RE: Cord blood
Richard Moldwin (rmoldwin@midway.uchicago.edu)
Fri, 1 Aug 1997 08:01:51 -0500
On Thursday, July 31, 1997 12:11 AM, Dr William Smith
[SMTP:williams@ichr.uwa.edu.au] wrote:
>
> Hi Flow people
> I'm working on cord blood phenotyping, interested in rare leukocyte
subsets.
> As those who work on these preps will know, there is a problem with RBC
and
> erythroid contamination, variable but up to 80% of ficoll separated PBMC.
_____________________________________
> Dr William Smith
>
> Institute for Child Health Research
> (Company Limited by Guarantee ACN 009 278 755)
> Subiaco, Western Australia, 6008
> PO Box 855 West Perth WA 6872
>
> Ph 61 8 9340 8792/8388, Fax 61 8 9388 3414
> email williams@ichr.uwa.edu.au
> ______________________________________________
>
For cord blood banking, we use two approaches to RBC removal.
For red cell agglutination and removal, we with hetastarch (Hespan) to
agglutinate red cells. See Pablo Rubinstein's 1995 PNAS paper (vol 92, p
10119) for details.
For counting cells, we use "Zapoglobin" (saponin-based, I believe) to lyse
most of the contaminating red cells
I hope this helps
--Rich Moldwin
^~^~^~^~^~^~^~^~^~^~^~^~^~
Richard Moldwin, M.D., Ph.D.
Section of Pediatric Hematology/Oncology
University of Chicago Hospitals
Wyler Room C408, MC 4060
5841 S. Maryland Ave.
Chicago, IL 60637
Tel: (773) 702-6808
Fax:(773) 702-9881
E-mail: rmoldwin@midway.uchicago.edu
WWW: http://rmoldwin.bsd.uchicago.edu