Anyone thinking about bioprinting with reprap yet?

The funny thing is that custom DNA strands are currently easily automatically synthesized. The fun bit is putting a cell around it and kicking it into action.

If you can get that bit right the next bit is bio devices that will self fabricate. (Bacteria and Yeasts are good models to start with)

DNA is routinely sequenced up as primers etc in bio labs and the technology is about ripe for a low cost DIY machine. (no cnc required just squirty things and controlled heating.)

The biggest obstacle to kitchen biotech is ignorance.

Knowledge transfer is needed to change this.

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Necessity hopefully becomes the absentee parent of successfully invented children.

Has anybody considered using a bloodlike fluid as scaffolding and nutrient delivery syst
em?

... yes, this is a regularly approach when tinkering with living cells ...

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Viktor
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Aufruf zum Projekt "Müll-freie Meere" - [reprap.org] -- Deutsche Facebook-Gruppe - [www.facebook.com]

Call for the project "garbage-free seas" - [reprap.org]

Apparently lab based cell/tissue culture routinely uses something called Foetal Calf Serum as the bloodlike fluid containing nutrients.

This is kept in the temperature and gas controlled atmosphere of an incubator oven thing I think.

I guess the name may give away what it is ..... Nice.

the problem appears to be two fold.

1. Gas and nutrient transport provision.
2. Waste Gas and effluent removal.

What I would like to know is what the organism is that produces quorn. I think it is a yeast like organism but have no idea what. Home Mycoprotein production may be doable and a good first step.

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Necessity hopefully becomes the absentee parent of successfully invented children.

Quorn is made from a fungus, Fusarium something or another, that is in the mushroom family.
I think it only occurs naturally in Britain or maybe it was Ireland.
Of course the marketing people say that Quorn actually contains the EU patented mycoprotein
and I think the patent expired in 2010 so there may be more information about it online.

I will have a rummage and see what it turns up.

I wasn't aware that the quorn organism was engineered. Therefore patentable. But is would be good news that the patent has expired. It's open season now then .....

The production process etc maybe patentable though.

I think patents etc are overdue a shakeup. There is far too much use of patents as an exploitative tool. Not really what they were intended to do.

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Necessity hopefully becomes the absentee parent of successfully invented children.

I don't think Quorm is really engineered. I think the patent covers the process to mass produce the fungus.

This would be absolutely PERFECT! You see, I have this awesome metallic humanoid robot. The problem is that it looks like a robot and I would need a living tissue skin to help disguise it. Oh, and if I ever do any time traveling, everyone knows only living things can go through time...

Cells have the ability to build themselves.. Why assemble an organ with external means when you can just.. induce the organ formation by feeding stem cells the right signals. All we need then is to know what those signals are and replicate them. No need for 'reprap' in that case. Sure it might take longer but im not convinced that printed organs will work at all. Organ structures are a consequence of the way they naturally develop.

Thus, what would seem like the most natural way to form an organ is to make one develop.

... this is one of the main goals, bio- and gen-engineering try to achieve since the discovery of DNA/RNA and the first glimpses on the applied 'functions'.

With the capability to 'program' a cell to change the internal behaviour to produce a new organ you'll be able to do much more!

With the correct 'magic words' you can heal any sort of cancer ... repair the telomeres at the ends of the DNA-strands, so the cells (and the complete body) will stay 'young' for ever ... a lost limb will grow anew ... and some more miracles of this sort ...

So if you get a safe methode to grow a complete organ, you'll have solved the puzzle all the genetics worldwide are working on

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Viktor
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Aufruf zum Projekt "Müll-freie Meere" - [reprap.org] -- Deutsche Facebook-Gruppe - [www.facebook.com]

Call for the project "garbage-free seas" - [reprap.org]

VDX Wrote:
-------------------------------------------------------
> ... this is one of the main goals, bio- and
> gen-engineering try to achieve since the discovery
> of DNA/RNA and the first glimpses on the applied
> 'functions'.

SO my point is: Why waste time trying to print an organ when you can instead spend time working on the lofty goal of growing one? The challenges in printing a successful organ and growing one from scratch are very similar, except one of these is profoundly more useful. I have this feeling that the concept of organ printing is just a result of bioengineers going amok. Feel free to prove me wrong but just from hearing a bunch of these people talk it is clear to me that they aren't really approaching the problem holistically; they use cells as a building material instead of treating them as individual organisms, to the detriment of the resulting 'organ'.

THe fact is you can't just plop a cell down in most cases and expect it to work. YOu still need transcription factors and other morphogenic signals, which is exactly what you would need if you were growing the organ from scratch. Cells can move themselves to the right places given the right signals. It just seems way too complicated to me to have a massive printer to position the cells if they are already capable of it themselves. We just need more biologists on this problem and less engineers.

As an engineer who is working on the cell printer issues, I can say we can print some viable organs today. I don't think anyone has grown a viable one yet. At our current level of knowledge both attempted printing and attempted growing are still adding more and more knowledge to what we (as humans) know about cell and organ dynamics. Have us "organ printing" people gone amok? Maybe, but printing is currently cheaper than growing and unfortunately it is money that is driving the science.

In the long run growing organs may be a better solution than printing them, but we are still taking baby steps in this field so as long as attempted organ printing keeps giving us new information, then it will continue.

Do we need more biologist? Yes.
Do we need more engineers? Yes.
What do we really need? An educated population that realizes you don't have to kill babies to do cell or stem cell research.

I suppose some growing methods could need a printing first step too.

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Most of my technical comments should be correct, but is THIS one ?
Anyway, as a rule of thumb, always double check what people write.

... for printing you need only a methode to place particles/material/cells on a surface or in XYZ ... for growing organs you need the capability to 'talk' to cells and/or changing the genetic algorhythms inside the cell core and apply some methods for quality control, so you'll skip all errors or incorrect samples.

This capability to 'speak' with living cells is some big steps away from our actual knowledge base.

Imagine what you need to change the genetics of a walnut, so it won't grow up to a tree, but to a complete house with wooden interieur

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Viktor
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Aufruf zum Projekt "Müll-freie Meere" - [reprap.org] -- Deutsche Facebook-Gruppe - [www.facebook.com]

Call for the project "garbage-free seas" - [reprap.org]

criswilson10 Wrote:
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> What do we really need? An educated population
> that realizes you don't have to kill babies to do
> cell or stem cell research.

I agree with everything you said. I suppose I was going a bit overboard with my comments about engineers taking the science the wrong way. I think what I want to say is, building or printing organs is the more obvious way to think about the process of producing new organs. However, growing organs is the more natural and thus (in my opinion) the better way. Now here is a thing: [www.bbc.co.uk] Instead of printing with cells, you print a scaffold that cells grow on. Sort of a compromise between the two ideas, but probably better right now than either one.


VDX Wrote:
------------------------------------------------------
>... for printing you need only a methode to place
>particles/material/cells on a surface or in XYZ ...
>for growing organs you need the capability to
>'talk' to cells and/or changing the genetic algorhythms
>inside the cell core and apply some methods
>for quality control, so you'll skip all errors or
>incorrect samples.

Disagree, you definately need to "talk" to the cells when you print them. And in any case you need to change their (epi) genetic makeup so the cells are stem-like in order to be able to be part of the organ.

... there were some experiments with cells floating in solvent/nutrient fluid and printed with a sort of soft-bubble-jet.

They managed to bring most of the cells unharmed to the surface and let them settle there, but more statistic displaced than in rows or grids.

Another approach is to print some structures on a thin sheet of biodegradable material and put them in the incubator; floating cells starts to populate specified regions; then you can stack this sheets to a structured 3D-object.

But this all is not 'talking' to cells ... its more hoarding and/or forcing them to specific positions on a surface ... and hope, that they will stay there ...

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Viktor
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Aufruf zum Projekt "Müll-freie Meere" - [reprap.org] -- Deutsche Facebook-Gruppe - [www.facebook.com]

Call for the project "garbage-free seas" - [reprap.org]

Re the nature v nurture debate (or growing it v engineering it).

There are issues both ways as I understand it.

Nature ie growing it. Given the time it takes to grow a person to full size, if it takes a similar timescale to grow an organ the needy recipient is likely to be too dead to receive it. Growing requires more intensive support and nurturing than engineering. Particularly as it is likely to be a lengthy process and the needs of the organs development must be catered for as must protecting it from the pathogens and forces of entropy that a fully developed body is able to fight of on a routine basis.

Nurture ie engineering it. Organs have some big obstacles to be overcome before it is possible to do this (it affects the nature argument above significantly too) to sustain a major organ you need a major delivery and waste disposal network, this is usualy know as blood vessels, blood and the ability to provide nutrients to this system and exrtact waste.

Given the above two oposing methods and constraints. Engineering arguably has the potential (maybe using scafolding) to allow the construction of organs (including blood vessels or pasageways that become blood vessels) quickly enough to benefit the recipient (always a key factor) and for the organ to survive its instantiation.

I am not sure I have anything useful to contribute to the ongoing research in these areas, I guess principaly as the folk doing it are well out of my biology-league, but can apreciate what it entails.

If growing an organ outside of a supporting, protecting and equaly developing body were as easy as sugested, I think it would have been done by now.

Thoughts for what they are worth.

aka47

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Necessity hopefully becomes the absentee parent of successfully invented children.

>"Disagree, you definately need to "talk" to the cells when you print them. And in any case you need to change their (epi) genetic makeup so >the cells are stem-like in order to be able to be part of the organ. "

Nope.

With cell jet printing we harvest cells (coring needle) from the organ that needs to be printed. We separate the harvest into the different
types of cells within the organ, throw out the bad cells, and then incubate the good cells and force them to multiply. Once we have enough cells we "print" them out along with nutrients and cell binder solution onto a scaffold in a calculated pattern. Add a little bit of electricity to the scaffold and the cells will join together and form a layer of tissue. We then repeat as many layers as are needed.

I guess the cell binder and electricity could be considered to be "talking" with the cells, but most researchers consider it to be an environmental change that makes the cells link together.

Cris

On the notion of talking to cells, (Sorry if my understanding is a bit weak here) I am given to understand that cells actualy automaticaly talk to each other and their relative positions to other cells that they "hear" stears their type and function.

Stem cells for example are initialy neutral but change to type based on what they hear from their neighbouring cells.

The talking is of course chemical signaling of some sort. And the phrase talking presuposes a lot more structure than I am sure actualy goes on. Probably "influenced by the presence of molecule X is a bit more accurate".

Following the above the implication is that by taking cells which are already diferentiated into certain tissue types the talk that they talk will influence developing cells and stem cells placed next to them.

I think Viktor is thinking about specificaly directing a cell by direct chemical communication rather than placing cells that already talk the talk next to each other for crowd control.

Although having said the above, it is perhaps as valid a way of stearing the cells communication as any other. So perhaps is valid as a method to direct the cells development and qualifies as talking ??

Cell level immortality (ie will reproduce for ever rather than dying after x many generations) is already doable and I am given to understand used regularly to keep useful cell lines for study. I guess by definition this could be considered talking to the cell's ??

I guess there are going to be other examples that I am completely unaware of too.

For me the bunch of cells that stand out as being realy odd (as in savagly different fromt the rest) from my understanding are neurone's in that they don't reproduce as such, grow very little, grow to ridiculous proportions (compared to other cells) etc.

Perhaps then having thought a little about this a combination of both Nature & Nurture ie a hybrid approach would (Maybe actualy already is) yeild best results.

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Necessity hopefully becomes the absentee parent of successfully invented children.

nice one.

Here are some companies that are doing just this sort of stuff:

Organovo(http://www.organovo.com/)

We could try to model the extruder after them. I will look through some of their papers to see if I can glean any useful information.(http://www.organovo.com/science/publications)

Matmo Wrote:
-------------------------------------------------------

> Organovo(http://www.organovo.com/)
>
> We could try to model the extruder after them. I
> will look through some of their papers to see if I
> can glean any useful
> information.(http://www.organovo.com/science/publi
> cations)
Matmo, are there requirement specifications for extruders and tolerances for the x/y/z movement? I supose those 4 geometrical parameters should define a kind of "3d printing resolution" probably significant for tissue printing.
Sorry for a stupid question, I am new in tissue engineering.

Thanks!
George

Edited 1 time(s). Last edit at 01/20/2013 02:38PM by georgep.

Dear Matmo

From my experience in the field of tissue engineering (bloodvessel like tissues as well as some knowledge of bones and cartilage) I can tell you that printing tissues (maybe later organs) will not be possible in near future. There are a lot of considerations before you can try to start to build them.
Form the technical point regarding the 3d printer, it should not be the problem, but the main problem is the biological part, which cannot be solved so simple and will be the challenge of the next decade. To cultivate tissue-like structure, cells have to be cultivated in specialized bioreactors, which supports the cells. People try to develop the bioreactors specific to the tissue, because different tissues need different conditions. Depending of the tissue, different cells are needed (in the human being you have approximately 200 different celltypes). For example if you want to make a blood vessel some cell, which would be necessary are fibroblasts, smoothmuscle cells, pericytes and endothelcells...and some others. a very simple approach would be for example using fibroblasts and endothelcells. The next problem is from where do you get them from and can you cultivate them in such quantities (access to a cellculture lab) and by the way the regulatory stuff

The biggest problem in tissue engineering prob. is another one. For most tissues you need a functional vascular system (supply system for nutrients and oxygen inside the tissue). Currently it is not possible to mimic this system adäquatly. By diffusion only tissue with a size of 50-100µm can be supported with nutrients, above this limit you need a capillary system otherwise your cell in the tissue will become necrotic....that probably is the reason why tissue engineering has great perspectives (since the beginning 10-15years ago) but only a handful products could reach the market level...(ones with quite simple fabrication)

Next...would be chemical considerations (material [collagen, hyualorn, fibrin....] biocompability, degradation, fabrication)...for using a printer you would need a material which is biocompatibly(toxicity), which could be fabricated in this temp. frame 4-37°C, which gets from a liquid state very fast in a solid one...physical considerations (temperatur consideration for cells short term 4°C-37°C longterm~37°C)...sterility of used components...and prob. one of the biggest the money...

As you see in this short statement it is a really complex issue...

if you have question feel free to ask...

volker

... I think, first practical use of 'printing tissues' would be 'artificial meat': [www.3ders.org]

Here you can undergo much complexities by growing/printing thin slices and only shortly before dispatching them as 'steak' (or in other forms) they will be cutted into the needed 2D-form and stacked/laminated to thicker and/or 3D shapes

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Viktor
--------
Aufruf zum Projekt "Müll-freie Meere" - [reprap.org] -- Deutsche Facebook-Gruppe - [www.facebook.com]

Call for the project "garbage-free seas" - [reprap.org]