How do you make a drug-resistant cell line? determine IC50?

Discussion of all aspects of cellular structure, physiology and communication.

Moderators: Leonid, amiradm, BioTeam

Post Reply
curious1
Garter
Garter
Posts: 23
Joined: Mon May 15, 2006 4:45 pm

How do you make a drug-resistant cell line? determine IC50?

Post by curious1 » Tue Jan 30, 2007 6:29 pm

Hi all,

I am trying to study for chemotherapy resistance for two drugs. I was wondering if anyone knew the exact protocol for developing a drug resistant cell line. From the articles that I read they only say that drug resistant cell lines were produced from stepwise additions of the drug of interest but there was no detailed protocol. At what confluency is the drug initially added? Do we increase the drug dosage after every day and would we double the concentration each time? Also, I'm not sure how to determine the IC50 values for the cell lines. I did the MTT cell proliferation assay using different drug dilutions and obtained absorbance values from ELISA readings. However, I'm not quite sure what to make of the values in order to calculate the IC50 (the concentration of drug which inhibits cell growth by 50%). I have the values for untreated and drug treated cells how exactly do the absorbance values relate to percentage of cell growth, is there a formula to calculate this? Anyone have any idea?

Post Reply

Who is online

Users browsing this forum: No registered users and 3 guests