metaphase...

Jose I. de las Heras jose.delasheras at hgu.mrc.ac.uk
Fri Nov 17 15:20:36 EST 2000


Jose I. de las Heras wrote:
> 
> Penny May Avoles Kianian wrote:
> >
> > Hi
> >          I don't know if my experience can be of help to you.  We work with
> > plant root tips.  To synchronize them we either treat them with colchicine
> > for a few hours or some researchers report using cold water to synchronize.
> > Colchicine probably won't be an option since it breaks down the spindle
> > fibers and stops cell division progression.  The cold water approach might
> > be more feasible for what you are doing.  Unfortunately, I am not fimilar
> > with the technique to provide additional details.
> > Penny
> 
> The cold water treatment (0-2 degrees) works very well for plant root
> tips (and no toxic waste!). How long you treat the root tips for varies
> slightly with different species. For wheat I used to treat them for
> 20-24 hours. However, I am not sure how "happy" fibroblasts would be at
> those temperatures for long periods. Plants are much more
> temperature-tolerant, as the environment changes so much.
> 
> To synchronise divisions (again this was in plants) Hydroxyurea is often
> used. This involves a relatively short treatment which arrests growth
> (at the end of G1?), followed by removal of the HU and growth resumes...
> I can't remember what HU does to the cells, but it may be worth
> checking.
> 
> Good luck,
> 
> Jose


Also, I forgot... with the colchicine path, you'd probably be able to
get polyploid fibroblasts (I assume they are diploid now, so you could
generate tetraploids like that, providing the gene balance doesn't get
all screwy... in plants we used to double chromosome complements just
like that... ah, memories :) 

Jose


-- 
Dr. Jose I. de las Heras	Email: 	Jose.delasHeras at hgu.mrc.ac.uk
MRC Human Genetics Unit		Tel: 	(0131) 3322471
Chromosome Biology Section		ext: 	2316 (office)
Western General Hospital			2311 (lab)
Edinburgh EH4 2XU		Fax: 	(0131) 3432620	
Scotland - United Kingdom	

"Musha ring dum a doo dum a da, Whack for my daddy-o"






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