I am working on Acute myeloid leukemia (AML) bone marrow derived Mesenchymal stem cells and will be studying their interactions with AML cells. For standardization process i need to work with drug resistant cell lines. Can anyone help me or suggest me Drug resistant AML cell lines for my study.
Hi Manasi,
What is the nature of your drugs? Are they well-characterized in the literature, and if so, there may be drug resistant cell lines already published by other groups that you can contact.
If these drugs have known molecular targets, you may want to consider knockdown or knockout of those targets in your cell line of choice to generate a "resistant" clone.
If these are novel drugs with no clear molecular targets or no such cell lines are available, you could consider generating your own resistant clones. Generally, this involves first calculating a dose response curve measuring viability or proliferation of your AML cells (e.g. HL60) with increasing drug concentration at multiple timepoints.
Then you'd culture your cells at a low drug concentration (for instance, enough to kill ~10% of your cells) for an extended period such as 7-10 doublings. You would continue escalating the drug concentration and continue passasing your cells. After about two months, you hopefully have a resistant population.
Some things to consider:
1) Do you know the stability of your drug in media? You may have to replenish the media depending on how fast the drug gets depleted or inactivated.
2) You may want to consider deriving clonal populations from your resistant cells (for instance 2-3 clones) to make sure your interactions are consistent across clones.
3) You may want to culture a non-drug selection flask in paralell with your drug selection flask so you can partly control for passaging numbers. That way, you should have by the end of selection both a drug senstive and drug resistance culture of similar passage.
I hope that clarifies your question.
Thanks,
Barry
You'd find protocols to induce drug resistance in the same cell line you are using. There could be some variations depending on the drug class and cells type but the main idea is always the same : exposure of the cell line to gradually increasing concentrations of the drug over a long time.
First of all, resistant against what?. Exist the multi-resistance phenomenon trigger for common mutations (usually TP53 and other strong drivers) but the most common studies want to decipher resistant against an specific drug of group of drugs (cytorreductors, alkylant agents, proteasome inhibitors...). Secondly, the best way to work with resistant cell line, once you know what is the drug or group of drugs is choose an specific cell line with the molecular profile (mutations carried by the cell line) that fits with your work, and develop a resistant cell line. Developing a resistant cell line its pretty simple, just time consuming. Basically put a selection pressure of the drug choosen during long-term. With the time, (1-12 months, it depends how quick proliferate your cell line) the Ic50 of your cell line againt the drug should be at least, doubled. When this happens, you can call your new cell line "resistant) (e.g. you start using OCI-AML3, and you start treating with trametinib, after 6 months OCI-AML3 raise from Ic50 of 10 uM to 25 uM; now you can claim that you create a new cell line, the OCI-AML3 Tramet. Resistant).
Hi Barry Zee,
Thank you so much for your suggestion. Well I am looking for AML cell lines that are resistant to conventional drugs cytarabine and etoposide for my study. My concern was to get a commercially available cell line resistant to the above mentioned drugs. I will be searching in the already published articles for such cell lines.
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