My neural induction media had 3 uM CHIR, 2 uM SB, 2 uM DMH1 all resuspended in 99.9% DMSO (had some issue in making 1 mM stock; powder was not measurable for 1mg), 0.5X N2 supplement, 0.5X B27 supplement, 0.1 uM ascorbic acid (made in PBS), 1% glutaMAX, DMEM F12 + Neurobasal medium (1:1).

Here are my doubts in detail.

1. Is this how the cells are supposed to look or they look stressed?

2. DMSO has caused any stress in culture?

3. Do the cells look like they are on the path of differentiation?

Please suggest me some tips for the same.