Weekend free protocols for iPS maintenance

hiPSC Culture Discussion
1

Can I passage cells on a Thursday or a Friday and do a double feed on a Saturday so that I can skip a feed on a Sunday?

2

Hi hperera! I routinely passage my cells on Thursday at a density where I can double feed on Friday and passage again on Monday (no feeding required on Saturday OR Sunday- hooray!).

For cell lines that do not require ROCK inhibitor when passaging (such as the parental WTC11s), I have also had success passaging on Friday directly into 6-well plates containing 4mL of medium per well and incubating until Monday.

Note: I use mTeSR Plus medium

1 Like
3

Hi en2b!

Thank you for the information. What is the seeding density that you use for 6well plates? when passaging on Friday?

4

Hi @hperera,

We typically seed 150K cells for 3 days, 75K for 4 days in 6 well plates. We find that growth is more dense at perimeter of the well and uneven at the center.

Thanks,
Janani