Re: HSC TESTING AFTER CRYOPRESERVATION

From: Ty Creelan (creelant@ucs.orst.edu)
Date: Tue Aug 08 2000 - 19:10:53 EST


My understanding of cryopreservation and cell marker
staining (CD40, CD34,&c) is that the cells must be stained prior to
freezing; otherwise, many or most of the surface molecules
are lost and their binding abilities are compromised. I guess this depends
upon what you are looking at, but I think it is best to stain the
cells first and then freeze them. Also, the comparison between samples
stained simultaneously is more accurate than that between samples stained
at different times simply because small technical variations (in the
quantity of staining reagent used, incubation time, temperature, &c) can
lead to fairly significant differences in how the cells take up the stain.
Hope this helps.

___________________________
Tyler F. Creelan
OSU Dept. Microbiology
Nash 552 737-5094

On Tue, 8 Aug 2000, Liza McGuire wrote:

> Good-day to you flowers,
>
> I am searching for guidelines and/or standards that involve testing peripheral blood
stem cell collections post cryopreservation. At this point our laboratory is testing
for viability, sterility, CD34 enumeration and CFU at the point of collection (fresh
sample) as well as at the point of infusion (post thaw). Is it necessary to do all of
the testing post cryopreservation? Naturally, the results will not correlate with pre
cryopreservation. Furthermore, they are time consuming and the decision for the stem
cell infusion dose has already been made. I would consider any post cryopreservation
data points to be useful for future clinical research studies. What do you think?
> I have checked into FAHCT guidelines and if I understood correctly the decision is
left up to the laboratory director.
> I am trying to get a feel of what the "norm" might be. What is the consensus? I
appreciate and I will be grateful for any input that you may have.
> Thanks a bunch in advance,
> Liza McGuire
> Bone Marrow Processing Lab
> Thomas Jefferson University Hospital
> 130 S 9th Street, Rm. 400 Edison
> Philadelphia, PA 19107-5233
> (215)955-9646
>
>



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