Re: [DIYbio] Re: Animal slaughter -- keeping it alive in a petri

I found this (1) paper on measuring action potentials of a pig heart.

They basically just mixed a 50:50 blood and a salt sultion called
Tyrode's Solution.
en.wikipedia.org/wiki/Tyrode's_solution

A few other papers also used similar balanced salt solutions, along
with 10% serum and maybe some antibiotics/antimycotics.

I got some MgCl2 _CaCO3 tablets and monobasic sodium phosphate saline
solution (has about 1/4 as much dibasic in it too) from the pharmacy.
I hope it'll approximate the Tyrode's solution well enough, I'll just
use food store NaCl and KCl and some dextrose I have too.

Then I plan on spinning down some blood at high-speed and mixing it
50:50 with the salt solution, then filtering through a 0.4 or 0.2
micron filter. Rinse the heart chunk a few times with sterile saline,
add to the serum+saline, and hope for the best.


1
http://xa.yimg.com/kq/groups/10355996/932940578/name/ST_TQ_during_ischemia.pdf

2
The Early Detection Research Network (EDRN)
Standard Operating Procedure (SOP) For Collection of Serum
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2655764/bin/NIHMS94453-supplement-1.pdf

On Sun, Oct 13, 2013 at 9:24 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
> Patrik, there are some great pics in there!
>
> Do you happen to know if there's a 'before' pic in this arrangement?
> http://www.flickr.com/photos/yeahsnos/10006833654/sizes/o/in/pool-2303889@N24/
>
> What were you going for with the proteases? Wouldn't that destroy the
> ECM, the scaffold?
>
> I found some PBS (phosphate buffered saline) on amazon.com for
> ~$5/liter (when bought in 10L quantities)... but I think I'll just
> make up some sterile isotonic salt solution with NaCl and KCl from the
> food store (for rinsing the heart chunk of blood). Not sure how long
> the cells would survive just in saline, though I haven't looked into
> easy/DIY growth media. I figure I could spin down some blood and
> collect the plasma, but I think I'd still need to filter that to get
> 'serum'.
>
> Maybe it will be enough to keep the cells happy for a day or two... I
> seem to remember my chicken explant and trypsinized culture started
> beating within a day or two.
>
> On Fri, Oct 11, 2013 at 1:18 PM, Patrik D'haeseleer <patrikd@gmail.com> wrote:
>> Hi Nathan!
>>
>> Just did some experiments in decellularization two weeks ago. We were going
>> to do a whole pig heart, but the heart we'd gotten was missing to much of
>> the vasculature to do the perfusion.
>>
>> We did get to try some health food store digestive enzymes on heart slices -
>> both bromelain and papain worked pretty darn well. We didn't try to grow any
>> cells on them afterwards, of course...
>>
>> Here's some pics - still need to add a bunch more:
>>
>> http://www.flickr.com/groups/2303889@N24/pool
>>
>> Patrik
>>
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>
>
> --
> -Nathan



--
-Nathan

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