Cargando…
Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors
Wound healing assay is a simple and cost-effective in vitro assay for assessing therapeutic impacts on cell migration. Its key limitation is the possible confoundment by other cellular phenotypes, causing misinterpretation of the experimental outcome. In this study, we attempted to address this prob...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6976598/ https://www.ncbi.nlm.nih.gov/pubmed/31969633 http://dx.doi.org/10.1038/s41598-020-57806-0 |
_version_ | 1783490337641070592 |
---|---|
author | Somchai, Parinyachat Phongkitkarun, Kriengkrai Kueanjinda, Patipark Jamnongsong, Supawan Vaeteewoottacharn, Kulthida Luvira, Vor Okada, Seiji Jirawatnotai, Siwanon Sampattavanich, Somponnat |
author_facet | Somchai, Parinyachat Phongkitkarun, Kriengkrai Kueanjinda, Patipark Jamnongsong, Supawan Vaeteewoottacharn, Kulthida Luvira, Vor Okada, Seiji Jirawatnotai, Siwanon Sampattavanich, Somponnat |
author_sort | Somchai, Parinyachat |
collection | PubMed |
description | Wound healing assay is a simple and cost-effective in vitro assay for assessing therapeutic impacts on cell migration. Its key limitation is the possible confoundment by other cellular phenotypes, causing misinterpretation of the experimental outcome. In this study, we attempted to address this problem by developing a simple analytical approach for scoring therapeutic influences on both cell migration and cell death, while normalizing the influence of cell growth using Mitomycin C pre-treatment. By carefully mapping the relationship between cell death and wound closure rate, contribution of cell death and cell migration on the observed wound closure delay can be quantitatively separated at all drug dosing. We showed that both intrinsic cell motility difference and extrinsic factors such as cell seeding density can significantly affect final interpretation of therapeutic impacts on cellular phenotypes. Such discrepancy can be rectified by using the actual wound closure time of each treatment condition for the calculation of phenotypic scores. Finally, we demonstrated a screen for strong pharmaceutical inhibitors of cell migration in cholangiocarcinoma cell lines. Our approach enables accurate scoring of both migrastatic and cytotoxic effects, and can be easily implemented for high-throughput drug screening. |
format | Online Article Text |
id | pubmed-6976598 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69765982020-01-29 Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors Somchai, Parinyachat Phongkitkarun, Kriengkrai Kueanjinda, Patipark Jamnongsong, Supawan Vaeteewoottacharn, Kulthida Luvira, Vor Okada, Seiji Jirawatnotai, Siwanon Sampattavanich, Somponnat Sci Rep Article Wound healing assay is a simple and cost-effective in vitro assay for assessing therapeutic impacts on cell migration. Its key limitation is the possible confoundment by other cellular phenotypes, causing misinterpretation of the experimental outcome. In this study, we attempted to address this problem by developing a simple analytical approach for scoring therapeutic influences on both cell migration and cell death, while normalizing the influence of cell growth using Mitomycin C pre-treatment. By carefully mapping the relationship between cell death and wound closure rate, contribution of cell death and cell migration on the observed wound closure delay can be quantitatively separated at all drug dosing. We showed that both intrinsic cell motility difference and extrinsic factors such as cell seeding density can significantly affect final interpretation of therapeutic impacts on cellular phenotypes. Such discrepancy can be rectified by using the actual wound closure time of each treatment condition for the calculation of phenotypic scores. Finally, we demonstrated a screen for strong pharmaceutical inhibitors of cell migration in cholangiocarcinoma cell lines. Our approach enables accurate scoring of both migrastatic and cytotoxic effects, and can be easily implemented for high-throughput drug screening. Nature Publishing Group UK 2020-01-22 /pmc/articles/PMC6976598/ /pubmed/31969633 http://dx.doi.org/10.1038/s41598-020-57806-0 Text en © The Author(s) 2020 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Somchai, Parinyachat Phongkitkarun, Kriengkrai Kueanjinda, Patipark Jamnongsong, Supawan Vaeteewoottacharn, Kulthida Luvira, Vor Okada, Seiji Jirawatnotai, Siwanon Sampattavanich, Somponnat Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors |
title | Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors |
title_full | Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors |
title_fullStr | Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors |
title_full_unstemmed | Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors |
title_short | Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors |
title_sort | novel analytical platform for robust identification of cell migration inhibitors |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6976598/ https://www.ncbi.nlm.nih.gov/pubmed/31969633 http://dx.doi.org/10.1038/s41598-020-57806-0 |
work_keys_str_mv | AT somchaiparinyachat novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT phongkitkarunkriengkrai novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT kueanjindapatipark novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT jamnongsongsupawan novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT vaeteewoottacharnkulthida novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT luviravor novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT okadaseiji novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT jirawatnotaisiwanon novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors AT sampattavanichsomponnat novelanalyticalplatformforrobustidentificationofcellmigrationinhibitors |