If you are looking to differentiate the cells in a culture or slice, you can do immunohistochemistry with antibodies against NG2 (gene name CSPG4) for OPCs and myelin basic protein (gene name MBP) for myelinating oligodendrocytes. Other useful OPC and oligodendrocyte specific genes can be looked up in this transcriptome database: http://web.stanford.edu/group/barres_lab/cgi-bin/igv_cgi_2.py?lname=mbp

If you are trying to purify OPCs and oligodendrocyets, you can use immunopanning methods like those used by Ben Barres's lab. See methods in: http://www.jneurosci.org/content/34/36/11929.full

Good luck!

Anusha offers good advice.

If you mean 'identify/distinguish' (rather than separate) by microscopy within a culture, the you can use various antibodies to immunostain for different stages of oligodendroglial differentiation. You could also perform morphological analyses to assess extent of differentiation/maturity by number/complexity of processes.

Baumann, N. & Pham-Dinh, D. Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol. Rev. 81, 871–927 (2001).

Butts, B. D., Houde, C. & Mehmet, H. Maturation-dependent sensitivity of oligodendrocyte lineage cells to apoptosis: implications for normal development and disease. Cell Death Differ. 15, 1178– 1186 (2008).

Assessing functional maturity would probably require co-culture with neurons, such that myelination could be quantified.

If you need separate high purity OPC cultures and high purity oligodendrocyte cultures, then use proliferation- and differentiation-promoting media, respectively.

If, however, you literally mean 'separate', as in collect OPCs into 1 tube and oligodendrocytes into another tube. You could try to detach cells (this will damage some surface markers) and immunopan using antibodies (to separate cells), or perform flow cytometry. But oligodendrocytes will not tolerate detachment from a substrate, and you'll lose lots of cells.

Thanks. I am curious about separating mature and immature glial cell. Actually I want to use PMA according to previous literature and it will give mature cell in serum free medium. "http://journals.plos.org/plosone/articleid=10.1371/journal.pone.0045501"

My question is...is it for every glial cell? I have MO3.13 cell.  If it gives according to protocol will I need immunehistochemistry? 

Dear Monokesh, 

for mature oligos - you can use antibodies  against CNPase and MBP. For the precursor of oligodendrocytes  - Anusha is right antibodies against NG2 - is the best marker

Good luck, 

Tatyana

You could also analyze gene expression using RT-PCR for immature (NG2, Id1, Id4, Tcf4) and mature (CNP, PLP, MAG, MBP, CGT) markers. Can MO3.13 differentiate to mature stages?

Thanks everyone a lot for adding so many important advice. 

Great !!

Hi, 

I came across this article for my journal club presentation. They used MBP and O4 for matured OPCs markers.