How do you test for pluripotency in edited hiPSCs?

[crisprnavigator25]

Hey all, I'm working with hiPSCs and just did some gene editing. I need to check if the cells are still pluripotent afterward. What methods or tests do you use to check pluripotency in hiPSCs after editing? Any advice or protocols you can share?

 

[nucLe01]

What we typically do first is morphology assessment by looking for compact, tightly packed colonies.
Then, we go immunostaining or flow cytometry for markers like OCT4, SOX2, NANOG, SSEA-4, and TRA-1-60.
You can also confirm with qRT-PCR.
Then finally, we do the gold standard for functional pluripotency, the in vitro embryoid body differentiation to check for all three germ layers.
We've proven that this is a solid alternative to teratoma assays.