I have converted iPSC to cardiomyocyte and I would like to make functional assays to cardiomyocyte so I need to passage them. If I used accutase for this passage, does it cause any damage to cells or does it work to deattach the cells?
It should work very well to detach your cells. The advantage of accutase over the traditional trypsin treatment is exactly that it is more gentle and less damaging to cells.
Here a protocol for your reference: https://www.promocell.com/f/2018/01/C-41310.pdf
Many thanks Alexander Trampe . How long would you suggest me to wait cells in accutase at 37C? After passaging I will continue to grow them in RPMI 1640/B27.
I would suggest starting with 5 min incubation at 37 °C - usually, that is sufficient for cardiac myocytes.
Sorry, no ideas from me - but I hope you can find some hints here in the literature:
Article Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes: ...
Definitely YES
The passaging solution(like Accutase) is much better than Trypsin for cell dissociation and keeps your cells alive
Base on your cell line the incubation time is a bit different but I recommend you 7-8 min at 37C (if you keep less your cells not deattach completely)
Remember don't pipet cells a lot
If you keep cells in optimize time the cells easy to deattach with 3-4 pipetting
please mix well your Accutase solution before use
Hello,
RPMI 1640 medium (11875093, ThermoFisher Scientific) supplemented with B27 (minus insulin) (A1895601, ThermoFisher Scientific) and 5 µM CHIR (72054, STEMCELL Technologies). Forty-eight hours later, media was changed to RPMI 1640 with B27 supplemented with 4 µM IWP2 (72124, STEMCELL Technologies).
It is not possible to collect and passage these cells.
Good Luck
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