Re: quantitation of fetal cells by flow
T. Scott Thurmond (thurmons@ehsct7.envmed.rochester.edu)
Wed, 26 Jun 1996 15:21:51 -0400
steven micko wrote:
>
> I don't know about measuring fetal RBCs in maternal blood, but I was
> surprised when we tried to run our stem cell panel on umbilical cord
> blood and found the RBCs very difficult to lyse by conventional means:
> NH4CL, R&D Systems kit, etc. Does this add any light to the subject?
> In addition, and along the lines of studying fetal/cord/maternal
> blood, I would like to know:
>
> (1) Why cord blood RBCs behave different from adult peripheral blood
> and fetal blood under the standard lysis conditions.
>
> (2) Does anybody know of a way to lyse cord RBCs and leave the WBC
> intact?
>
> Thanks
Steve,
I may have a least a partial answer for part 1 of your question. Several
studies, including my own, have shown that rbcs undergo changes in their
osmotic fragility properties in response to increased levels of estrogen. In
general, they become less fragile with increasing estrogen level. Placental
secretion of estrogen near term can be several hundred times the daily
production in the middle of normal nonpregnant monthly cycle. These
greatly increased levels, in theory, would probably change the overall
fragility profile. I would suggest experimenting with various salt
concentrations in your lysis buffer. If you would like references e-mail me
off list.
Scott
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
