responses to Arabidopsis cell culture

Jodi Swidzinski jodi.swidzinski at plant-sciences.oxford.ac.uk
Mon Jan 24 14:12:13 EST 2000


For those of you who wanted to see the responses to the cell culture
"clumping" query, these are some of the responses I received. Thank you to
everyone who helped me out.
JS


If you filter your cells through an 80 micron mesh and then "scoop" some
cells (not the clumps) with a spatula into a new flask with medium, you
should be able to get rid of the clumps. The media I used can be found here:
http://www.msu.edu/~delgadoi/102.html
     Good luck,
     Ivan



If your concern is simply for ease of examination under the
microscope, it may be worth transferring your cells for examination
into 0.1M EDTA (pH 8.0)  for an hour or two before you wish to look at them.
The EDTA chelates the calcium from the cell walls, and loosens the
intercellular connections.

If the clumping is preventing you from getting accurate cell counts
for estimating culture density, this may not be of much help.

Best wishes
Liz Kinsman



e.kinsman at roehampton.ac.uk

Liz Kinsman
Senior Lecturer (Plant Biology)
School of Life Sciences
Whitelands College
University of Surrey Roehampton
London SW15 3SN

0208 392 3534


Diluting the cells in water and vigorously vortexing them can
partially disassociate clumps.  Additionally, pellet the cells and
resuspend them well in a sterile 0.1% agar solution.  You might also
trying to growing the cultures in media containing 0.1% agar.  This
concentration of agar is a liquid, not a solid.




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