Accelerated Induction of Cortical Neurons from hiPSCs

This protocol is intended as a guide only, for full experimental details please read the reference provided.

In Brief

Qi et al. describe an approach for the accelerated induction of early-born cortical neurons from human induced pluripotent stem cells.  At day 8 of differentiation the resulting neurons were transplanted into postnatal mouse cortex and established long distance connections. In addition, cells remaining in culture exhibit functional electrophysiological properties by day 16.

hiPSCs were plated one day prior to differentiation in conditioned medium. KSR medium was used to start differentiation the next day and was gradually replaced with N2B7 medium from day 4, reaching 100% at day 8.

A Small Molecule Cocktail for Accelerated Induction of Cortical Neurons from hiPSCs

Cocktails

Conditioned hESC MediumDifferentiation Medium Day 0-6Differentiation Medium Day 2-8
Y-27632 (Cat.No. 1254)10 μMLDN 193189 (Cat.No. 6053)10 μMPD 0325901 (Cat.No. 4192)1 μM
FGF210 ng/mlSB 431542 (Cat.No. 1614)10 μMSU 5402 (Cat.No. 3300)5 μM
  XAV 939 (Cat.No. 3748)10 μMDAPT (Cat.No. 2634)10 μM
Accelerated Induction of Cortical Neurons from hiPSCs

Reference

Qi et al. (2017) Combined small-molecule inhibition accelerates the derivation of functional cortical neurons from human pluripotent stem cells. Nat.Biotechnol. 35 154. PMID: 28112759