Nestin and Sox1/2 are not enough.

Suggested and commonly used markers for Neural Stem Cells:

1. Nestin (+)

2. Musashi-1 (+)

3. Pax6 (+)

4. Sox1/Sox2 (+)

5. Oct4 (-)

Hello Joon and Ruslan.

Thank you for your responses. I think this will help a lot!

Do you'll have specific primary antibodies (companies) for these proteins which you have tried out and have worked well? It would be great If you'll could share that.

Thank you,

Apurva.

Hi Apurva,

Are you deriving these NPCs from ESCs? It depends if these NPCs are early or late NPCs. You are unlikely to pick up Pax6 in late NPCs. I usually do Pax6/Nestin co-staining for early NPCs and Sox2/Nestin for late. If these are early derived NPCs from ESCs I would definitly do a Oct4 staining too make sure is all negative. Hope this helps.

I have added a list of antibodies I routinely use that work well.

Nestin R&D System MAB1259

Pax-6 Millipore AB2237

SOX2 Abcam ab97959

SOX1 Abcam ab87775

BW

Hi Patrick,

Thank you so much for your suggestions.

I am deriving them form the cortical tissue of E. 12/13 mouse embryo.

-Apurva.

I agree with Patrick on early vs late NPC comment. However, I think you should stain for Musashi-1 and Oct4 regardless of the differentiation stage.

Thank you all for your inputs. I will try to incorporate your suggestions. I will share what worked.

For primary cells: Nestin, RC2 (actually a Nestin variant), Pax6 without doubt if they are from the dorsal telencepgalon, Mash1 if they are from ventral telencephalon, Gfap (dorsal at least), Sox2,

Blbp, that may be enough. Neurospheres change the transcriptionsl profile so be aware.

Joon Shim:

Neuroblast: Double cortin

aNSC: Gfap, CD133, Tlx

I think that NESTIN, PAX6 and SOX2 are sufficient

Good luck

Thank you all for your responses!

Hi Apurva

A former graduate student in my lab came up with different markers that she validated through flow to identify lineage specific cells that come from NSCs. I'll attach the link to her paper. It might be helpful. She has used CD133+,LeX+, NG2-,CD140a- cells to identify NSCs.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3583360/pdf/nihms-442984.pdf

Hello, 

I use SOX2, NESTIN, OCT4 to stain IPS derived neural stem cells. 

All cells are NESTIN+ and OCT4- but not all cells are SOX2+.   

I find that cells that are "compact in colonies" are SOX2+, while the larger flatter looking cells around the colonies are SOX2-

Hi

I work on human NSPC and need to sort them in flow cytometry. There is no single marker for such selection, so I will have to use at least two markers. I did some literature search, and the combination of Nestin and Sox1 is used for its characterisation. 

Since my goal is to sort the undifferentiated cells in the culture, do you think it would be enough to use these two markers, or could one still claim that these markers are not specific enough for NSPC?

Hi Caren,

I used nestin, SOX2 and Pax 6. These have worked really well for my characterization. I think you should make these three as starting points. Although, I believe the age at which you take these NSPCs would also afftect the expression of these markers. Also, like many others have stated in this discussion, NSPCs would also be oct 4-. So I would also include that as a negative marker. 

Hello,

I am studying neural developmental stages for my new project. After reading several papers I am a bit confused about the neural progenitor cells. Is “neural progenitor cell” and “neural stem cell” identical? Or there exists some difference between them?