Hello, I work on the differentiation of stem cells into cardiomyocytes. For this I use the PSC Cardiomyocyte Differentiation Kit (ThermoFisher). After differentiation (day 14) I can't dissociate them. I use TrypLE 10X. Does anyone use this kit? How did you manage to dissociate and multiply the cells? I will be very grateful for your help.
Hi,
In my opinion, optimization of the incubation time with TrypLE 10x is necessary. Try increasing the incubation time with TrypLE 10X. Incubation at 37C is more effective. Incubate with TrypLE 10x for 5-15 minutes at 37C. Observe cell morphology during incubation.
Stop incubations when single cells will be visible. Add a ROCK inhibitor.
Use Dulbecco's Phosphate Buffered Saline (DPBS) without calcium and magnesium.
Justyna Augustyniak
Dear Irina,
Which kind of ECM proteins did you use for the pre-coated cell culture plate?
Did you use Matrigel? Tell me the concentration.
10X TrypLE is too high for cells, and treatment with this high concentration definitely would kill the cells!
I use 0.5X TrypLE for 3-5 minutes to cell dissociation.
- Badges
- Science method