BrdU analysis of mouse keratinocytes

Derek Davies daviesd2 at icrf.icnet.uk
Tue Dec 22 09:56:28 EST 1998



Hi All,

I have been doing some work with a colleague looking at proliferation in a
primary culture of neonatal mouse keratinocytes which are grown in a low
calcium medium.

Looking at simultaneous BrdU and PI analysis we see what appears to be
reduplication of DNA without division ie an 8n peak. I believe that this
has been reported to be found in terminally differentiated human epidermis
but does anyone have any experience with a mouse model? The percentage of
cells that make it to 8n is quite large (20%+). 

We thought at first that maybe the first peak was apoptotic cells
but looking at the cells using a laser scanning cytometer, the 2n, 4n and
8n peaks show single normal-looking nuclei progressively getting larger so
I think it is a real phenomenon and not a flow artefact. I suppose it
could be media-related so that is what we will be looking at next. The
thing that worries me is the large number of cells that appear polyploid.
Anyone know whether this is normal?

Thanks!!

Derek


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*  Derek Davies                       Voice: (44) 0171 269 3394            *
*  FACS Laboratory,                   FAX: (44) 0171 269 3100              *
*  Imperial Cancer Research Fund,     e_mail: derek.davies at icrf.icnet.uk   *
*  London, UK                                                              *
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