Decellularizing tissue



  • Ok i think got it now haha. When it comes to biology i know almost nothing.
  • And many hours of editing later, the video documenting the whole process is finally done. LINK
  • Is there a specific brand or type of soap you would use?
  • That's damn cool. I've been reading about custom built meshes from organic materials for growing organs, but this could work out so much better. I don't know much about biology (first degree was geology) so potential stupid question: Would this process have better prospects for getting around rejection issues in organ transplants or are the things (proteins?) responsible for rejection still left over after decellularization?

    Love the bismuth geode (browsing your Instagram). I still can't get my geodes to come out how I want. I think it's an issue of them cooling too fast. My regular crystals come out nice and big because they form in a larger pan and cool more slowly, but some idiot built the AC vent to blow right on the stove, which makes controlling the temperature difficult. I need to invest in a hot plate.
  • you need pure sds or triton. The former you can usually get online very easily. 
  • @katzevonstich yup, that's why this technique is getting so much attention. You'd literally grow yourself a new organ. And it is you since you use your own cells to seed it so no risk of rejection. 

    I use a blow torch to heat my stuff up so it's pretty toasty when I pour it into the moulds. I think I'll make a video showing my process this weekend. I already have the footage anyway. 
  • So after you decellularized the organ, how do you recell it? Do you just inject it with stem cells? Or is it more complicated?
  • Here's a link to a paper going over the process LINK
  • Just finished writing an article about my time working on decellularization. 
  • This is beautiful and a great read!
  • I was actually looking this stuff up and re found the forum hah I'm going to start working on printing the structure based off of CT scans of healthy organs. And see if I can get parts printed and re cell the structure with something like you said, cow cells.
  • Yeah. Beautiful pics.
  • Can you seed it with cells from that particular organ? Or could you use your stem cells? I was thinking about the possibility of growing a new pancreas for diabetic patients. Since this isn't a "medical" study you can take many more liberties than the fda would allow.
  • judged from number of people benefiting I'd vote for liver. Alcohol and obesity damaged way too many of those all around the globe. DIY organ kits would be quite fun to see. get that second heart and other timey whimey things.
    In all seriousness I'd be very much interested in how this progresses. Are stem cells really required or would a bunch of healthy adult cells populate too?
  • Adult cells will probably work. Though I think for some cell types it would be easier to start with stem cells, seed the scaffold and then transform them into the final cell type. Like neurons for example. Would be far more mobile when they aren't all intertwined or have an axon hanging off them. I think myocytes would be the best first thing to try
  • Gotta go stem with this. If you just try normal human adult cells, they aren't going to grow the way you want. And if you use immortalized cells, then they aren't yours and they're weird.
    But you can't just scrape out some cells and keep them growing on a substrate. 
  • I was wondering if you could make a coating out of decellularized tissue then seed it with your cells for trans dermal implants that would be without the microscopic gap due to your skin bonding directly with coating.
  • I mean that's basically what we're hoping to do, but rather than worry about seeding it with cells, we're making the coating so that cells just grow into it naturally
  • @cironex But what about the tissue how do yu get it to actually coat the implant?
  • If you want to get technical, they aren't coating. They're growing into extracellular matrix. 
  • not the cells the decellularized tissue
  • The way the coating is formulated, as it dries little pockets open up that cells can crawl into and fill (basically). The "walls" of these pockets contain some decellularized tissue derived proteins and biochemical markers. When the cells get comfy in their new homes, the biochemical markers help them determine what sort of cell they're supposed to be. They adjust accordingly and eventually release their own extracellular proteins to fill in any gaps, anchor themselves and to connect to all the rest of the bodies cells. Thus implant and skin are connected, and no more gap. Least that's how it's supposed to work. The trick is in making it actually do that, hence why we keep adjusting the recipe and refining it
  • edited November 2016
    @chironex If you get the chance you should give this article a read

    They are using hydrogels as a synthetic ECM, and in some cases the native ECM actually replaces the compound as it breaks down.  They focused more on the binding of cell adhesion molecules.  I am using functionalized PEG to develop a coating and it looks like this may be able to serve a similar purpose.
  • That's a really cool paper! I think the break down and replace method would work well. Though you'd probably want to chemically anchor some modified PEG that won't break down to the thing you're coating. Then as the stuff that can break down is replaced, some of the scaffold is still anchored to the thing you're coating, while simultaneous getting mixed with native ECM. That way everything stays attached and you don't get separation once all the PEG is gone. Either way, I like the idea of doing this with something synthetic, certainly simplifies things. 
  • I'd like to hear more about your PEG if you don't mind and/or it's not classified. What's the coating for? something like this?
  • That's not a bad idea, having a layer that would act as a barrier and keep everything bound to the implant.  I've been playing around with different magnet coatings to increase biocompatibility.  I'm looking into functionalizing with anti-Fas antibody, which would trigger apoptosis of T cells and reduce immune response to any foreign object.
  • I've been gone for a little while, finishing undergrad and a year of postgrad in biotech, but boy did this topic pull me straight back in.

    @misslitty I actually gave a presentations on a very similar technology this year. There's been some amazing work done towards cardiac tissue engineering, and one of the best bits is that the hydrogel is so flexible in some cases that the whole artificial tissue is injectable. If you want a (somewhat) thorough review of that field, and the standard of care for ischemic cardiac tissue damage, over the last two decades then let me know.

    But on the offchance that people don't want to spend a few days reading outdated medical practices for heart attacks, these are the most interesting papers I found about use of artificial scaffolds for growing tissues.

    In the second one, they included gold-nanorods in the scaffold to increase conductivity, and managed to show the entire structure beating spontaneously in a petri dish. Gold nanotech might be a bit beyond what we can do here, but it certainly shows that growing things this way is possible. Also, if I remember correctly, they used stem cells or myocytes harvested from rats to grow the tissues.

    But going back to the original topic of this thread, those decellularised tissues look amazing! I'm still freaking out a little bit that you made a ghost heart!

    As has been said already, that kind of technology is a huge step towards saving a lot of lives through transplants. And once you've got that scaffold, it is 'as simple' as growing the appropriate (or inappropriate, if we feel like it) tissue in there, but mammalian cell culture has a reputation for being incredibly difficult. Basically if someone looks at it funny it'll get infected with something. If you can pull it off, though, it'll be mind-blowing.

    PS sorry for the wall of text, I'm just a little excited to be back here.
  • Im glad you liked it! Honestly it was really easy to do. I think more people should be experimenting with it. And Glims has been experimenting with doing it using regular dish soap instead of SDS and has been getting great results.

    As to the gold nanorods, those are fairly straight forward to synthesize, though you could probably get the same effect by adding a small quantity of graphene which is easy to synthesize (a kitchen blender or a small electrolysis setup is all you need)

    We'll be able to do things like the meat berries or 3d bioprinting within a year I think. We're aiming to be mammalian cell ready by the summer. It's really not as difficult as you'd think, it's more that the equipment is very expensive. Typically the solution you grow the cells in is full of antibiotics and antifungals so it's less of a problem, but you do still need really good sterile technique. 
  • So, a few odd questions.

    Is this at all edible after decellularizing?
    And would this have any calories after decellularizing?

    Could be neat to decell something and then be able to eat loads of what would likely be rather tasteless material and fill up on nothing.
  • If you want to do that, I suggest Konnyaku. Bowl of noodles has 1 calorie , it's all fiber, have some with broth and veggies and feel full all day.

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