Hi.
I want to start experiments in a few months, but i want to cover every single detail before starting with such time consuming protocols.
To give a little bit of context, i aim to develop resistance to antiandrogens in breast cancer cell lines.
I´m already considering getting an incubator specifically for this task and working with several replicates.
Could you provide any recommendations in this regard? Anything from very specific to general, either way it will be highly appreciated.
Mauricio:
Before you start, consider that cancer is not a two-dimensional disease. You may be mislead by assuming that cell lines in 2D are relevant to cancer.
Good luck!
CS
Mauricio:
Before you start, consider that cancer is not a two-dimensional disease. You may be mislead by assuming that cell lines in 2D are relevant to cancer.
Good luck!
CS
I was developing tamoxifen resistant cells. It took about 7 months to get strong resistance: you have to see that your resistant cells grow well in the presence of the tamoxifen, that is very important - to get cells growing and dividing in the presence of the inhibitor, not just surviving, but growing. Another important point - to keep good positive control cells that die in the presence of your inhibitors. When you start to develop resistance , make sure to freeze the batch of cells that die very well after treatment with your inhibitors. Otherwise, you would not be able to compare the resistant cells with responsive controls.
Make sure that you get cells with good expression of androgen receptors - so you will be able to see the response to AR inhibitors. See the attached paper as reference for similar work. they were using AR antagonists enzalutamide and/or seviteronel. You might need to get AR agonists as well and compare the effects as additional positive controls. Good luck!
I forgot to add. You need to consider doing clonal selection. After your first treatment with an inhibitor, you can pool all survived cells, or collect separately several survived groups of cells and keep them growing separately. The collected clones might die, but some of them will survive, and at the end you will get several resistant cell lines. It is more expensive and more difficult work. I could not keep going clonal selection and ended up with pooled batch of resistant cells. I regretted it later when found out that it is a mixed population of very heterogeneous cells that show different characteristics. So, if you can get some help it is better to do clonal selection.
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