I am trying to make a custom synthesized media, where one of the constituents is human recombinant insulin. Can I substitute the commercial cell culture grade insulin (a bit costly) with the regular human insulin intended for sub-cutaneous injection. I would be grateful if some one share their expertise and experience in this.
Hi Raman, as far as I am aware you should be able to substitute the "diabetic" insulin for "cell culture insulin" (as both should bind and activate insulin receptors they should in theory be the same). If you wish you could easily check this by growing some target cells (such as MCF-7 cells) in media containing both insulin sources. By counting the cells in both conditions you should see if the insulin source has an effect or not (there should be no difference).
However I would still be tempted to purchase cell culture quality Insulin. As growth factors go it is very cheap (£39 for 5 ml of a 10 mg/ml solution).
http://www.sigmaaldrich.com/catalog/product/sigma/i9278?lang=en®ion=GB
For MCF-7 cells this is used at 1:1000 (0.01 mg/ml final) so the above is sufficient for 5 L of media. £39 is not a lot and may avoid questions when you come to publish your work.
Gary
Thanks Gary,
the reason i wanted to go for OTC kind insulin is because of easy availability and less about the price. Sigma takes around 2 weeks to a month to deliver here in India. I wanted to make a media (not available commercially) and required it at the earliest possible, hence would like to avoid the long wait for the reagent. Additionally, i just need to make 500 mL of the media at the maximum and getting the commercial one, i felt is an overkill.
Thanks Mamatha,
would you provide me with the make/supplier (as you have already tested it, i guess it would be safe)
thanks in advance...
Hi Shadi,
i guess your question was to me, rather than Gary who answered my question. (If this is the case),,,no, i did not use the human insulin for injection, but instead waited for the Sigma derived insulin. Apparently, i had to wait for a few more special reagents (other than insulin), so i had no option but to wait, hence opted to order insulin too.
you may directly ask Mamatha, who previously answered that she is using the commercial off the shelf insulin (injection grade).
regards,
Dr. Ratheesh Raman, Ph. D.
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