fine arabidopsis suspensions

Nobody nobody at hgmp.mrc.ac.uk
Mon Sep 4 16:00:24 EST 2000


Arabidopsis suspension cultures:  How to get a fine suspension?

We have been culturing arabidopsis suspension cells but have a problem
with friability in liquid culture.  The callus produced from seedlings
on plates containing MS/5mg/L 2, 4-D/3% sucrose grows well and is
friable.  When this is transferred to liquid culture with the same
nutrients and hormones the cultures produced are very clumpy.  The
clumps of cells seem to grow (slowly) but don=92t break up easily.  There
are some finer cells, presumably sheading from the large clumps, these
are predominantly single cells and do not appear to grow when selected
(by seiving) and used to inoculate new flasks.  We have also tried
MS/0.5mg/L 2, 4-D/0.05mg/L kinetin/3% sucrose and Gambourgs 1mg/L 2,
4-D. 0/04mg/L kinetin/2% sucrose in liquid culture but got the same
outcome.

Does anyone have advice on how to generate a fine culture?

If you have success please advise on:

1) Medium
2) Sub-culture procedure
3) Sub-culture period
4) Type of flasks/bottles
5) Shaker speed
6) Sieving?
7) How long does it take to generate a fine culture?

Thanks very much,

Humphrey Foote


************************************
Humphrey Foote, PhD (h.foote at genesis.co.nz)
Genesis R&D Corp Ltd (www.genesis.co.nz)
PO Box 50
Auckland
New Zealand
Tel: 64-09-373-5600
=46ax: 64-09-373-2189
************************************


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