How to make a drug resistant cell line?

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vlpinell
vlpinell's picture
How to make a drug resistant cell line?

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. I have the values for untreated and drug treated cells but I don't think that half the absorbance of the control equate to the IC50 does it? Anyone have any idea?

hilltrekker
hilltrekker's picture
Hi

Hi
Please check this paper out in this link, especially materials and methods.

ONCOLOGY REPORTS 17: 1163-1169, 2007

Biological comparison of ovarian cancer resistant cell lines to cisplatin and Taxol by two different administrations

spandidos-publications.com/serveFile/or_17_5_1163_PDF.pdf?...&item=PDF - 161k - Cached

Marusja
Marusja's picture
 Dear vlpinell,

 Dear vlpinell,

I found this letter of yours which is dated from 6/16/2009.
I am a PhD student now and I am working on drug resistance in lung cancer cells.
If I write my problem it would be written exactly the same way as it was yours three years before.
So, please could you contact me? I wander what kind of solution did you find then?

Thank you,
Sincerely,
Mariam

FardousElsenduny
FardousElsenduny's picture
Hi,

Hi,

You can have a look to the chapter (establishment of drug-resistant cell line) in the following book

Cancer Cell Culture Methods and Protocols
Edited by
Simon P. Langdon

I hope it will help you

good luck
Fardous

sameergupta1285
sameergupta1285's picture
It depends on whether you

It depends on whether you want to select your resistant cells by intermittent incremental dose or by pulse treatment.

Ana.siddique
Ana.siddique's picture
Hey Mariam,

Hey Mariam,

I found your post:

Dated: Marusja : 1/19/2013 7:06 AM
relating to 'vlpinell' query for establishing drug resistant cell lines. I'm a research fellow too and I'm working on drug resistance in lung cancer cells (NCI-H460). I was wondering if you could help me with this. Did you get any protocol for generating resistant lung cancer cell lines? Can you please share it with me? Also, i want to know with what anticancer drugs you're working or worked with? Help please.

Thank you,
Sincerely,
Ana