I am differentiating NPCs isolated from DG & SVZ (Separately) of adult mouse brain of five weeks into neuron. However, I wish to know normally what subtype the NPCS/NSCs acquires once differentiated into neuron (for e.g. GABAergic neuron or glutmatergic neuron etc.). And also what if I would have ESCs derived NPCs? Do I have the same subtype of neuron as I will get from adult NPCs? Please kindly advice regarding the same. Many thanks.
The subtype probably depends on your specific differentiation protocol and culture conditions. In general, I would expect a mixed culture. You can get an idea of what types of neurons you have by doing some immunostainings, e.g. for VGlut 1/2 (glutamate transporter of synaptic vesicles) or VGAT (GABA transporter of synaptic vesicles).
Unfortunately, I'm not aware of any protocol that gives a pure culture of the one or the other (which does, however, not mean that there aren't any). It is possible, as far as I recall, to silence release by GABAergic neurons in a mixed culture by chronic kainate treatment, but that might also influence the physiology of the glutamatergic neurons. There was a paper suggesting that Akt signalling can shift the balance towards GABAergic neurons (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2722283/pdf/zpq13064.pdf), but I don't believe that they attained pure cultures either. Sorry that I cannot be of further help.
Hi Syed,
from mouse this protocol usually works pretty well.
Supplement NPC medium (DMEM/F12+ the factor you normally use) with 0.1% FBS, retinoic acid (1 μM, Sigma-Aldrich), and forskolin (5 μM, Sigma-Aldrich). In 7-10 days you should have a pretty high percentage of GABAergic neurons.
Hope this helps.
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