Transdifferentiating Fibroblasts into Neurons

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

Li et al. describe a protocol that uses a cocktail of small molecules to differentiate fibroblasts directly into neurons.

Mouse fibroblasts were cultured for 4 - 5 days. Fibroblast culture medium was then exchanged for induction medium, which resulted in cells with neuron-like morphology. After 20 days these cells were co-cultured with astrocytes in maturation medium, resulting in functional neurons.

https://resources.tocris.com/images/protocols/stemcells-image-1.jpg

Cocktails

Induction Medium Maturation Medium Survival enhancer
ISX 9 (Cat.No. 4439) 20 μM Forskolin (Cat.No. 1099) 10 μM Y-27632 (Cat.No. 1254) 2 μM
Forskolin (Cat.No. 1099) 100 μM bFGF 50 ng/ml    
CHIR 99021 (Cat.No. 4423) 20 μM BDNF 20 ng/ml    
I-BET 151 (Cat.No. 4650) 0.5 - 2.0 μM GDNF 20 ng/ml    
https://resources.tocris.com/images/protocols/stemcells-image-5.jpg?v=1

Reference

Li et al. (2015) Small-molecule-driven direct reprogramming of mouse fibroblasts into functional neurons. Cell Stem Cell 17 195. PMID: 26253201

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