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Culture media for BGMK-hDAF

BGMK

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#1 Sanedor

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Posted 14 May 2012 - 03:41 PM

Hello,

New to the forum but not to the lab!
Does anyone here have experience culturing BGMK-hDAF cells? Some online research had turned up multiple different methods as to what sort of media to use however I was wondering if anyone had some testimonials as to what worked for them?

I am hesitating going either with MEM + (5%-10%)FBS + antibiotics or 50/50 MEM/L-15 + (5%-10%)FBS + antibiotics. We have a CO2 tank so L-15 would not be a necessity but published have stated this media in their methods while others have gone strictly with MEM.
Our cells will be coming in frozen (not sure if DMSO or glycerol) but the supplier was of no help as they stated that they obtain their cells strictly from the patent office???

Any input would be greatly appreciated! Many thanks in advance!

#2 Sanedor

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Posted 15 May 2012 - 02:50 PM

Well drat, I was hoping someone would have had some experience with these. Ok, well I guess my next question is does anyone have any advice, insight or thoughts that I failed to mention above?

#3 bob1

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Posted 15 May 2012 - 03:19 PM

I was going to suggest asking the supplier - who is the supplier anyway? Often these sorts of things will come from the ATCC (atcc.org) or ECACC (hpacultures.org.uk), which are the definitive sources for cell lines. Their website(s) are probably the best place to start.

Ideally the cells will also come with some information which will allow you to make an informed choice.

#4 Sanedor

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Posted 15 May 2012 - 07:22 PM

Hi bob1,
Thank you for your suggestions.

I did contact the supplier (name escapes me right now but will post tomorrow) and they claimed that they only handled the transactions. The spokesperson stated that they obtained the cells straight from the patent office and that the cells came with no instructions or any sort of information. I proceeded to contact Viromed as they seem to be the origin of the line however without an account, they of course would not divulge any information. Our lab went with BGMK-hdaf cells rather than just BGMK based on the increased sensitivity to enteroviruses.
The cells arrived today and I had to go out on a limb since I could not locate any straight forward information (I even read through the 32 page patent...). The criovial came only with a date and passage number. No cell count or anything of valuable information other than lots of disclaimers.
I proceeded to thaw and spin the cells down and perform a count and seeded them in 50/50 MEM/L-15 + (5%-10%)FBS + antibiotics.

My hopes are that since the literature displays so many different methods, the cells will not be too picky.

I failed to mention that we had prior frozen stocks in our lab however all of the vials I have tried produce nothing more than floating cells, viable but floating when in fact they should be adherent.

Thanks for reading




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