Its common to work with plant samples over various cancer cells. I saw many articles reported cytotoxicity at high extract concentrations. However, that cytotoxicity may be due to the concentration effect. Normal cell line of that cancer is maybe an option to work with. If normal cell line not available, what else?
Firstly, I would like to know if you would like to check if the plant extract has lethality towards the cancer cells or not.? If so the conventional method is preparation of varied dilutions of the plant extract and perform MTT assay on stable populated cancer cells and determine the minimum Inhibitory concentration. This will give a clear idea about which possible concentration is showing cytotoxicity. As any thing in excess is poision, an increased concentration of plant extract may be lethal to even normal cells. So purification and isolation of compounds/cell components within the extract is necessary to narrow down which is the key molecule to show inhibition.
Now if one wants to exclude the effect of cell death due to higher concentrations of plant extract, One can extract different cell fractions from the obtained plant extract like effect of total protein on cancer cells/ effect of fragmented DNA on cancer cells/ or any other fraction.
And as u have mentioned IF NORMAL CELL LINE NOT AVAILABLE. I dnt think one cannot get a cancer cell line of A type if the normal cell of that particular A type is present.
I hope my answer was justifying your question.
You have to check the toxicity of your compound in normal cells must. It is compulsory. You should buy any kind of normal cells like HUVEC, HEK, 3T3, CHO etc.
I think it depends on what you are trying to portray and what you are trying to ultimately achieve. Ill start off by saying what I would like to achieve
If you want to identify the active constituent in the extract, the maximum tolerated dose of the extract doesnt matter, rather only that of the active compound. Instead fractionate your extract by some chromatography method (silica or C18) then screen fractionated extracts, then find active compound and get an EC/IC50 value for the active compound.
If you want to screen the extract and are not worried about identifying the active constituent, then just screen your extract at a concentration starting at about 10 mg/ml and do 2 fold dilutions (maybe 8 or 16 consecutive 2x dilutions) and screen that against your cell line of interest. Be aware that different cancer cell lines will have different EC/IC50 values to your extract and unfortunately there is no good way to standardize this. So, what you could do is screen a known compound at the same time you screen your extract in the same dilution method mentioned above. The standards EC/IC50 value would help give your extract some point of reference for other people to compare.
Good luck!
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