I am following 2004 Nature Neuroscience paper to make neurons ( Volume 7 Number 9 Page 1003 "Differentiation of mouse embryonic stem cells into a defined neuronal lineage").
The attached picture is EB after 7 days, 4 days in ES medium and 3 days in ES + 5 uM RA. Does it look too confluent? They look very dark under scope and aggregate into big clumps.
EB too confluent?
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