Plant cell culture- How do prevent differentiation of callus cells - (Feb/26/2007 )

Hi all!

Im trying to grow callus of a cactus plant, I'm trying to get friable or soft cells for cell suspension culture but I'm facing the problem of callus differentiation. I need my cells to be remain as callus cells.

PLEASE HELP!

Leeann

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Hi!
Is it a new protocol that you are testing? Have you ever noticed the same thing in other cultures???
I think that your cells behavior depends most on the Plant Growth Regulators you choose.
They are the crucial factor to force your cells to be in your desired status.
Anyway, some cultures are very difficult to influence and you can't avoid completely a differentiation phase.
Maybe you have to think about introducing different culture steps to build up a cycle.
Anyway, the most important things to keep in mind are -in my opinion- the optimum of PGRs combination and the time you decide to leave your cultures on the same media.
Unfortunately the setting of culture protocols is always quite long and labor-intensive...

E.g.:
I can't eliminate the differentiation step from my morphogenic cultures of callus, otherwise they will loose their embyogenic ability.
So I alternate two media:
two months: the first substrate contains more cytokinines than auxines so it induces callus proliferation (= I have lots of undifferentiated cells!!!)
two months: the second substrate contains more auxines than cytokinins, so it induces embryos formation and differentiation.

You can also regulate the other factors that influence callus differentiation, as - for example - light or the type of medium.

Don't hesitate to ask for further explanations, in particular if I was not clear enough (and it's probable.....)
Hope it will be of some help!
Good luck!
Ila

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Hi Ila

Thank you so much for replying, you see the problem is i dont want my callus cells to differentiate, I want them to remain callus cells, another problem is the contamination within the callus cells, i have tried so many antibiotics, nothing has worked, they just kill my callus cells, and my callus cells of a cactus plant just wouldnt grow in liquid medium. DAME THIS IS SO HARD!!!!!!

Leeann

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