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