Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity

Nanoparticles are increasingly applied in the food industry without the availability of risk assessments. To reduce animal testing in risk assessments, improvements to in vitro testing have been made over the years. Here, we aim to develop a co-culture system using 3D intestinal model and TK6 cells...

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Main Author: Lim, Michelle Jing Sin
Other Authors: David Leavesley
Format: Final Year Project
Language:English
Published: 2019
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Online Access:http://hdl.handle.net/10356/79004
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-790042023-02-28T18:05:32Z Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity Lim, Michelle Jing Sin David Leavesley School of Biological Sciences Skin Research Institute of Singapore Science::Biological sciences Nanoparticles are increasingly applied in the food industry without the availability of risk assessments. To reduce animal testing in risk assessments, improvements to in vitro testing have been made over the years. Here, we aim to develop a co-culture system using 3D intestinal model and TK6 cells to study food nanogenotoxicity using cytokinesis-blocked micronucleus assay. As both cultures are grown with different media, optimisation was done to cultivate both tissue and cells in the same media to minimize changes to their native characteristics. RPMI, SMI, 10:90 and 25:75 RPMI:SMI media composition were tested on 3D tissues for 4 days and TK6 for 11 days. Using real-time PCR, both cultures had slower proliferation and were less inflamed, apoptotic and necrotic when grown in media other than their base media. For TK6, a minor and insignificant increase in the population doubling time was observed in media with SMI after 4 culture days. No changes to the basal micronuclei level in binucleated cells grown in different media compositions was seen. Furthermore, cytostasis increased throughout the culture period in all the media composition, with TK6 in SMI having the highest level of inhibition. Collectively, the results point to using SMI as the co-culture media. Bachelor of Science in Biological Sciences 2019-11-25T01:19:37Z 2019-11-25T01:19:37Z 2019 Final Year Project (FYP) http://hdl.handle.net/10356/79004 en Nanyang Technological University 39 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
spellingShingle Science::Biological sciences
Lim, Michelle Jing Sin
Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity
description Nanoparticles are increasingly applied in the food industry without the availability of risk assessments. To reduce animal testing in risk assessments, improvements to in vitro testing have been made over the years. Here, we aim to develop a co-culture system using 3D intestinal model and TK6 cells to study food nanogenotoxicity using cytokinesis-blocked micronucleus assay. As both cultures are grown with different media, optimisation was done to cultivate both tissue and cells in the same media to minimize changes to their native characteristics. RPMI, SMI, 10:90 and 25:75 RPMI:SMI media composition were tested on 3D tissues for 4 days and TK6 for 11 days. Using real-time PCR, both cultures had slower proliferation and were less inflamed, apoptotic and necrotic when grown in media other than their base media. For TK6, a minor and insignificant increase in the population doubling time was observed in media with SMI after 4 culture days. No changes to the basal micronuclei level in binucleated cells grown in different media compositions was seen. Furthermore, cytostasis increased throughout the culture period in all the media composition, with TK6 in SMI having the highest level of inhibition. Collectively, the results point to using SMI as the co-culture media.
author2 David Leavesley
author_facet David Leavesley
Lim, Michelle Jing Sin
format Final Year Project
author Lim, Michelle Jing Sin
author_sort Lim, Michelle Jing Sin
title Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity
title_short Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity
title_full Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity
title_fullStr Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity
title_full_unstemmed Optimisation of 3D intestine model & TK6 co-culture media to aid in the development of 3D micronuclei assay to assess Food nanogenotoxicity
title_sort optimisation of 3d intestine model & tk6 co-culture media to aid in the development of 3d micronuclei assay to assess food nanogenotoxicity
publishDate 2019
url http://hdl.handle.net/10356/79004
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