Direct Neuronal Reprogramming of Mouse Astrocytes into Neurons
Transcript
Prepare neuronal differentiation medium by adding one milliliter B27 supplement to 49 milliliters of basic culture medium. After 24 to 48 hours, depending on whether cells have been transduced or transfected, replace astrocyte medium with one milliliter neuronal differentiation medium per well. Culture the cells at 37 degrees Celsius with 9% carbon dioxide.
To increase the reprogramming efficiency, supplement the neuronal differentiation medium with Forskolin and Dorsomorphin to a final concentration of 30 micromolar and one micromolar respectively when replacing the astrocyte medium with the differentiation medium. When opting to treat cells with Forskolin and Dorsomorphin, provide a second dose of Dorsomorphin two days after the initial treatment. Add this second treatment directly to the culture medium.
