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:
- 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.
- Attempt culture on dead animal brains: Comparison with 1 to show whether the chemicals in brain matter can affect the growth of the bug.
- 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.
- Run proteomics/transcriptomics analysis for all 3 (latter will likely be cheaper, overall)
Optimsation:
- 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:
- Identify pathways that you may be interested in terms of the mechanism of attack.
- 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.
On Wednesday, January 27, 2016 at 8:03:41 AM UTC+5:30, coolcash2004 wrote:
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,
On Tuesday, January 26, 2016 at 9:23:12 PM UTC-5, Nathan McCorkle wrote:I wonder is C. elegans would work.. it has nerves... ?
On Tue, Jan 26, 2016 at 6:21 PM, Nathan McCorkle <nmz...@gmail.com> wrote:
> 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).
>
> On Tue, Jan 26, 2016 at 4:11 PM, coolcash2004 <davidc...@gmail.com> wrote:
>> 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,
>>
>>
>> On Tuesday, January 26, 2016 at 10:20:10 AM UTC-5, Reshma Bhatnagar wrote:
>>>
>>> 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.
>>>
>>>
>>> On Tuesday, January 26, 2016 at 5:58:32 AM UTC+5:30, coolcash2004 wrote:
>>>>
>>>> I'm trying to culture persister borrelia on peripheral nerve tissue so
>>>> that I can submit the bacteria cells for proteomics analysis.
>>>>
>>>> On Monday, January 25, 2016 at 8:48:22 AM UTC-5, Reshma Bhatnagar wrote:
>>>>>
>>>>> "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.
>>>>>
>>>>> On Monday, January 25, 2016 at 6:11:57 AM UTC+5:30, coolcash2004 wrote:
>>>>>>
>>>>>> 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,
>>
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>
>
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