Cargando…
Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro
Chimeric antigen receptor T (CAR-T) cells are cytotoxic T cells engineered to specifically kill cancer cells expressing specific target receptor(s). Prior CAR-T efficacy tests include CAR expression analysis by qPCR or ELISA, in vitro measurement of interferon-γ (IFNγ) or interleukin-2 (IL-2), and x...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9014740/ https://www.ncbi.nlm.nih.gov/pubmed/35441124 http://dx.doi.org/10.1093/abt/tbac008 |
_version_ | 1784688245910536192 |
---|---|
author | Wong, Kuan Un Shi, Jingxuan Li, Peng Wang, Haitao Jia, Yanwei Deng, Chuxia Jiang, Lianmei Wong, Ada Hang-Heng |
author_facet | Wong, Kuan Un Shi, Jingxuan Li, Peng Wang, Haitao Jia, Yanwei Deng, Chuxia Jiang, Lianmei Wong, Ada Hang-Heng |
author_sort | Wong, Kuan Un |
collection | PubMed |
description | Chimeric antigen receptor T (CAR-T) cells are cytotoxic T cells engineered to specifically kill cancer cells expressing specific target receptor(s). Prior CAR-T efficacy tests include CAR expression analysis by qPCR or ELISA, in vitro measurement of interferon-γ (IFNγ) or interleukin-2 (IL-2), and xenograft models. However, the in vitro measurements did not reflect CAR-T cytotoxicity, whereas xenograft models are low throughput and costly. Here, we presented a robust in vitro droplet microfluidic assay for CAR-T cytotoxicity assessment. This method not only enabled assessment of CAR-T cytotoxic activity under different fluid viscosity conditions, but also facilitated measurement of CAR-T expansion and dissection of mechanism of action via phenotype analysis in vitro. Furthermore, our data suggested that label-free cytotoxicity analysis is feasible by acquiring data before and after treatment. Hence, this study presented a novel in vitro method for assessment of cellular cytotoxicity that could potentially be applied to any cytotoxicity experiment with varying solvent composition. |
format | Online Article Text |
id | pubmed-9014740 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-90147402022-04-18 Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro Wong, Kuan Un Shi, Jingxuan Li, Peng Wang, Haitao Jia, Yanwei Deng, Chuxia Jiang, Lianmei Wong, Ada Hang-Heng Antib Ther Original Research Article Chimeric antigen receptor T (CAR-T) cells are cytotoxic T cells engineered to specifically kill cancer cells expressing specific target receptor(s). Prior CAR-T efficacy tests include CAR expression analysis by qPCR or ELISA, in vitro measurement of interferon-γ (IFNγ) or interleukin-2 (IL-2), and xenograft models. However, the in vitro measurements did not reflect CAR-T cytotoxicity, whereas xenograft models are low throughput and costly. Here, we presented a robust in vitro droplet microfluidic assay for CAR-T cytotoxicity assessment. This method not only enabled assessment of CAR-T cytotoxic activity under different fluid viscosity conditions, but also facilitated measurement of CAR-T expansion and dissection of mechanism of action via phenotype analysis in vitro. Furthermore, our data suggested that label-free cytotoxicity analysis is feasible by acquiring data before and after treatment. Hence, this study presented a novel in vitro method for assessment of cellular cytotoxicity that could potentially be applied to any cytotoxicity experiment with varying solvent composition. Oxford University Press 2022-03-24 /pmc/articles/PMC9014740/ /pubmed/35441124 http://dx.doi.org/10.1093/abt/tbac008 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Antibody Therapeutics. All rights reserved. For Permissions, please email: journals.permissions@oup.com https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Original Research Article Wong, Kuan Un Shi, Jingxuan Li, Peng Wang, Haitao Jia, Yanwei Deng, Chuxia Jiang, Lianmei Wong, Ada Hang-Heng Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro |
title | Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro |
title_full | Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro |
title_fullStr | Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro |
title_full_unstemmed | Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro |
title_short | Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro |
title_sort | assessment of chimeric antigen receptor t cytotoxicity by droplet microfluidics in vitro |
topic | Original Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9014740/ https://www.ncbi.nlm.nih.gov/pubmed/35441124 http://dx.doi.org/10.1093/abt/tbac008 |
work_keys_str_mv | AT wongkuanun assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT shijingxuan assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT lipeng assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT wanghaitao assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT jiayanwei assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT dengchuxia assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT jianglianmei assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro AT wongadahangheng assessmentofchimericantigenreceptortcytotoxicitybydropletmicrofluidicsinvitro |