I need some information about corn (zea mays) cell lines. How do they grow? Are they fast growing cells? How often they need sub-culturing, etc?
Plant systems do not respond very well to in vitro liquid culturing conditions, compared to animal systems, and most commonly are in the form of protoplasts that are considered to be very stressed cells (their cell walls have been removed). In general, tissue culture is performed on semi-solid media, and it seems that developmentally, these cells behave very differently from plants (this makes sense, as the callus remains a pluripotent mass of cells, with shoots forming with appropriate hormone supplements), and show very different methylation patterns relative to tissues isolated from plants. Look at this protocol to see if it is applicable to your project: http://www.tandfonline.com/doi/full/10.4161/psb.25891#.VOUw11PF-98
Thanks Dear Johan,
I don't want to let the cell differentiate. I need them as a suspension culture with continuously sub-culturing. However, preferably the cells with the cell wall and not fast growing.
Best
I think a better word for using in plants is 'totipotency'. 'Pluripotency' is generally used for animal stem cells, which also can be induced to develop into different cell types.
Good point - totipotent is the correct term, Yuan-Yeu.
I can't think of any instances where cells will remain in suspension (I think they will readily form a callous and grow until shoot- and root-forming hormones are present, but not remain as single cells), but your question about continuous suspension culture goes past my expertise. You may find more information about continuous plant tissue culture at this website: http://aggie-horticulture.tamu.edu/tisscult/Microprop/microprop.html
There is one tobacco cultivar called BY-2 (BY: Bright Yellow), which remains in cell suspension culture for research use. Tobacco BY-2 cells are nongreen, fast growing plant cells which can multiply their numbers up to 100-fold within one week in adequate culture medium and good culture conditions. These cells have been used for different biological researches, and are compared to HeLa cells for the animals. However, I don't know if any 'BY-2'-like cell lines for the Maize.
http://en.wikipedia.org/wiki/Tobacco_BY-2_cells
1 How do they grow?
shake the callus in liquid media, more callus less liquid media, 25oC 150 rpm, sub-culturing every week. every time, pour small callus into a new flask.
2 Are they fast growing cells?
You must keep the small callus in flask, otherwise，the callus will be bigger and bigger, and never be suspension cell. The growth rate dependent on the amount of callus in flask.
3 How often they need sub-culturing,
every week.
@Dongping,
1. "You must keep the small callus in flask, otherwise, the callus will be bigger and bigger".
Did you mean 'merge' the callus under liquid medium? Because they were already in the flask. Just want to make sure. Or were you emphasizing on 'small' callus?
2. Is that a corn cell suspension culture in the attached picture?
Thanks.
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