Dear Kay,
PBMC numbers are quite variable from blood sample to blood sample. Recovery of
leukocytes from a buffy coat (of a 450ml blood bag) is generally between 200-800
million cells. Roughly 30% are lymphocytes, about half of these T cells. In
healthy persons, about 50% of these are CD4+. Naive CD45RA+ finally anywhere
between 5-25%, i.e. you should start with the buffy coat of at least one regular
blood donation (450 ml). You can get these at no or very little charge from the
Red Cross blood donation centers. I can send you a protocol for purification if
you wish (it is in German, though).
Oki
kay schrieb:
> I am trying to sort out naive T cells (CD45RA+). This is a subset of
> PBMC,of CD3+T cells of CD3+CD4+ T cells. Last time, I did not get
> enough cells from 17ml of blood for my stimulation experiment ( I need
> at least 2,500,000. I need to know the number of lymphocytes in say a
> ml of blood and the percentage of each subset I should expect out of
> that number. That way I can save myself some BLOOD, sweat and tears.
--
___________________________________
Dr. Octavian Schatz
Labor Infektiologie
Med. Poliklinik
Pettenkoferstr. 8a
D-80336 München
Tel. +49-89-5160-4929
Fax +49-89-5160-3593
e-mail: schatz at pk-i.med.uni-muenchen.de
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