Hello All,
I have been culturing SH-SY5Y cells and have observed that after plating cells (10,000/well) in 96 well plate, they start clustering by 48 hours (big countable clusters). I am not sure if this is okay for them to cluster and proceed with my experiments such as MTT, enzymatic assays, etc. with cells in this state. If not then please give suggestions for a similar problem and how to overcome it.
Hi Himanshi Gahlot
I believe that the biggest problem causes clumping is the plate itself that some plate surfaces have a high potential tension so the cells are gonna swim and cluster in the conical bottom of the plate. If you have a reach to try different plates from different firms I highly suggest that.
On the other situation, tilt the plate in 45 degree and empty the cell suspension slowly to avoid the round-shape movement of the cell that causes a direct migration to the middle of the well. Also, avoid adding huge amount of the medium while seeding, in another words if the suggested capacity of the well is 100 ul then wet the surface of the well with a suitable volume of the medium (e.g. 25 ul), then add a 50 ul of cell suspension in a manner that prevent cell swimming. Then knock on the side that you added the suspension toward, draw plus sign 5 up-down and 5 right-left (be careful not to draw the infinity sign), after ~ 6 hours (where the cells would be set down) complete to the suggested volume.
Best....
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