Hello M. Yousef,

To do your experiments, I guess you are looking for an enriched culture of neurons with minimal glial contamination?

For my PhD studies, I am using a new differentiation protocol for converting human iPSCs - to enriched culture of neurons (motoneurons and interneurons together) with minimal glial contamination.

During my PhD studies I have discovered that motoneurons do not functionally mature in cultures when astrocytes are missing from the same. I agree with your supervisor that people do culture neurons without astrocytes and achieve functionally active neurons and this is the case with motoneurons as well. Though, the functional output obtained from motoneurons grown without astrocytes is nowhere near to what you achieve when the same motoneurons are grown with astrocytes. Also, the synaptic activity seen in neuronal only cultures is minimal and it only becomes prominent and reliably quantifiable when the neurons are grown in presence of astrocytes.

If the effect of agent you looking to study has anything to do with motoneuronal maturity or its function, I would not recommend using cultures of motoneurons without any astrocytes. Though, I know of other ways it could be done if you interested.

Let me know if you need further explanation.

Best

Amit

I agree with Amit. Co-culturing neurons with astroctyes significantly enhances and accelerates their functional properties (ie electrophysiological). Astrocyte-conditioned media supplemented to neuronal cultures also works well if you do not want to co-culture. Hope this helps.

Thanks everybody

This helps a great deal