Continuation? Kombucha Tea and MCF-7 - (Nov/17/2010 )

Hello,

I am a high school student looking for some guidance on her science fair project. I am looking to do a continuation on my project from last year, but do not really know where to go from here, so I was hoping for some advice from people who know their stuff.

My project last year was on the effects that Kombucha Tea had on MCF-7 cell proliferation. It was a very simple experiment in which I used different percentages of Kombucha Tea with Growth Medium and cultured MCF-7 cells for 7 days, then ran an MTT Assay. I repeated this with MRC-5 cells as a comparison. My results using this were all over the place, sometimes decreasing proliferation and at other times increasing proliferation.

I know that this was extremely simple, but I am looking to expand on this and somehow get more concrete results. Does anyone have any ideas of what I could do?

I had been recommended to somehow imitate what would happen to Kombucha Tea after being absorbed into the body, and then seeing it's effect on the cells, because the tea in a real-life situation would never fully come into contact with the cells. Is there some procedure that I could use to do this?

Thanks!

"Results all over the place" - do you mean differences between proliferation at different concentration or differences between cell lines?

One would expect to find some kind of relationship between concentration and proliferation - it may be linear, non-linear, threshold etc. If you're getting some kind of "scatter", for me it would indicate some problems with the experiment itself - eg. sample preparation, cell culture (passage no., infection, time of culture before and after administration of compound) or proliferation assay. It may be worth it to use some kind of compound and make standard curve for proliferation.

If you have differences between cell lines - it's not that surprising, different cell lines may have completely different physiological characteristics.

As for imitation of Kombucha Tea absorption into body - the most simple thing I can think of would be dialysis or ultrafiltration (both with low cut-off points). As for more complicated - PAMPA. Even more complicated - Caco-2 cell line transport experiment.

Thank you, what I meant was within the cell lines themselves. I only got to repeat the experiment three times, so it would happen that one time proliferation would increase, and the other it would decrease, and the third it would stay the same. Would it be worth it to repeat the experiment more times?

Thanks! I will definitely discuss those with my mentor.

That's a bit strange... (Which means you're doing something wrong - or you are a step from some major discovery... ) I would definitely repeat the experiment but not before meticulous check of all factors of your experiment - eg. sample, cells, culture condition, proliferation assay. Control samples are also very important - you can have cells which were non-treated (blank) or treated with a substance that stimulates them to proliferate more (positive) or less (negative) - they're important to show you if your cells are OK and assay is working properly. If your experiments are made in the different cell passage and results are in series that shows increase or decrease of sensitivity in time it may suggest some kind of cell line "ageing".

I'm not a cell biology expert, but IMO the results showing no definable relation between treatment and proliferation, may indicate that there is no effect, i.e. the finding that some increase and some decrease occurs, are at random. Anyway a control group is surely necessary to compare and analyse this better.
And the Kombucha Tea: I wonder how standardised the product is, i.e. different charges might contain different concentrations of the ingredients or even different ingredients at all, as it is a "natural" product made by fermenting tea with different yeasts and bacteria.

I did use a control group as my basis, and would calculate the proliferation based off of that.

What I considered could be the cause of it was that I made sure to use the same bottle of commercial tea every time, so maybe as it aged it's properties changed? Kombucha tea is so unstable and unknown, but I was just offered the opportunity to do some tests on it with bacteria to test mutations that it could cause, so it might actually give me some insight on this.