Table 3.

Brief overview of neuronal differentiation protocol from Shi et al. and adaptations made in this study.

Protocol	Neural Maintenance Medium	Neural Induction Medium	Supplements	Notable Differences
Shi et al. 2012	N2/B27 medium. N2 medium: Basal medium with 1% N-2 supplement, 1% 200mM L-alanyl-L-glutamine dipeptide in 0.85% NaCl solution, 1% pen/strep, 5ug/mL insulin, 1mM ;-glutamine, 100μM non-essential amino acids, 100μM 2-mercaptoethanol. B27 medium: DMEM/F12 supplemented with 2% B-27 supplement, 200mM L-glutamine, 1% pen/strep.	N2/B27 medium supplemented with 10μM SB431542 and 100ng/mL noggin OR 10μM SB431542 and 1μM dorsomorphin.	20ng/mL FGF2 upon appearance of rosettes	Use of Insulin, NEAA, 2-mercaptoethanol
This protocol	N2/B27 medium. N2 medium: Basal medium with 1% N-2 supplement, 1% 200mM L-alanyl-L-glutamine dipeptide in 0.85% NaCl solution, 1% pen/strep. B27 medium: DMEM/F12 supplemented with 2% B-27 supplement, 1% 200mM L-alanyl-L-glutamine dipeptide in 0.85% NaCl solution, 1% pen/strep.	N2/B27 medium supplemented with 10μM SB431542, 1μM dorsomorphin, 100nM LDN193189.		Use of 200mM L-alanyl-L-glutamine dipeptide in 0.85% NaCl solution in B27 medium, use of 100nM LDN193189 in Neural Induction Medium