Abstract
Self-organizing mini-intestines cultured ex vivo from intestinal biopsy/resected samples, termed intestinal organoids or enteroids, present a unique opportunity for mechanistic investigation of health and disease of the intestinal epithelium. These patient-derived epithelial cultures are nontransformed, retain the genetic background of the patient, maintain regional specificity, differentiate into all major cell types of the intestinal epithelium, and are physiologically active. The biological relevance of human intestinal enteroids also circumvents the need for animal models for studies on the human gastrointestinal epithelium. Coculture with human endogenous microbes allows for exciting new studies on microbial–host interactions.
While the popularity of organoids/enteroids for human research has risen drastically over the past decade, existing work and published methods are primarily limited to adult tissue. Here, we describe a concise and effective method for the establishment neonatal enteroids (including preterm and term) from surgically resected tissue or biopsy material. While the protocol works on adult tissue/biopsies, it has been specifically adopted and optimised for neonatal tissue. We detail the procedure at each stage ranging from human tissue collection and extraction of stem cells from the tissue, to passaging and general maintenance of organoid/enteroid lines, and how to freeze and revive lines as needed.
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Acknowledgments
This work was supported by the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Grant Agreement No. 794771 to C.J.S. and Public Health Service Grant U19AI116497 from the National Institutes of Health to M.K.E. We thank Xi-lei Zeng for her expertise and help with all enteroid cultures.
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Stewart, C.J., Estes, M.K., Ramani, S. (2020). Establishing Human Intestinal Enteroid/Organoid Lines from Preterm Infant and Adult Tissue. In: Amarnath, S. (eds) Innate Lymphoid Cells . Methods in Molecular Biology, vol 2121. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0338-3_16
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DOI: https://doi.org/10.1007/978-1-0716-0338-3_16
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0338-3
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