Dear Kiana,

It really depends on the type of culture vessel you are using. With rectangular shaped T-flasks, moving the culture vessel in a circular motion for 10-15 times would normally disperse the cells quite well due to the straight walls of the T-flasks connected by sharp edges. As a good practice, however, you still need to check the dispersion of your cells under a microscope especially before you place them within the incubator.

With culture vessels which are circular in shape (e.g. 100 mm dishes, 24-well plates), there are no straight sides, thus, moving the vessel in a circular motion would tend to let cells aggregate at the smooth circular edges. To overcome this, what I normally do is to move the vessel in a 'plus' sign (+) motion for 10-15 times to get a good cell dispersion. You can observe improper mixing of the cells by the presence of cell clumps (although it might vary between cell types) even before ~80% confluency after incubation or the presence of a 'halo' surrounding a clump of cells in the middle of the culture vessel. This has a lot to do with the distribution of liquid and its contents in different shaped culture vessels.

The safe practice here is to always make sure to check on your cell dispersion before you place them in the incubator so you don't affect any treatments you are going to perform on the cells after a certain time-point.

Good luck in your experiments!

Dear kiana,

It happens to me every time I  don't mix the plates verry well so the cells wil be gathered on the edges

So you should mix verry well your plates after the seeding of your cells

Dear Kiana,

It really depends on the type of culture vessel you are using. With rectangular shaped T-flasks, moving the culture vessel in a circular motion for 10-15 times would normally disperse the cells quite well due to the straight walls of the T-flasks connected by sharp edges. As a good practice, however, you still need to check the dispersion of your cells under a microscope especially before you place them within the incubator.

With culture vessels which are circular in shape (e.g. 100 mm dishes, 24-well plates), there are no straight sides, thus, moving the vessel in a circular motion would tend to let cells aggregate at the smooth circular edges. To overcome this, what I normally do is to move the vessel in a 'plus' sign (+) motion for 10-15 times to get a good cell dispersion. You can observe improper mixing of the cells by the presence of cell clumps (although it might vary between cell types) even before ~80% confluency after incubation or the presence of a 'halo' surrounding a clump of cells in the middle of the culture vessel. This has a lot to do with the distribution of liquid and its contents in different shaped culture vessels.

The safe practice here is to always make sure to check on your cell dispersion before you place them in the incubator so you don't affect any treatments you are going to perform on the cells after a certain time-point.

Good luck in your experiments!

Uniform dispersion of cells while cell seeding can prevent the gathering of cells in the edges of the culture plates.

Try to shake the flask or dish before incubation as it will distribute your cell uniformly in the specific dish. I agree with @Mamatha M.Pillai

Cells in the culture flask after trypsinized become non adherent. But what when adherent cells grown at the edges more then the centre of the culture flask. Why cells in the growing phase , grows more at the periphery rather than centre