Hello, dear scientists
Can I seed cells with treating doses of the drug, and after that, add mtt reagent without removing media?
If I don't remove drug solution before adding mtt reagent, is it possible that drug solution interferes with mtt reagent?
Which one is better attaching with collagen or treating cells in suspension form at the first step?
Can collagen cause problems in the test?
Greetings,
Although simple and easily reproducible, MTT assay technique have its limitation with regards to interference from treatment component. From my experience, my desired compound in the absence of cells did reduce MTT solution to formazan. Therefore:
1) It would be better to do optimization of MTT assay with your treatment component or;
2) You could substitute MTT with other reagents. If you are looking for cheaper alternatives, perhaps you can consider trypan blue exclusion method or Alamar blue assay for suspension cells.
- I would remove media before adding MTT to ensure you have a properly timed controlled reaction
I replace with fresh medium before adding the reagent of the method CellTiter 96 Aqueous One Solution Cell Proliferation Assay (Promega).
Thanaletchumi Manogaran María Luisa Caballero Laurence Stuart Dawkins-Hall These questions are because of the problem that we don't have plate centrifuge in our lab. I can't change my test cause it relates to my thesis. I can attach my cells with collagen, but I'm not sure if the collagen can interfere with the process or not.
If you wish to see the effect of collagen on the suspension cells, then its better to attach the cells first. As you cannot change the assay, my suggestion is remove the drug solution before MTT assay to eliminate any chances of inaccuracy. Since collagen does not to react with MTT reagent, you would obtain result for suspension cells only.
p/s:
Asymmetric Collagen/chitosan Membrane Containing Minocycline-loaded Chitosan Nanoparticles for Guided Bone Regeneration
Hi Elham,
I agree with Thanaletchumi, and other thing, you could aspirate the medium with a pasteur pipette connected to vacuum, without a plate centrifuge.
María Luisa Caballero Thanaletchumi Manogaran many thanks, I will try your suggestions. Hope I get good results
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