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Transfer of (live) tissue


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#1 SusieQ

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Posted 04 January 2016 - 02:21 AM

Hi all,

 

My team and I are working to set up assays in which we would like to use oligodendrocytes isolated from brain material. We're collaborating with a lab in London (we're in the Netherlands) and the plan is to have the lab in London perform the in vivo experiment, which is their expertise, and then have them send us whole brains obtained from the experimental rats. However, as we would like to isolate the oligodendrocytes from these brains in our lab in the Netherlands, a method for proper transport is required so that the tissue remains vital during the transport. In vitro experimentation is our expertise, so splitting the work in this way would be most efficient.

 

I was wondering whether anyone has any experience with this? Do you suspend the brain in its entirety in a certain buffer, do you need to perfuse them with a specific buffer prior to isolation from the animals, or is it perhaps better to isolate the oligodendrocytes in London and transport only these cells?

 

I would love to hear your input on this!

 

Kind regards,

SusieQ



#2 bob1

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Posted 04 January 2016 - 03:18 PM

Hmmm, you will probably have to do a bit of experimentation with this - I would go for the extraction of the oligodendrocytes first, as cells are a bit easier to transport (and probably easier to get through biosecurity controls etc), but the cells might be fragile and start to die during transport - so you would need to work on media/buffers for this.

 

If you go for whole brain, I would perfuse the mouse and then dissect the brain out. You would need to have a system for getting oxygen into the brain during transport, which you might be able to do with a peristaltic pump and oxygenated buffer (e.g. ringer's lactate).



#3 SusieQ

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Posted 05 January 2016 - 12:52 AM

Thanks for your reply!

 

We try to keep the duration of the transport as short as possible; probably not longer than a day. I have a hunch that isolating the cells for transport instead of the whole brain will be less of an effort, as I expect the cells can be isolated and then frozen in medium + a cryoprotective agent, because ODC precursor cells can be commercially bought as well (so that would mean there is a way of cryopreservation for these cells).

 

Perhaps, if the yield will remain high enough, we can split the hemispheres of the brains and isolate the cells from one half, while keeping the other half in one piece and put both up for transport. We could use a single brain for this, if available. Although I can imagine there to be a problem with "we'll just kill an animal and ship it's brain to see what happens".






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