Where to buy nerve tissue?

[coolcash2004]

Anyone have experience working with nerve tissue?  Where can you buy it?  Any major challenges I should be aware of?  I'm trying to adapt an existing procedure for culturing bacteria to instead culture the bacteria on nerve tissue.  Thanks again,

[coolcash2004]

I did find this source though: http://www.origene.com/tissue/tissueSearch.aspx 

[Reshma Bhatnagar]

"I'm trying to adapt an existing procedure for culturing bacteria to instead culture the bacteria on nerve tissue"

Why are you trying to grow bacteria on nerve tissue? Seems wasteful and expensive. If culturing bacteria is your concern, there are several media that can be used and are widely available. At the end of the day, the nutritional requirements of the bacteria need to be met in order to culture them, and I don't see any special reason to use nerve tissue to culture any bacteria.

What is the goal for culturing your bacteria?

If you are attempting to do so to explore treatments of bacterial infections that affect the brain by culturing bacteria on nerve tissue and using drugs to kill the bacteria, the attempt would be meaningless because:

1. Both need to pass the blood brain barrier in living organisms
2. Antibiotics have been well studied anyway
3. Organisms may be resistant to the antibiotics you test

If you want to go ahead anyway, I suggest buying offal/animal brains from a butcher. I will however, reiterate that it seems pointless. Perhaps, if you could outline what you propose to do, I can come up with a more helpful reply.

[coolcash2004]

I'm trying to culture persister borrelia on peripheral nerve tissue so that I can submit the bacteria cells for proteomics analysis.

[Nathan McCorkle]

I'd look into co-culturing with something like one of these
devices(SIMpore cytoVu, produced by ibidi):
http://www.sigmaaldrich.com/catalog/product/sigma/smp0005?lang=en&region=US

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--
-Nathan

[Reshma Bhatnagar]

Here is a great review on choosing an appropriate strategy for co-culturing your cells. Nathan's suggestion of using CytoVu is excellent if you can afford it.

If you'll be co-culturing, my suggestion on animal brains is mostly out, as they will only act as a nutrient medium, and won't help much with your work if you intend to study cell-cell interaction/work on proteomics in the context of nervous tissue. The alternative is you sit with a butcher as they do their business -- not a particularly enticing prospect. Even if you do go ahead with the latter, keep in mind that culturing non-foetal neural cells are notoriously difficult, though you may have more luck with glia. Here is a protocol on culturing neural cells but I have never tested it and can't vouch for it. Here is a protocol for Schwann cells (haven't worked with them either).

Human peripheral nerve tissue will be pretty hard (if not nearly impossible) to find. You'll get cell lines easily enough (recommended). They will be expensive, though. Here are some links:

ThermoFisher
SigmaAldrich

All of these considerations will affect your model.

Do note that you'll need to maintain sterile conditions etc. Contamination will wreak havoc with your experiment.

[coolcash2004]

Thanks for the info and reply.  It is suspected that persister Borrelia burrows deeply into nerve tissue - so I'm not sure if using cell lines would reflect the environment as well.  I'm beginning to think that I might have to do this without the nerve tissue first due to cost.  Other than buying the actual tissue, which I can afford, would there be any other major costs with doing the culture on tissue vs in a medium?  I understand that contamination and failure to culture could pose the biggest risks?  

Thanks again,

[Nathan McCorkle]

It might be interesting to compare the proteome and transcriptome of
the bug grown by itself (if it can survive without nerves), and then
grown co-cultured. Then again with bugs post-infiltration of the nerve
culture.

SIMpore sends samples, if the pore size is too big (big enough for the
bugs to get through to the nerve culture side) then they can also
supply windows with smaller holes, but you'll need to setup your own
culture environment (you can get away with molding some (relatively
pure, as in PDMS, though hardware-store grade natural/clear might also
work, though I expect there might be conditioners that would leak
diffuse into the culture medium) silicone with bits of removable
plastic in a glass petri dish, probably).

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[Nathan McCorkle]

I wonder is C. elegans would work.. it has nerves... ?

--
-Nathan

[coolcash2004]

Would something like this work / be culturable?  It seems relatively affordable and its a peripheral nerve tissue.  http://www.origene.com/Tissue_Section/CS631924.aspx

Should it be significantly more expensive to culture with tissue vs without?  Other than the risk of infection / having to start over, could this be relatively straight forward?  Its easier for me to just do without tissue but I don't want to miss an opportunity that could have been relatively straightforward to do.  Thanks,

[Reshma Bhatnagar]

Would something like this work / be culturable?  It seems relatively affordable and its a peripheral nerve tissue.  http://www.origene.com/Tissue_Section/CS631924.aspx

Unlikely.

Tissue sections are mainly used for visualising pathology etc. They are dead -- most of them will be stained and preserved. Their role is more educational than anything else.

Instead, here are a couple of ways one could go about this project:

Brute force:

1. Culture the bug by itself on conventional media (as Nathan suggests): Gives you an excellent insight into how the bug works, its nutritional requirements (if you undertake optimisation studies of the media), products of metabolism etc.
2. Attempt culture on dead animal brains: Comparison with 1 to show whether the chemicals in brain matter can affect the growth of the bug.
3. Culture animal tissue and co-culture the bug: Comparison with 1 and 2 to check whether living brain matter affects the growth and metabolism of the bug.
4. Run proteomics/transcriptomics analysis for all 3 (latter will likely be cheaper, overall)

Optimsation:

1. See if the complete transcriptome of the bug is available. If so, on what medium was it cultured? That will help you even if you intend to go through the brute force route by eliminating point 1. Assuming transcriptome/proteome is available:
   
2. Identify pathways that you may be interested in terms of the mechanism of attack.
3. Culture the bacteria and isolate the protein/metabolites in question. Be thorough about the literature bit, it will save you a lot of work and money. You can run Westerns on the proteins that interest you and chuck the (expensive) proteomics/trasncriptomics.
   

I wonder is C. elegans would work.. it has nerves... ?

C. elegans would probably work -- the bug is a bacteria and they generally aren't very fussy. However, would an in vivo model of C. elegans be more insightful than culturing on dead mammalian brains? How C. elegans translates to humans for this particular problem will have to be considered.

Just looked, transcriptome of B. burgdorferi has been identified in non-human primates:
http://www.ncbi.nlm.nih.gov/pubmed/14671329

I think you can start with an informatics based approach from free resources online and then choose the path you wish to take with this. Also excel/spreadsheet is your friend.