A Novel Use of Embryonic Gut Organoid Culture to Investigate Duodenal Atresia.

Autor: Jones MLM; F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia; Discipline of Surgery, Sydney Medical School, The University of Sydney, Sydney, NSW, Australia; Department of Paediatric Surgery, The Royal Children's Hospital, Melbourne, VIC, Australia. Electronic address: matthewlmjones@bigpond.com., Sarila G; F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia., O'Sullivan B; F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia; Department of Paediatric Surgery, The Royal Children's Hospital, Melbourne, VIC, Australia; Department of Paediatrics, The University of Melbourne, Melbourne, VIC, Australia., Haycock S; F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia; Department of Paediatric Surgery, The Royal Children's Hospital, Melbourne, VIC, Australia; Department of Paediatrics, The University of Melbourne, Melbourne, VIC, Australia., Chapuis P; Discipline of Surgery, Sydney Medical School, The University of Sydney, Sydney, NSW, Australia., King SK; F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia; Discipline of Surgery, Sydney Medical School, The University of Sydney, Sydney, NSW, Australia; Department of Paediatric Surgery, The Royal Children's Hospital, Melbourne, VIC, Australia; Department of Paediatrics, The University of Melbourne, Melbourne, VIC, Australia., Teague WJ; F. Douglas Stephens Surgical Research Laboratory, Murdoch Children's Research Institute, Melbourne, VIC, Australia; Discipline of Surgery, Sydney Medical School, The University of Sydney, Sydney, NSW, Australia; Department of Paediatric Surgery, The Royal Children's Hospital, Melbourne, VIC, Australia; Department of Paediatrics, The University of Melbourne, Melbourne, VIC, Australia.
Jazyk: angličtina
Zdroj: Journal of pediatric surgery [J Pediatr Surg] 2024 Oct; Vol. 59 (10), pp. 161611. Date of Electronic Publication: 2024 Jul 01.
DOI: 10.1016/j.jpedsurg.2024.06.020
Abstrakt: Background: The cause of duodenal atresia (DA) is not known. Tandler's "solid cord" hypothesis conflicts with current biological evidence. In humans, a genetic aetiology is supported by the association with Trisomy 21. Interruption of Fgf10 is the strongest genetic link to DA in mice, demonstrating an increased incidence and severity as embryos mature. This project aimed to develop an organoid model to facilitate ex vivo DA research on the FGF10/FGFR2b signalling pathway. We hypothesised that DA morphology represents an evolving spectrum of disease and that Fgf10 knockout organoids would vary in growth pattern compared to wild-type.
Methods: Organoids were cultured from the duodenum of E12.5 Fgf10 knockout, heterozygous and wild-type embryos, using an air-liquid interface with Growth Factor reduced Matrigel. Organoids were photographed every 48 h to observe growth. Organoids were isolated and fixed after 14 days, then stained with DAPI, KI-67, and cytokeratin to demonstrate proliferation and differentiation.
Results: Wild-type duodenum developed into crypt-forming organoids. Fgf10 heterozygous duodenum failed to progress beyond the development stage of spheroids. Fgf10 knockout duodenum failed to demonstrate any growth. Wholemount staining showed the greatest cell proliferation and differentiation in wild-type tissue.
Conclusion: This research presents a novel concept for the growth of embryonic gastrointestinal tissue to inform normal biology. The small sample numbers and restricted culture duration limit longer-term growth analysis. While this model serves as a potential ex vivo setting for future research, that research should consider organoid models with greater standardisation and other gastrointestinal regions.
Level of Evidence: Animal/laboratory study.
Competing Interests: Conflicts of interest The authors of this paper have no conflicts of interest to declare.
(Copyright © 2024 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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