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Adipose Conditioned Medium and Storage of Adipose Tissue


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#1 NazGoole

NazGoole

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Posted 20 December 2013 - 11:23 AM

Hey everyone,

 

I have to do some assays with ACM and Keratinocytes and Adipose Tissue and Keratinocytes in 6-Well plates. As the fat tissue comes directly from the operation room, and I do not always know in advance when such a surgery is scheduled, I can't time my keratinocyte seeding so that i can make immediate use of the fat. For my keratinocyte culture I use Serum Free Keratinocyte Medium. Should I use it for the ACM preparation too or should I rather take DMEM or something else? Because I think DMEM, etc contains too much calcium.
I have read that I can store the ACM at -80C for nearly as long as I want to, but what about the thawing? Can I just defreeze it at room temperature, or is there a trick?
 

Another question is, how can I store the adipose tissue itself over longer periods? (A few days, until I have enough keratinocytes to be seeded, resp. until the ones I have seeded are attached)
We usually store it in a refrigerator (4°C I think) but my supervisor says that it should be used within 24 hours at longest.
As I sometimes have to wait longer then 24 hours before I am ready to use it, I was wondering if I could just put the adipose tissue into a medium and leave it in the incubator. I already kept fat tissue in keratinocyte medium for about one week (together with keratinocytes for the assay), and it looked quite healthy afterwards, but I just don't know if this alters any properties of it (reduces its metabolism, reduced release of cytokines, adipokines, growth factors, etc.).

Thank you all, Merry Christmas and a happy new year!
NazGoole


Edited by NazGoole, 20 December 2013 - 12:31 PM.





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