Hi all,
At the the moment, I do co-culture experiments between T-cells and antigen presenting cells in 96-well U-bottom plate wells. These are good for co-culturing about 5x10^4 APCs per well. However, I need to scale up. Has anyone done any optimization of larger cell culture vessels that would allow for these types of co-cultures? ie. multiflasks, bottles, shaker flasks, etc. What's worked well for you in your experience?
Dear Govinda,
scalling up (as well as scalling down) does alter the manner of cell interaction.
(a) scalling up of homogeneous population (i.e. a cell-line or a cell clone) is less problematic because the neighbor cells are of the same kind; in such instances the issue is only the potential contact-inhibition when crowding occurs
(b) scalling-up in heterogenous populations, depends on the ratio of different cell types; and it can be from experimental system to experimental system different; I suggest that you first compare the trivial difference between „U“ shape and flat bottom well; 100 ul and 200 ul, If you obtain the same or similar results, then your system is robust enough to go 1 ml cultures (with two cell densities always increased by a factor of three) as well as increased volume in the same kind of 30 mm dishes (or 6 well plates); upon this you can go to large flasks (not on shakers, stationary conditions with lids not screwed tight); the volume should be such that it makes at least a 4 mm layer when horizontally incubated
(c) Scalling down; although you do not ask that, you should consider to culture smaller cell numbers (maybe even gling to Terasaki plates) to find out how close are you to limiting dilution conditions
Good luck
Ivan Lefkovits
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