Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/161945
Title: Development of reconstructed intestinal micronucleus cytome (RICyt) assay in 3D human gut model for genotoxicity assessment of orally ingested substances
Authors: Lim, Hui Kheng
Hughes, Christopher Owen
Lim, Michelle Jing Sin
Li, Jia'En Jasmine
Rakshit, Moumita
Yeo, Calvin
Chng, Kern Rei
Li, Angela
Chan, Joanne Sheot Harn
Ng, Kee Woei
Leavesley, David Ian
Smith, Benjamin Paul Chapman
Keywords: Engineering::Materials
Issue Date: 2022
Source: Lim, H. K., Hughes, C. O., Lim, M. J. S., Li, J. J., Rakshit, M., Yeo, C., Chng, K. R., Li, A., Chan, J. S. H., Ng, K. W., Leavesley, D. I. & Smith, B. P. C. (2022). Development of reconstructed intestinal micronucleus cytome (RICyt) assay in 3D human gut model for genotoxicity assessment of orally ingested substances. Archives of Toxicology, 96(5), 1455-1471. https://dx.doi.org/10.1007/s00204-022-03228-y
Project: IAF-PP-H18/01/a0/G14 
W20W3D0002 
Journal: Archives of Toxicology 
Abstract: The micronucleus (MN) assay is widely used as part of a battery of tests applied to evaluate the genotoxic potential of chemicals, including new food additives and novel food ingredients. Micronucleus assays typically utilise homogenous in vitro cell lines which poorly recapitulate the physiology, biochemistry and genomic events in the gut, the site of first contact for ingested materials. Here we have adapted and validated the MN endpoint assay protocol for use with complex 3D reconstructed intestinal microtissues; we have named this new protocol the reconstructed intestine micronucleus cytome (RICyt) assay. Our data suggest the commercial 3D microtissues replicate the physiological, biochemical and genomic responses of native human small intestine to exogenous compounds. Tissues were shown to maintain log-phase proliferation throughout the period of exposure and expressed low background MN. Analysis using the RICyt assay protocol revealed the presence of diverse cell types and nuclear anomalies (cytome) in addition to MN, indicating evidence for comprehensive DNA damage and mode(s) of cell death reported by the assay. The assay correctly identified and discriminated direct-acting clastogen, aneugen and clastogen requiring exogenous metabolic activation, and a non-genotoxic chemical. We are confident that the genotoxic response in the 3D microtissues more closely resembles the native tissues due to the inherent tissue architecture, surface area, barrier effects and tissue matrix interactions. This proof-of-concept study highlights the RICyt MN cytome assay in 3D reconstructed intestinal microtissues is a promising tool for applications in predictive toxicology.
URI: https://hdl.handle.net/10356/161945
ISSN: 0340-5761
DOI: 10.1007/s00204-022-03228-y
Rights: © 2022 The Author(s). Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:MSE Journal Articles
NEWRI Journal Articles

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