Generation of neuronal cells from pluripotent stem cells

The method for differentiating mPS cells according to claim 22, wherein the mPS cells are differentiated into cortical pyramidal neuron like cells.

The method for differentiating mPS cells according to claim 23, wherein the mPS cells are differentiated into neural progenitors of ventral forebrain identity, wherein the neural progenitors of ventral forebrain identity are positive at least for nestin and for any or all of homeobox protein GSH2, homeodomain transcription factors NKX2.1 and NKX2.2, and homeobox proteins Dlx1 and Dlx2.

The method for differentiating mPS cells according to claim 23, wherein the mPS cells are differentiated into cortical or striatal inhibitory interneuron or striatal projection neuron like cells.

The method according to claim 22 or 23, wherein the mPS cells are non-human.

The method according to claim 22 or 23, wherein the medium comprises insulin.

The method according to claim 22 or 23, wherein the medium lacks external inductive morphogens, in particular lacks components that may otherwise induce caudalisation of neural progenitors, more in particular lacks any or all of serum or plasma;

• retinoic acid (RA);
• any members of the fibroblast growth factor (FGF) family of proteins;
• and any members of the Wnt family of proteins.

The method according to claim 22 or 23, wherein the duration of the culturing step b) is between 10 days and 14 days.

The method according to claim 22 or 23, wherein the duration of the culturing step b) is at least 21 days.

The method according to claim 22, wherein the antagonist of the sonic hedgehog (SHH) signalling pathway is cyclopamine or a functional derivative thereof.

The method according to claim 23, wherein the antagonist of the Wnt signalling pathway is DKK or a functional fragment or derivative thereof.

Neural progenitors or neuron like cells or populations comprising such, obtainable or directly obtained using the method according to claim 22 or 23.

A method for differentiating mammalian pluripotent stem (mPS) cells comprising the steps of:

• a) plating undifferentiated mPS cells onto a substrate which allows adherence of cells thereto;
• and
• b) culturing the mPS cells of a) which have adhered to said substrate in a medium permissive to differentiation of the mPS cells comprising exposing the cells to an antagonist of the sonic hedgehog (SHH) signalling pathway during at least part of the culturing step.

A method for differentiating mPS cells comprising the steps of:

• a) plating undifferentiated mPS cells onto a substrate which allows adherence of cells thereto;
• and
• b) culturing the mPS cells of a) which have adhered to said substrate in a medium permissive to differentiation of the mPS cells comprising exposing the cells to an agonist of the SHH signalling pathway and to an antagonist of the Wnt signalling pathway during at least part of said culturing step.