Accelerated Induction of Cortical Neurons from hiPSCs

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

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 term 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.

https://resources.tocris.com/images/protocols/neural-stem-cells-1.jpg

Cocktails

Conditioned hESC Medium Differentiation Medium Day 0-6 Differentiation Medium Day 2-8
Y-27632 (Cat.No. 1254) 10 μM LDN 193189 (Cat.No. 6053) 10 μM PD 0325901 (Cat.No. 4192) 1 μM
FGF2 10 ng/ml SB 431542 (Cat.No. 1614) 10 μM SU 5402 (Cat.No. 3300) 5 μM
    XAV 939 (Cat.No. 3748) 10 μM DAPT (Cat.No. 2634) 10 μM
https://resources.tocris.com/images/protocols/neural-stem-cells-2.jpg

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

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