remain viable and functionally stable up to approximately 4 h of ex vivo culture, after which apoptotic processes intensify, compromising tissue integrity. The results define the practical viability window for short-term intestinal explant studies and provide methodological guidance for future investigations on host–pathogen or probiotic interactions in poultry gut models.