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Thymus macrophage isolation

Christopher Hatton (remove_this) "CDH8" at le.ac.uk
Tue Sep 22 05:08:18 EST 1998


Hi Youngkyun Lee,

I isolate mouse thymocytes. but these papers also isolate all the
stromal cells aswell and from which you can isolate the thymic nurse
cells (thymic "marcophages")

Nel Moore et al (1993) Analysis of Cytokine gene expression in
sub-populations of freshly isolated thymocytes and thymic stromal
cells....  European Journal of Immunology. 23: 922-927

Robert Wilkinson et al (1995) Positive selection of thymocytes involves
sustained interactions with the thymic microenvironment" "Journal of
Immunology. 155: 5234-5240

Both of these are from the group of John Owen and Eric Jenkinson in
Birmingham (UK) they have done a lot of work in this area using mice. 
However the following 2 papers mention Rat thymocytes, and with the use
of relavent antibodies you could isolate the thymic nurse cells either
by FLOW CYTOMETRY OR immunomagnetic cell selection.

Sek Chow et al (1997) Susceptibility of different subsets of immature
thymocytes to apoptosis. FEBS letters 408 pp141-146  (also Cohen et al
1993-J. Of Immunology 151 p566-574)

Prapapas et al (1993) Enrichment and characterisation of
thymus-repopulating  cells in stroma-dependent cultures of rat bone
marrow. Journal of Cell Science 104 p1039-1048

Good luck, and just to say, Macrophages per se aren't normally resident
in the normal thymus although a macrophage like cell is...the "thymic
nurse cell"  You may need a couple of young 4-5 week old
***unstressed*** rats for 1 million cells.

hope that helps

Christopher Hatton
PhD Student
Centre for Mechanisms of Human Toxicity
Leicester
(UK)

overheat wrote:
> 
> I'm a graduate student at Seoul National University, department of
> chemistry.
> I'm studying an enzyme possibly involved in apoptosis in rat thymus.
> I suppose that enzyme activity resides in thymus macrophage, so I want
> to isolate macrophage from thymus.  If anybody have a protocol for the
> isolation method (I need at least million cells per assay),  please t



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