How should I prepare the sample of water to test its effects such as on diabetes in cell lines. I am using 3T3-L1 cells in this study. Do I need to extract the elements or compound from the water samples for example: caffeine from coffee, at first?
Do we need only to test the element not the water?
I am assuming that the purpose of your experiments is to study the effect of what is contained in the water sample in question, such as various minerals and other contaminants. It is not unreasonable to assume that some or many of the effects, if observed in your experimental model, will be concentration dependent.
If your study is related to only the original levels of contaminants present in the water at normal levels, you will still need to concentrate the contaminants by a method that may not alter the activity of contaminants. A safe method is to lyophilize a known volume to dryness (or a low volume), resuspend/dissolve what may be present in as little volume as possible , and add appropriate volume fraction of this material to your culture such that the resultant dilution is equivalent to 1x (what was originally present in water) or any other factor you are interested in testing.
Before adding water residue to cell culture, you will need to filter sterilize the concentrate or after adding it, you will filter the medium.
Curious about Zam Zam!
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Yes, you are right Mr. Tausif Alam. As I want to test the effects of that water sample to cell, do I need to concentrate at first the various element in that water? I would like to test the water sample directly to the cell, but surely it will cause DMEM to be diluted with water and cause not optimum result.
Do you have experience on using drug test on cell? Does the drug was been diluted first with water or directly added to the cell culture with desired dose?
Zam zam is still got lot of interesting studies to be discovered.. but I not testing it yet
Thank You
Upto 5-10% variation in volume is usually tolerated by cells, as long as you make up for the osmolarity by adding sufficient concentrated saline or phosphate buffer.
Alternatively, you can begin with powdered medium and constitute the material in suitably smaller volume to accommodate the addition of water portion to the medium.
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