We are trying to transition our iPSCs to a serum free media, how do you recommend weaning and transitioning?

Answer

I am assuming you are starting with some fibroblast growth medium like DMEM supplemented with ESC-qualified FBS. If you are using GIBCO Essential 8 medium as your SFM you can transition directly. Essential 8 is designed for feeder free growth. Here is a reference https://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols/generation-human-induced-pluripotent-stem-cells-fibroblasts.html

If not, There are two ways to do this. Try a sequential adaptation. Assume you are using old media A and want to transition to new media B. Start in media A and know something about the kinetics of growth. Transition to 80%A:20%B grow several passages then move to 60%A:40%B then 40%A:60%B and so on until your cells are in 100new medium B. The other method involves removing 50% of the old media and supplementing with 50% the new media. If you are removing serum you will need to coat the dishes with some attachment matrix like geltrex or vitronectin if you want attachment.

Pin It on Pinterest