eoe sub articles

EoE Sub Articles

1. Generation of Docetaxel-resistant Prostate Cancer Cells
NOTE: Perform cell treatments using Docetaxel solution stock used for the determination of IC50 drug concentration (prepare enough stock in advance for experimental use). Parental cells should be grown in parallel in a small flask throughout the whole procedure and exposed to vehicle (DMSO) in corresponding volumes mimicking the amounts used in the Docetaxel treated flasks.
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	<li>Plate DU145 or 2...

generating chemoresistant prostate cancer cells: a procedure for obtaining drug-resistant cancer cells in vitro

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 protocol for generating taxane-resistant cell models of prostate cancer. These models can help study the pathways involved in progression to Docetaxel resistance in prostate cancer patients.

<img alt="Figure 1" src="/files/ftp_upload/20399/20399_Figure1.jpg" /> 
Figure 1: Generation of Docetaxel-resistant prostate cancer cell model systems.&#160;(A) Diagram depicting the determination of the Docetaxel IC50 in the parental prostate cancer cell lines DU145 and 22Rv1 (Step 1 of the protocol). Expected percentages of cell viability and the Docetaxel dose-escalating concentrations needed are included. Representative DU145 and 22Rv1 cell viabi...

Generating Chemoresistant Prostate Cancer Cells: A Procedure for Obtaining Drug-resistant Cancer Cells In Vitro

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 generate chemoresistant cells, begin by taking human prostate cancer cell suspension in a culture flask. Incubate the flask to facilitate cell adherence and their division to form a monolayer.
Supplement the cells with an anti-cancer drug solution at an optimum concentration and incubate. The drug molecules enter the cells and induce apoptosis in sensitive cells.
Some cells express special drug efflux transporters or pumps that periodically remove the drugs. This mechanism prevents drug accumulation, enabling these cells to withstand the effect of anti-cancer drugs and become chemoresistant.
Aspirate the drug- and debris-containing media from the culture. Supplement with fresh drug-free growth media to support the growth of chemoresistant cells.&#160; Discard the spent media. Trypsinize the cells to detach them from the culture flask and transfer them to a tube.
Centrifuge the tube to pelletize the cells. Remove the trypsin-containing supernatant. Resuspend the cells in fresh growth media. Grow the cells in a new culture flask.
Subsequently, repeat the drug treatment and cell culture steps to expose the chemoresistant cells to escalating doses of the same anti-cancer drug. This technique screens the culture by eliminating all the sensitive cells to generate a pool of highly chemoresistant surviving cells.

Generating Chemoresistant Prostate Cancer Cells: A Procedure for Obtaining Drug-resistant Cancer Cells In Vitro

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