eoe sub articles

EoE Sub Articles

1. Functional Characterization of Acquired Docetaxel Resistance by Colony Formation Assay
NOTE: In this protocol, chemoresistance has been evaluated using colony formation assays. Alternative methods used to evaluate cell viability (i.e.,&#160;MTS assays) can be used based on investigators&#39; preferences.
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	<li>Plate 2,000 cells (DU145 or 22Rv1, both parental and Docetaxel-resistant) per well in 6-well plates, using 2 mL of media per w...

functional chemoresistance characterization: a method to evaluate drug resistance in cancer cells using clonogenic assay

Source: Mohr, L. et al. <a href="https://www.jove.com/video/56327" target="_blank">Generation of Prostate Cancer Cell Models of Resistance to the Anti-mitotic Agent Docetaxel.</a> J. Vis. Exp. (2017)
This video describes a method for evaluating chemotherapeutic drug resistance in prostate cancer cells using colony formation assay. This assay can be used as a functional tool to identify novel molecular and cellular mechanisms of chemoresistance in cancer cells.

<img alt="Figure 1" src="/files/ftp_upload/20400/20400_Figure1.jpg" /> 
Figure 1: Functional and phenotypic characterization of the Docetaxel-resistant cell models.&#160;(A) Representative colony formation assays of parental and Docetaxel-resistant cells treated with the indicated Docetaxel concentrations for 72 h. Percentage of colonies for every treatment concentration is represented in the included graph. The experiments are triplicates and data r...

Functional Chemoresistance Characterization: A Method to Evaluate Drug Resistance in Cancer Cells Using Clonogenic Assay

Source: Mohr, L. et al. Generation of Prostate Cancer Cell Models of Resistance to the Anti-mitotic Agent Docetaxel. J. Vis. Exp. (2017)
This video ...

To evaluate chemoresistance in cancer cells, begin by taking a multi-well plate. Seed a suspension of the desired drug-sensitive prostate cancer cells into a few wells and a suspension of the drug-resistant variant of the cells into the remaining wells.
Incubate the culture to facilitate the attachment of cells to the culture plate. Treat both the cell types with a range of increasing doses of the selected drug solution. The drug molecules enter the sensitive cells and cause cell death even at their lowest concentrations. However, the resistant cells expel the drug molecules through specialized drug-efflux pumps, enabling their survival despite being exposed to higher drug concentrations.
Aspirate the spent media containing debris and drug solution. Supplement with fresh drug-free growth media. Incubate to allow the surviving drug-resistant cells to proliferate and form colonies. Add a suitable staining solution to color the live-cell colonies.
Wash the culture to remove excess dye. Count the number of colored colonies in each well. The wells containing drug-sensitive cells hold very few viable colonies. In contrast, the wells containing drug-resistant cells exhibit a higher number of colonies. However, these drug-resistant cells display a reduction in the number of colonies with increasing drug doses.

Functional Chemoresistance Characterization: A Method to Evaluate Drug Resistance in Cancer Cells Using Clonogenic Assay

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