keratinocyte culture media - (Nov/02/2005 )

Hi,

I want to culture canine keratinocytes but I am having a lot of trouble ordering cholera toxin. Does anyone know of any alternative I can use?

Thanks.

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cholera toxin?

forgive my ignorance, but what does that have to do with keratinocyte medium?

we grow human keratinocytes in KGM, can you not use some similar defined medium for canine as well, with the appropriate growth factors?

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Hi,

I am going to use Williams Medium E but in all the methods I have read both epidermal growth factor and cholera toxin must be added. I am going to try culturing them in the absence of cholera toxin but I don't know if it will work. My supervisor seems to think that it is quite important to add cholera toxin but if i have to wait 12 wks it will put me behind in my work.

I am new to all of this having just started my PhD and so I thought someone on here might be able to advise me.

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Hi ,
I also use KBM-2 media for NHEK. I have never heard of cholera toxin for normal growth. You can find out more about toxin(i.e. if any specific compund from toxic is necessary, so you can order only that perticular compound).

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I don't have experience with your cell line, but I grow MCF10A breast epithelial cells and some groups add cholera toxin to the media. I emailed ATCC and asked the purpose and the reply was that it helps the cells grow better and prolong the time they can be kept in culture due to the increase in cAMP. I don't use the cholera toxin and my cells seem fine. I think cholera toxin was something added to primary cells when someone wanted to keep them going in culture for a long time. Seems kind of weird to me. . . but I am not an expert on cholera toxin

good luck!

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Hi,

Thanks for all your replies. Thoose people who don't use cholera toxin what growth factors are you adding?

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Hi,

As I mentioned earlier I am using KBM media from Cambrex. It comes with all growth factors in seperate bullet kit. In general, it contens Bovine pitutary extract, hEGF, insuline(bovine), hydrocortisone, GA1000(Gentamicin, Amphotericin-, Epinephrine and Transferin

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for the MCF-10A cells I add the other recommended growth factors/hormones. so for my cells that is EGF (20ng/ml final), hydrocortisone (0.5ug/ml final) and insulin (10ug/ml final).

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Thanks Sarah thats really helpful the papers I have read on human keratinocytes seem to omit cholera toxin so I am going to try and do that.

Also, thanks everyone else you have all been very helpful! :

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