I was working on an enzyme that targets cancer cells. We are planning to encapuslate the enzyme with RBC (where the substrate of the enzyme can freely diffuse across membranes). However, I wonder how I can measure its effects in vitro on cancer cell lines.
I have read articles on directly using MTT assays, which they directly put RBC-enzymes on top of the cancer cells. However when i did this experiment, i have found several problems.
Firstly, the media is easily contaminated when I incubate for 3 days.
Secondly, somehow the growth of cancer cells is inhibited even when i add RBCs (without enzyme) only as a control. I tried 1-10x10^6 RBCs, and the more RBCs i add, the cancer cells are not growing as well as the negative control wells (cancer cells only).
What is the problems with the above setup?
Are there any methods to test the effects of RBC-enzyme in vitro?
Thanks.
hii Nathaniel,
First of all, MTT is not so reliable assay. Still it is first line of experiment which most of the researchers start their preliminary anti-cancer related work.
We recently completed encapsulation of MSCs work. we also looked its effect on cancer. We did Alamar blue assay. Also to performed Calcein-AM staining and measured florescence as well as IF images.
Even you can think for WST or Brdu assay say BrdU will be work for you. These are more sensitive than MTT.
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