Growing caco-2 cells, but can not get cell count due to aggregation of cells (bunch of 10-20 cells). Already tried pipetting and vortex but still there are cell agrregate making it difficult to count then with the help of haemocytometer.
I have a few hints:
1. Wash your cells throughly with PBS magnesium and calcium free prior to trypsinization.
2. Always passage CaCO-2 when they are 70% confluent. This is specially troublesome for CaCO-2 cells because they change cell-surface morphology upon reaching confluence.
3. Be careful not to over-incubate your cells with trypsin - aggregates are common in longer incubations, also impacting your cell viability. I normally use 0.25% trypsin for 3 min.
Reason for washing them with PBS before is that you have to remove the rest of the medium since there are substances which are inhibitin* the trypsin. If this does not help (if you had a good teacher you should have known that) do not Pipette them up and down but use a syringe with a bigger tip and do it max 2 times. This worked for me and my HepG2 cells, which are very sticky. Just remember: keep them in cultUre for at least 2 days after to make sure they are not stressed anymore. Otherwise they might react strange. Good luck
