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THE FREQUENT SAMPLING OF WOUND SCRATCH ASSAY REVEALS THE “OPPORTUNITY” WINDOW FOR QUANTITATIVE EVALUATION OF CELL MOTILITY-IMPEDING DRUGS

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dc.contributor.author Kauanova, Sholpan
dc.contributor.author Urazbayev, Arshat
dc.contributor.author Vorobjev, Ivan
dc.date.accessioned 2021-05-11T10:16:58Z
dc.date.available 2021-05-11T10:16:58Z
dc.date.issued 2021-03
dc.identifier.citation Kauanova, S., Urazbayev, A., & Vorobjev, I. (2021). The Frequent Sampling of Wound Scratch Assay Reveals the “Opportunity” Window for Quantitative Evaluation of Cell Motility-Impeding Drugs. Frontiers in Cell and Developmental Biology, 9. https://doi.org/10.3389/fcell.2021.640972 en_US
dc.identifier.issn https://www.frontiersin.org/articles/10.3389/fcell.2021.640972/full
dc.identifier.issn 2296-634X
dc.identifier.uri https://doi.org/10.3389/fcell.2021.640972
dc.identifier.uri http://nur.nu.edu.kz/handle/123456789/5380
dc.description.abstract Wound healing assay performed with automated microscopy is widely used in drug testing, cancer cell analysis, and similar approaches. It is easy to perform, and the results are reproducible. However, it is usually used as a semi-quantitative approach because of inefficient image segmentation in transmitted light microscopy. Recently, several algorithms for wound healing quantification were suggested, but none of them was tested on a large dataset. In the current study, we develop a pipeline allowing to achieve correct segmentation of the wound edges in >95% of pictures and extended statistical data processing to eliminate errors of cell culture artifacts. Using this tool, we collected data on wound healing dynamics of 10 cell lines with 10 min time resolution. We determine that the overall kinetics of wound healing is non-linear; however, all cell lines demonstrate linear wound closure dynamics in a 6-h window between the fifth and 12th hours after scratching. We next analyzed microtubule-inhibiting drugs’, nocodazole, vinorelbine, and Taxol, action on the kinetics of wound healing in the drug concentration-dependent way. Within this time window, the measurements of velocity of the cell edge allow the detection of statistically significant data when changes did not exceed 10–15%. All cell lines show decrease in the wound healing velocity at millimolar concentrations of microtubule inhibitors. However, dose-dependent response was cell line specific and drug specific. Cell motility was completely inhibited (edge velocity decreased 100%), while in others, it decreased only slightly (not more than 50%). Nanomolar doses (10–100 nM) of microtubule inhibitors in some cases even elevated cell motility. We speculate that anti-microtubule drugs might have specific effects on cell motility not related to the inhibition of the dynamic instability of microtubules. en_US
dc.language.iso en en_US
dc.publisher Frontiers Media en_US
dc.relation.ispartofseries Frontiers in Cell and Developmental Biology;9
dc.rights Attribution-NonCommercial-ShareAlike 3.0 United States *
dc.rights.uri http://creativecommons.org/licenses/by-nc-sa/3.0/us/ *
dc.subject wound healing en_US
dc.subject high-throughput en_US
dc.subject label-free microscopy en_US
dc.subject live-cell imaging en_US
dc.subject segmentation automation en_US
dc.subject cell motility en_US
dc.title THE FREQUENT SAMPLING OF WOUND SCRATCH ASSAY REVEALS THE “OPPORTUNITY” WINDOW FOR QUANTITATIVE EVALUATION OF CELL MOTILITY-IMPEDING DRUGS en_US
dc.type Article en_US
workflow.import.source science


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