Cytotoxicity Assay Positive Control - (Jan/08/2007 )
I'm looking for a positive control for an MTT based cytotoxicity assay. I'm currently using a huh-7 liver cell line. Is there a standard chemical compound used as a cytotoxicity control? So far I haven't had any luck finding one. Thanks in advance for your help.
I am not sure abt liver cell line. For neurons or neuronal cell lines, people use glutamate, hydrogen peroxide, lactatic acid.
U could try hydrogen peroxide. How abt glucose deprivation ?
Good Luck !!!
Thanks for the suggestions scolix. I'm going to try them. Do you know of an approximate concentration people use for glutamate, lactic acid and hydrogen peroxide in neuronal cells to generate a cytotoxicity curve?
During my literature search today I came across Camptothecin, a DNA topoisomerase I inhibitor. According to a publication I found it has a CC50 of 13 uM in primary hepatocytes. I expect the dose may be higher in huh-7 cells. Have you or anyone else used it as a control before?
Thanks again for your help!
U could get the conc. in the literature as they vary depending on cell types. I have never used huh cells.
You might want to try 10 to 100 uM H202 for 30 min.
to induce cytotoxicity you may use vinblastin or staurosporin; you may like to compare your data to previously released data, so you have to adapt to the published conditions
I wanted to follow-up and say thanks to all that reply-ed to my post. It was quite helpful. Incase there is anyone interested in finding a cytotoxicity control for their assay in the future I have put my findings below for my assay. Thanks again!
Staurosporine: CC50 = 67 nM, CC90 = 280 nM
Camptothecin: CC50=297, CC90 --> never reached 100% cytotoxicity (Also, camptothecin was hard to soluablize in DMSO.)
Huh-7 cells (hepatocyte cell line)
Time with compound: 72 hours.