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Freezing PBMCs that had been thawed and cultured


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#1 neuropath

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Posted 30 January 2015 - 10:21 PM

Hi All,

 

I am going to do some ELISpot assays and I have obtained cryopreserved PBMCs from a commercial source.

 

However, the number of cells in each vial is very high and much more than what I would need for each set of experiment. 

 

Rather than wasting the extra PBMCs, I am wondering if it is possible to cryopreserve them again after a short period of culture. Would it affect the immune functions of the different cell subsets?

 

I would really appreciate some advice on this.

 

Thanks!

 

~neuropath~



#2 Trof

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Posted 31 January 2015 - 03:17 AM

I think the cells could be re refrozen (at least I think I saw colleagues do that) but I would say not after a cultivation, MCs are primary cells, one you thaw them they start to differentiate and die, not like immortalized cells. The viability of the refrozen cells may be lower though.

If you looking on the surface markers of the mononuclear cells then especially cultivation could change it as the cells slowly differentiate.

 

I would probably ask the company, if they have a protocol for spliting the vial or immediate refrozing of what other option the have. They may tell you that you can refroze them fine or.. I don't know, depends on how they were frozen in the first place.
 


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