RE: antigen expression after freezing

From: Nicholson, Janet (jkn1@cdc.gov)
Date: Fri Mar 05 1999 - 13:01:03 EST


If you get replies that are not posted to the board, could you post them for
general information?  To my knowledge, CD62L is the most labile antigen, as
far as stability.  I am not aware of problems with CD3, CD4, or CD8, but
then I haven't systematically looked at fluorescence intensity in cells
frozen for various periods of time.  When they are there before freezing,
they are there after.  CD38 expression is supposed to be well-preserved with
freezing (anecdotal observations from Janis Giorgi).


> -----Original Message-----
> From:	Glenda, M, Davison [SMTP:gmd@samiot.uct.ac.za]
> Sent:	Thursday, March 04, 1999 12:27 PM
> To:	Cytometry Mailing List
> Subject:	antigen expression after freezing
> 
> 
> Dear All,
> 
> I would really appreciate any advice on this question. We have been 
> freezing lymphocytes on patients post bone marrow transplantation 
> since 1995. After thawing one of the samples to perform 
> CD45Ra and RO analysis I was horrified to find that it had only 
> +/- 16% viability (using trypan blue). I set it up anyway and was 
> surprised to get good CD3 and CD45 positivity. CD4 and CD8 did not 
> give satisfactory results and it was difficult to distinguish between 
> positive and neagtive peaks. The question is: Are certain antigens 
> preserved in cell death? I dont have much experience in the 
> effects of freezing on cells and would therefore appreciate any 
> advice.   
> 
> Thanks,
> Glenda Davison
> Dept.Haematology
> Groote Schuur Hospital
> Cape Town South Africa
> e-mail:gmd@samiot.uct.ac.za



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