I am culturing patient-specific iPSC-derived astrocytes in a monolayer. So far I have cultured them in wells of a 6-well plate and they have been happy. I have a very small number of cells and would like to maximise what I can test, therefore use the smallest coverslip. I heard the smaller the well, the unhappier the cells.
Any advice or suggestions would be greatly appreciated!
If you are culturing the same number of astrocytes with the same amount of medium on a smaller area, as you did in the 6-wells, the only difference is that it will be faster to reach confluency. Once it reached confluency (depending on the seeding concentration, usually after 2 days), recommended that you maintained the cells by changing the medium or fixed it immediately for your other tests. Good lucks with your research work!
What makes the difference is the surface/volume ratio of your culture liquid media. As higher this ratio the better is gas exchange. There is a lower limit as well off cause for several reasons. So I guess you have to try, whether they are happy in smaller wells at the same surface/volume ratio.
I've cultured primary mouse astrocytes in 24-well plates without problems. Just make sure your seeding density is not too high and "downsize" proportionally.
I agree with what everyone has stated. I have used both 12 well and 24 well plates and the only difference if seeding them at the same density is the time they will reach confluency. Change the medium every two day and you should not have a problem. I suggest you base your plating strategy on what experiments you plan to do.
There should not be any problem if you scale down cell number and volume as suggested. You mention using coverslips for the smaller cultures; you do this as well when culturing in 6 wells?
Remember also that your output (endpoint measurement) will be reduced accordingly when you switch plate sizes, i.e. you'll have less protein, less cytokine release and so on (but not a problem if your are doing IF/IHC).
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