Hi all,
I'm doing a project on drug-induced neurotoxicity but my background is in leukaemia biology, so I'm very much learning as I go along with regards to neuroscience, neural cell lines etc. The question I really want to ask is does this drug affect functional myelination of spinal cord neurons by oligodendrocytes. We hope to use iPSC to produce these two cell types eventually, but I'm wondering whether there may be scope to do co-culture with differentiated relevant cell lines. I'm culturing MO3.13 currently, which are a hybrid oligodendrocyte-like cell line, which can be differentiated to more mature oligodendrocytes, expressing a greater range of myelin proteins than undifferentiated by treating with 100nM PMA. I've also previously used the SH-SY5Y neuroblastoma line, and have read that they can be differentiated also with retinoic acid to form more mature neurons. I'm wondering whether any
Hi,
Your message was cut but I suppose you want to see myelination on axons. It depends on your question: if you just want to see whether there is any myelin just do an immuno for MAG. If you need more detailed quantitative answer (i.e.: layers of myelin etc) you have to do electron microscopy study.
Dear Victoria,
Unfortunately, as a neurobiologist, I work with primary neurons, so I can't help with the cell lines. However, I would suggest that you check the reviews in the RG link below, or further research on PubMed, to see if anyone has successfully gotten myelination of SH-SY5Y cells.
https://www.researchgate.net/post/Why_are_SH-SY5Y_cells_popular_compared_to_other_neuronal_cell_lines
I agree with Attila's suggestion above, and just start with immuno for MAG. You may spend a bit of time getting your coculture conditions set up for myelination, so you just need a quick screen to see if it works. I am sure that you are aware that there is a large body of literature in spinal cord repair and regeneration, as well as MS, that looks at the ability of different cell types (oligodendrocyte precursors, usually) to remyelinate spinal cord neurons. So it may not just be a function of the nature of the type of neuron, but also the differentiation stage of the stem cells/oligodendrocyte precursors/oligodendrocytes, that is critical for successful myelination in your cocultures.
Two recent reviews that might be helpful in thinking about how far to differentiate your oligodendrocyte cell line:
1: Plemel JR, Keough MB, Duncan GJ, Sparling JS, Yong VW, Stys PK, Tetzlaff W.
Remyelination after spinal cord injury: is it a target for repair? Prog
Neurobiol. 2014 Jun;117:54-72. doi: 10.1016/j.pneurobio.2014.02.006. Epub 2014
Feb 28. Review. PubMed PMID: 24582777.
2: Alizadeh A, Dyck SM, Karimi-Abdolrezaee S. Myelin damage and repair in
pathologic CNS: challenges and prospects. Front Mol Neurosci. 2015 Jul 27;8:35.
doi: 10.3389/fnmol.2015.00035. eCollection 2015. Review. PubMed PMID: 26283909;
PubMed Central PMCID: PMC4515562.
Good Luck!
Jill
Dear both,
Thank you very much for your replies, and sorry about the truncated message. No idea what has happened! Despite this, your suggestions/references are excellent.
Thanks once again.
Vicky
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