NESTIN is expressed in the pre-neural and early neural stages and downregulated (not completely) in late neural stage. Here are some examples for late neuronal markers: Beta (III)-Tubulin (also known as Tuj1, which is the name of the antibody against Tubulin, but many use this name), NeuN, NeuroD1, Ngn2, etc. Note that these markers also expressed in early neural stages but in a smaller amount and you can observe a low expression level of early markers in late neural stages. That happens because neuronal culture is very heterogenic.

(also note that I working with mouse ESC-s, so I believe there is a little difference between mouse and human neuronal culture, but these markers are general; and I do not know which differentiation method you used)

Hi Gergő, Thanks for your detailed answer.

HI Millie,

I dont know what exactly you want to know about stages.

My suggestion for hESC-stages  in neuronal differentiation would

include: hESC-EBs-Rosette-early NPCs- commited NPCs-Neurons

There are many beautiful papers are available for analyzing each stage and getting different kind of neurons.

 But I will give you a very basic paper to analyze each of the sages.

http://www-ncbi-nlm-nih-gov.ezproxyhost.library.tmc.edu/pubmed/?term=Cho+E-G+et+al.%2C+PLoS+ONE+6%288%29%2C+2011%00

Cho E-G et al., PLoS ONE 6(8), 2011

Yeah Nestin is highly expressed in Mature NPCs. But when you are specifically looking at a one kind of neurons, for e.g glutamatergic neurons or dopaminergic neurons, you could always look into early neuronal markers for the above neurons in committed NPCs. The emergence of early neuronal markers or neuronal transcriptional factors would give an indication about purity and robustness in your neuronal cultures.

let me know if you want any specific papers. I would happy to help.

S

Hi Soumya, thanks a lot for your help!