Novel Analytical Platform For Robust Identification of Cell Migration Inhibitors
Autor: | Somponnat Sampattavanich, Siwanon Jirawatnotai, Kulthida Vaeteewoottacharn, Kriengkrai Phongkitkarun, Vor Luvira, Parinyachat Somchai, Supawan Jamnongsong, Seiji Okada, Patipark Kueanjinda |
---|---|
Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Programmed cell death
Mitomycin Drug Evaluation Preclinical Motility lcsh:Medicine Biology Article Cell Line Cell Movement Cytotoxic T cell Humans Cell migration lcsh:Science Cell Proliferation Wound Healing Multidisciplinary Cell Death Cell growth Mitomycin C lcsh:R High-throughput screening Phenotype Wide-field fluorescence microscopy Cell culture Cell Migration Inhibition Cancer research lcsh:Q Cell Migration Assays |
Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-12 (2020) |
ISSN: | 2045-2322 |
Popis: | 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. |
Databáze: | OpenAIRE |
Externí odkaz: |