Decellularizing tissues and organs

[virginiajim]

Has anyone investigated a simple DIY method of decellularizing, say, a
chicken heart or liver? This looks like it would be a great high
school biology project to introduce students to the extracellular
matrix and on into regenerative medicine or tissue engineering. I see
that PEG (polyethylene glycol) is the solution that removes cells, but
that's all I've got so far.

[Ethan]

That sounds like an interesting project. Doing a quick search, I came
across this article: http://www.sciencedirect.com/science/article/pii/S0142961206001682
It seems to cover a lot of options. To me, it looks like the most
readily available DIY methods would be osmotic shock, chelating agents
(such as EDTA or even citric acid), and maybe ionic detergents or
proteases (though SDS seems to be a bad option and they don't report
on some of the more commonly available proteases)

[Marc Dusseiller]

hoied mitenand,

in the pelling lab they are showing some DIY biohacking methods to do
decellularization:
http://plasticbiology.net/tagged/Decellularization/chrono

seems kinda easy, 6 hours in a solution of SDS (Sodium dodecyl
sulfate).

why you think its a bad option?

best,
m

[Ethan]

Based on the article that I linked, SDS has a tendency to disrupt
collagen (though not remove it from the matrix) and remove GAGs. I
would imagine that this would significantly weaken the structure of
the ECM and hinder the repopulation by cells. Then again, if this is
just for demonstration purposes, that is likely not a terribly big
issue.

[virginiajim]

Thanks. Good feedback and leads. I did a little more checking, also,
prompted by Ethan's link. Much available material requires a
subscription to access, but one link to a proposed patent was
bothersome by the scope, so I searched for "decelluarized patents" and
got over 70K hits, like this 2009 example: http://www.faqs.org/patents/app/20090202977
Anyway, I have enough to go on now, but any student wishing to pursue
the science probably needs to study patent law if they hope to apply
their results.

[Nathan McCorkle]

I would think proteases would mess up the ECM, since its protein but
not really touch the cells themselves... whereas SDS would disrupt the
cell membranes and some protein structures, but I think the protein
structures would be covalently bonded and if so wouldn't be dissolved.
Low concentration SDS with rinse cycles between sounds like a good
middle route, low enough SDS conc that the proteins aren't affected,
many rinses with fresh SDS solution to ensure all cell membranes are
dissolved.

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Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

[CodonAUG]

If memory serves the lab that recellularized a mouse heart and made it
beat used SDS.

[Ethan]

I would imagine that proteases are used in lower concentrations to
help solubilize various cellular components. It is likely a balancing
act between achieving that goal and just chewing up the ECM, though I
imagine some proteases have a higher affinity to do that than others.
Doing a bit more searching, it does look like SDS is pretty widely
used.

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