We are conducting research on early stage embryos and have twice now received a bad batch of media from our commercial supplier. This is extremely costly in time and funds. I am wondering if there is an analytic method to testing the quality of the media. I know the most obvious answer is to QC with "test" embryos but those are not readily available. Is there a way to check the composition via hplc, MS, ect to come up with a qualitative analysis compared against a "good" batch of media?
Cheers!
well the obvious answer is that this is entirely the problem of the supplier. They are responsible for the quality control of their product. I'd contact them and start a compliant, request that they do an extensive QC analysis and find out what the problem is. Also, I'd ask them for compensation. I have had this in the past with producers and these were always very willing to share info with me, as well as compensate for losses caused by their product.
The problem is that you do not know what to look for: osmolality, pH, etc are obvious, but endotoxins, soluble vitamins, amino acids, carbohydrates, contamination during production from the various unit operations etc are a different cookie...besides, testing may turn out to be labour intensive and expensive.
You could use appropriate cell lines to check for cell viability and proliferation. This is done for QC testing in chick embryo media (cell lines L929, HEL 299)
If you have some line culture cells, you could do a quick LDH test to look for cell death. You can buy the kits through Roche and others. Takes less than an hour. You can also do WST-1 or even just MTT to look for cell viability.
Thanks for the input. I agree, the easiest measure to take would be simply doing viability testing on an alternate cell culture. I was simply curious if it would be possible to create a media profile of a good batch to set as a standard to compare new incoming lots against.
In doing some reading I came across this:
Ryan, Paul W., et al. "Prediction of cell culture media performance using fluorescence spectroscopy." Analytical chemistry 82.4 (2010): 1311-1317.
seems a bit involved though.
well the paper you looked at is interesting, but it is a minimal quality methid, because it is based on the detection of components that fluoresce, i.e., the amino acids in proteins that fluoresce (tryptophan, tyrosine, phenylalanine), and cofactors such as FMN, FAD, NAD. It does not assess non-fluorescent components. This type of FL spectroscopy is also a bit elaborate to do.
I'd stick with a viability test and cell lines.
Your idea regarding HPLC-MS was a good one though (could make a fingerprint of the medium), but is rather elaborate and expensive and requires specialists
When culturing embryos, there are more potential toxic components than just culture media. In fact, culture media without protein should be the least likely concern. We've published a comprehensive review of culture media used for human embryo culture. Testing the components would be cost prohibitive and in my opinion, would not likely provide the answer.
Which media did you use? Was it already supplemented with protein? We have seen toxicity with mineral oil and with protein supplements used for embryo culture - these are both very heterogeneous and compared to culture media, poorly defined.
Though we have the capability to do so, we still use a bioassay of one-cell mouse embryos for verifying the quality of the products prior to use. While we all would like to put this on the manufacturer to provide a quality product, the undefined nature of the components like oil and protein, combined with different conditions used for culture (e.g. 20 vs 5% oxygen) by the end user and other unkonwn interactions make it unlikely they can detect all possible toxins.
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