Hi , I work on mk45 cell line (one of the gastric cancer cell lines)
I want to make them single after trypsinization that can form spheroid bodies, But most of them tend to form aggregates.
Should I use any dissociation buffer to make them single?
Try to trypsin after washing the cells with 1X PBS and for a longer duration (15 min). Add media and collect cells in a 15 ml tube and use 10 ml pipette to continuously forward-backward pipetting for 15-20 times. Centrifuge at 1200 rpm for 3 min at room temp. Add desired amount of media and use 10 ml pipette again for 15-20 times pipetting. Count the cells and do serial dilutions to get single cells per well of 96 well plate.
Good luck!!
Thank you for the answer.
Do you mean that I wait for 15 min after adding trypsin? And how much trypsin is required to add into my cells with 90% confluency?
- Badges
- Science method