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Data has shown direct transdifferentiation between different germ layers when transfected with a cocktail of defined transcription factor genes1,3. A supplement of this small molecule cocktail in cell culture media simplifies the process and increases the productivity of neural and neuronal cell generation.
Repro-nLSC is a cocktail of three small molecule inhibitors. It includes LDN193189, SB431542 and CHIR99021. Using the combination of synergistic SMAD inhibition through BMP and TGF-beta pathways together with specific GSK-3beta inhibition, a recent study has presented that postnatal human fibroblasts (HPF), transduced with Ascl1 and Ngn2, efficiently converted into functional neuron-like cells with yields up to >200% and neuronal purities up to >80%. In addition to the conversion of mesenchymal cells to induced neurons (iNs), this method has also been tested in the conversion of other somatic cell types (eg. Cord blood-derived stem cells) to iNs3.
Technical information:
| Cocktail Formulation: 1000xDMSO stock solution | 0.5 mM of LDN193189 (BMP inhibitor), 10 mM of SB431542 (TGF-ß1 inhibitor), and 2 mM of CHIR99021 (GSK3ß inhibitor) | |
| Purity: | >98% (for each compound) |
Shipping Condition: The product is shipped in a glass vial at ambient temperature.
Storage: Store DMSO solution at -20oC.
Usage: Dilute Repro-nLSC stock solution (1000x) at 1:1000 into your reprogramming media to initiate neuronal conversion.
Reference:
| 1. | Vierbuchen T, et al.Direct conversion of fibroblasts to functional neurons by defined factors. Nature. 2010. 463(7284):1035-41. |
| 2. | Chambers SM, et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009 27(3):275-80. |
| 3. | Ladewig J., et al. Small molecules enable highly efficient neuronal conversion of human fibroblasts. Nat Methods. 2012. 9(6):575-8. |