" ...now we wants to measure toxicity of these cell line on normal cell line.." a mistake in your question ! Do you want to test the peptides on a normal cell line ?

What are the effect of the peptides on the cell cycle of the cancer cell lines ? Have you tested peptide cytotoxicity on resting (quiescent) cells ?

for checking the toxicity of molecules on cell lines, you need to choose appropriate cell line/s first followed by various toxicity assays. Some commonly used assays are LIVE/DEAD, LDH, Alamar blue, picogreen, etc. You can look up other assays in the literature. I would recommend using cancerous cell lines such as SKBR-3, MCF etc instead of others as your peptide has anti-cancerous activity.

We tested peptides on cancer cell lines and found that they are inhibiting growth of cancer cell lines. Now we are interested to know whether these peptides are toxic or not. I will appreciate if some body suggest protocol to measure toxicity of these peptides. Aim of this work is to identify anticancer peptides having minimum toxicity.

Hello

I also work on anticancer peptides. Basically you can test the cytotoxicity of peptides on normal cell lines like NIH3T3, L929 cells. Treat this cell lines with different concentrations of your peptides and then do MTT assay and trypan blue viability assay.

Hello Sir,

Any primary cells isolated from mice may be also a worthful model for testing these peptides may be.

If you need basic test for toxicity of your peptides with "well diffusion method" on bacterium. similarly assay is available for animal cell culture.

Hello Mr. Raghava,

First, I would suggest using NIH3T3 mouse embryonic fibroblasts and HEK-293 human kidney cells for normal cell lines. Also, I have conducted various assays to assess cytotoxicity of nanoparticles: including Alamar Blue, Presto Blue, Neutral Red, LDH, WST-1, Trypan Blue, and so on. These assays are all accurate, provided that some factors, in my case nanoparticles, do not interfere with absorbance and fluorescence measurements.

Lastly, for trypan blue, I was not able to distinguish between cells in the process of dying and cells that are healthy. Other than that, with any of the assays I have listed, you can vary the concentration of anticancer peptides and measure at different time points, just like any other cytotoxicity assay.

I am thankful to all of you for excellent suggestions. We will keep your suggestion in testing cytotoxicity of peptides.