Someone has experience in cell culture of skin fibroblasts of domestic cats ? Upon reaching 50 % confluence the cells began to detach from the edge of the culture plate. We have prepared new medium (DMEM - F12 + 10 % FBS) and pH adjusted , cleaned and checked the incubator temperature , We unfreezed new cells and it is happening the same thing.. Anyone know if the cells of domestic cats require some kind of special culture ? Anyone know what might be going ?
I agree coating of the plates will help with adhesion, we tend to use either 50ug/ml collagen in 0.02M acetic acid or 0.1% gelatin as above.
Losing cells at the edge of the wells may also be a consequence of harsh washing/media change. Make sure the PBS/media is run gently down the well sides if this is the case.
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