Can PSC Neural Induction Medium be used to differentiate hPSCs into neurospheres or neurons?
PSC Neural Induction Medium is a medium to convert hPSCs into NSCs. At day 7 of neural induction, dissociate P0 NSCs with Accutase. The following recommendations can be use for differentiation of neurons or neurospheres:
For differentiation of neurons: Use PSC Neural Induction Medium to convert hPSCs into NSCs. At day 7 of neural induction, dissociate P0 NSCs with Accutase. If dissociated NSCs are plated on a Geltrex-coated culture vessel in neural expansion medium, NSCs will grow as a mono-layer. For NSC expansion, it is necessary to treat NSCs with 5 µM ROCK inhibitor Y27632 at the time of plating to prevent cell death if NSCs are under P4.
For differentiation of neurospheres: Use PSC Neural Induction Medium to convert hPSCs into NSCs. At day 7 of neural induction, dissociate P0 NSCs with Accutase. If dissociated NSCs are plated into a non-coated flask in neural expansion medium, dissociated NSCs will re-aggregate into small spheres. NSCs in each sphere will proliferate and form big neurospheres. We have not expanded NSCs in neurosphere format in-house. You can test expanding NSCs in neurospheres with and without ROCK inhibitor Y27632 to determine which is optimal for your workflow.
Neural stem cells in adherent or neurosphere conditions can be differentiated into neurons using the appropriate protocol.