Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production
Background and Aims: Necrotizing enterocolitis is a severe gastrointestinal complication of prematurity. Using small intestinal organoids derived from fetal tissue of a gestational age similar to an extremely preterm infant, this study aims to assess the effect of diet on intestinal epithelial growt...
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Elsevier
2024-01-01
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| Series: | Gastro Hep Advances |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S277257232400102X |
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| author | Lauren Smith Eduardo Gonzalez Santiago Chino Eke Weihong Gu Wenjia Wang Dhana Llivichuzhca-Loja Tessa Kehoe Kerri St Denis Madison Strine Sarah Taylor George Tseng Liza Konnikova |
| author_facet | Lauren Smith Eduardo Gonzalez Santiago Chino Eke Weihong Gu Wenjia Wang Dhana Llivichuzhca-Loja Tessa Kehoe Kerri St Denis Madison Strine Sarah Taylor George Tseng Liza Konnikova |
| author_sort | Lauren Smith |
| collection | DOAJ |
| description | Background and Aims: Necrotizing enterocolitis is a severe gastrointestinal complication of prematurity. Using small intestinal organoids derived from fetal tissue of a gestational age similar to an extremely preterm infant, this study aims to assess the effect of diet on intestinal epithelial growth and differentiation to elucidate the role nutrition type plays in intestinal development and modifies the risk for necrotizing enterocolitis. Methods: Organoids were cultured for 5 days in growth media and 5 days in differentiation media supplemented 1:40 with 4 different diets: parental milk, donor human milk, standard formula, or extensively hydrolyzed formula. Images were captured daily and organoids were quantified. Organoids were preserved for RNA sequencing and immunofluorescence staining with Ki67, cleaved caspase 3, and chromogranin-A. Media was saved for cytokine/chemokine and growth factor analysis. Results: Human milk supplementation improved growth and differentiation of intestinal organoids generating larger organoids during the growth phase and organoids with longer and wider buds during differentiation compared to formula. Ki67 staining confirmed the proliferative nature of milk-supplemented organoids and chromogranin A staining proved that MM-supplemented organoids induced highest enteroendocrine differentiation. Human milk supplementation also upregulated genes involved in Wnt signaling and fatty acid metabolism pathways and promoted a homeostatic immune landscape, including via increased secretion of tumor necrosis factor-related apoptosis-inducing ligand among other cytokines. Conversely, organoids supplemented with formula had a downregulation of cell-cycle-promoting genes and a more inflammatory immune signature, including a reduced level of leukemia inhibitory factor. Conclusion: Our results demonstrate that parental milk, and to a lesser extent donor human milk, support robust intestinal epithelial proliferation, differentiation, and homeostatic cytokine production, suggesting a critical role for factors enriched in human milk in intestinal epithelial health. |
| format | Article |
| id | doaj-art-1073bbd3b34244c0bcb698773f4c1f4e |
| institution | Kabale University |
| issn | 2772-5723 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Gastro Hep Advances |
| spelling | doaj-art-1073bbd3b34244c0bcb698773f4c1f4e2024-11-29T06:26:23ZengElsevierGastro Hep Advances2772-57232024-01-013810301042Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine ProductionLauren Smith0Eduardo Gonzalez Santiago1Chino Eke2Weihong Gu3Wenjia Wang4Dhana Llivichuzhca-Loja5Tessa Kehoe6Kerri St Denis7Madison Strine8Sarah Taylor9George Tseng10Liza Konnikova11Department of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Biostatistics, University of Pittsburgh, Pittsburgh, PennsylvaniaDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Pediatrics, Yale School of Medicine, New Haven, ConnecticutDepartment of Biostatistics, University of Pittsburgh, Pittsburgh, PennsylvaniaDepartment of Pediatrics, Yale School of Medicine, New Haven, Connecticut; Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, Connecticut; Department of Immunobiology, Yale School of Medicine, New Haven, Connecticut; Program in Human and Translational Immunology, Yale School of Medicine, New Haven, Connecticut; Program in Translational Biomedicine, Yale School of Medicine, New Haven, Connecticut; Center for Systems and Engineering Immunology, Yale School of Medicine, New Haven, Connecticut; Correspondence: Address correspondence to: Liza Konnikova, MD, PhD, Division of Neonatal-Perinatal Medicine, Department of Pediatrics, Yale School of Medicine, 375 Congress Ave, LSOG 405B, New Haven, Connecticut 06519.Background and Aims: Necrotizing enterocolitis is a severe gastrointestinal complication of prematurity. Using small intestinal organoids derived from fetal tissue of a gestational age similar to an extremely preterm infant, this study aims to assess the effect of diet on intestinal epithelial growth and differentiation to elucidate the role nutrition type plays in intestinal development and modifies the risk for necrotizing enterocolitis. Methods: Organoids were cultured for 5 days in growth media and 5 days in differentiation media supplemented 1:40 with 4 different diets: parental milk, donor human milk, standard formula, or extensively hydrolyzed formula. Images were captured daily and organoids were quantified. Organoids were preserved for RNA sequencing and immunofluorescence staining with Ki67, cleaved caspase 3, and chromogranin-A. Media was saved for cytokine/chemokine and growth factor analysis. Results: Human milk supplementation improved growth and differentiation of intestinal organoids generating larger organoids during the growth phase and organoids with longer and wider buds during differentiation compared to formula. Ki67 staining confirmed the proliferative nature of milk-supplemented organoids and chromogranin A staining proved that MM-supplemented organoids induced highest enteroendocrine differentiation. Human milk supplementation also upregulated genes involved in Wnt signaling and fatty acid metabolism pathways and promoted a homeostatic immune landscape, including via increased secretion of tumor necrosis factor-related apoptosis-inducing ligand among other cytokines. Conversely, organoids supplemented with formula had a downregulation of cell-cycle-promoting genes and a more inflammatory immune signature, including a reduced level of leukemia inhibitory factor. Conclusion: Our results demonstrate that parental milk, and to a lesser extent donor human milk, support robust intestinal epithelial proliferation, differentiation, and homeostatic cytokine production, suggesting a critical role for factors enriched in human milk in intestinal epithelial health.http://www.sciencedirect.com/science/article/pii/S277257232400102XIntestinal DevelopmentBreast MilkNecrotizing Enterocolitis |
| spellingShingle | Lauren Smith Eduardo Gonzalez Santiago Chino Eke Weihong Gu Wenjia Wang Dhana Llivichuzhca-Loja Tessa Kehoe Kerri St Denis Madison Strine Sarah Taylor George Tseng Liza Konnikova Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production Gastro Hep Advances Intestinal Development Breast Milk Necrotizing Enterocolitis |
| title | Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production |
| title_full | Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production |
| title_fullStr | Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production |
| title_full_unstemmed | Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production |
| title_short | Human Milk Supports Robust Intestinal Organoid Growth, Differentiation, and Homeostatic Cytokine Production |
| title_sort | human milk supports robust intestinal organoid growth differentiation and homeostatic cytokine production |
| topic | Intestinal Development Breast Milk Necrotizing Enterocolitis |
| url | http://www.sciencedirect.com/science/article/pii/S277257232400102X |
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