I have performed NSC culture of P0-brain and E14 spinal cord whcih never need enzyme to dissociate the neural tissue. the mechinal dissociation worked well with 1ml tip by gentle blow around 50-60 T. * A: j4 X8 H( P3 r* g5 @* ?* ~0 @. i& j0 W# L9 Y$ [
After several passage, the Neurospheres turned to be very tight then need trypsin for dissociation.