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Conditioned Medium


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3 replies to this topic

#1 TomWhite

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Posted 02 November 2010 - 10:34 AM

I am planning some experiments to look at various effects of conditioned medium of cultured primary skeletal myoblasts and myotubes (after differentiating the myoblasts) and am concerned about the effects of culturing these cells under serum free conditions (required to eliminate the confounding effects of serum derived factors in the downstream assays). I am concerned that this will be a stress to the cells which will cause them to alter their normal secretome???

Any thoughts regarding this would be greatly appreciated. It seems to be an extremely common practice to culture cells in serum free medium (even for days)to generate conditioned medium but the stress/changes that this may cause to the cells never seems to be discussed.
Thanks!!!

#2 AnnaSh

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Posted 23 February 2011 - 04:34 AM

You're right. When cells are serum starved for a long time, they undergo apoptosis. You could use a protein mix known as Serum Replacement in place of classical serum. It doesn't contain growth factors, but it greatly limits loss of cell viability. I found it helpful! Bye

#3 TomWhite

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Posted 23 February 2011 - 09:09 AM

Thanks!!! I've been using MEM vitamins and non-essential amino acids which also seems to help maintain the cells.

#4 bob1

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Posted 23 February 2011 - 02:22 PM

There are a number of non-animal derived and fully defined serum replacements out there, I think Invitrogen would be a good place to start if you are interested.




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