Hi all,

I'm trying to isolate zebrafish embryonic stem cells after 3 hpf of zebrafish embryo's. I have a couple of questions.

- I already tried Pronase to dechorionate, but it is very difficult. I think dechorionating manually is a better way, do you agree?

- If I dechorionate manually, still the inner cell mass separates from the yolk very easily, is this a problem? Is it toxic for the cells (I don't know if the yolk contains toxic substances)? I filter through a 70 µM filter to have a pure cell culture and get rid of other debris.

- The cells do not attach to the plastic dish, will this be solved by coating the plates? Gelatin/laminin/poly-d-lysine ...?

- take a look at the pictures attached. Is this how the cells are supposed to look like?

Thank you!

Best,

Dorien