C2C12: Growing cells on 3D printed wood
The Pelling Lab is one of creative exploration. My current project is to try and grow my C2C12 cells on 3D printed wood. Today, my cells reached their 19th passage (averaging at one or two a week since they came into my care on July 9th). It is in the basement lab that I am getting familiar with maker and hacker logics.
With an initial proposed patent in 1980 and the first patent handed out to Charles Hull in 1986, 3D printing has been around for a while (3D Printing Industry). Initially used for prototyping purposes in the industries, 3D printers are becoming more and more of a consumer tool. In 2004, the first open source self-replicating 3D printer had been designed. By 2007, a 3D printer under 10 000$ appeared on the market. Now-a-days, 3D printers are available for consumers under 1000$ (3D Printing Industry).
The machine we have in the lab is the MakerBot Replicator 2. A desktop 3D printer, it has 100-micron layer resolution for smooth surfaces, 410 cubic inches of printing space for whole projects, an easy-to-use menu and is built for speed (MakerBot). It works by extrusion of the thermoplastic. The filament (usually PLA or ABS plastics) is wrapped on its spool, inserted into the heated extrusion head and the melted plastic lands on the platform, layer upon layer, until the object is fully built.
I began by downloading SketchUp Make, a free software for designing 3D objects. The tools are pretty straightforward and explained in the software. Given the simplicity of my first designs, it did not take too much time to complete. The next step was exporting the objects into .stl files recognizable by the printer. This file conversion actually slices the 3D object into layers the printer can reproduce (3D Printing Industry). I carried out the design of two square: one without features and one with a ridge. It is on these I am trying to grow cells.
Once the file is exported to .stl, the next step is to open MakerWare on the lab computer. It is a software used to set up the printing task: it looks just like the 3D printer. You insert your .stl 3D objects and place them where you would like them to land on the bed of the printer. This is also the interface used to specify the printer settings such as material, melting temperature, resolution, density, and if there is need for rafts or supports. This new file is then exported to the SD card which finds its place in the MakerBot. With the flip switched on, I begin by replacing the PLA filament with the wood filament. This requires unloading the PLA by heating the head and pulling the filament and simply reloading the printer with the wood filament. Choosing the right file from the “Print from SD Card” menu is a breeze and the 3D printer can finally get to work!
Costing around 40$ per kilogram (SainSmart), the wood filament is 40% recycled wood and 60% binding polymer. It is printed just like thermoplastics but gives a wooden-like appearance and smell to your 3D printed objects. It can be printed between 175°C and 250°C, with the shade of brown becoming darker as the melting temperature rises (3Ders, 2013; Francois, 2012). This property can be taken advantage of to give the effect of real wood with a tree’s growth ring! This filament was created by Kai Parthy, an avid inventor who has mixed and tested filament mixtures right out of his garage (Molitch-Hou, 2014)! In true maker fashion, Parthy reports: “I made my first blending with a hot oven plate and a cordless electric screwdriver. I pressed a hot melt into a waffle iron that happened to have 3 mm long grooves And I would feed these sticks into my Rapman printer”. While enthusiasm towards 3D printing technology is usually grand, Kai Parthy offers a moderate view on the question:
“When 3D printing first took off, one claim was very popular: ‘This is a self-replicating machine.” You know, this machine can only produce some plastic parts for itself, no screws, no electronics, only plastic. But the (wrong) claim is living on in the common mind that 3D printing may produce everything in the future. Sorry, it may not!”
Regardless, 3D printing may have a global impact. Manufacturing methods may change as this additive technology reduces the need for tools, labour and is more efficient in terms of wasted materials. Local manufacturing for specialized parts could replace large-scale manufacturing and distribution. 3D printing also offers customisation and complexity at no extra cost. 3D printing is also sustainable, using up to 90% of standard materials, is an energy-efficient technology, produces less waste being an additive technology and could reduce pollution associatied with large-scale manufacturing (3D Printing Industry). The fields of medicine, aerospace, automotive, jewellery, art & design, architecture and food all benefit greatly from the use of 3D printers. Researchers have even been able to 3D print neural cells and form cellular networks with bioink (Butler Millsaps, 2015). Most of all, 3D printers will allow consumers to take control with easily accessible designing and printing (3D Printing Industry).
So what will I be doing with my 3D printed squares? Try to grow cells on them, of course! The hope is the C2C12 cells I’ve been taking care of in the last few weeks will be able to grow on these wooden squares. The protocol is quite simple. I proceeded to split my cells as usual, but kept the remainder cells instead of washing them away. This action alone meant a great deal: these living organisms are serving a purpose.
This interaction between us feels strange at times. They’re the ones doing most of the work here, not me. Throwing them away at the end of the standard splitting protocol feels like I’m just wasting some lives. Today, I got to gather up these living organisms in a tiny drop that landed on my squares.
After an hour, I added more media and they are now sitting comfortably in the moist humid box that is their home, the CO2 incubator. They will, or will not, grow on this new surface which hasn’t been tested in the lab before. Results are unpredictable. It will be a few days for the cells to grow, then we’ll be able to track their progress through imaging. A petri dish sits in my SketchUp work board, a wooden 3D printed version of It will likely be subject to receiving cells shortly.
And now, I wait, to see if this life I’ve maintained will resist this new milieu.
3Ders (2013). Wood filament LAYWOO-D3 suppliers and price compare. Retrieved from http://www.3ders.org/articles/20130204-wood-filament-laywoo-d3-suppliers-and-price-compare.html.
3D Printing Industry. The Free Beginner’s Guide to 3D Printing. Retrieved from http://3dprintingindustry.com/3d-printing-basics-free-beginners-guide.
Butler Millsaps, B. (2015). Researchers Are 3D Printing Neural Cells Of the Brain, Unlocking New Secrets of the Complex Organ. 3DPrint.com. Retrieved from http://3dprint.com/86557/3d-printed-brain.
Francois, J. (2012). Review: printing with wood filament. Retrieved from http://www.tridimake.com/2012/10/review-wood-filament.html.
MakerBot. Brooklyn, NY. MakerBot Replicator 2 Brochure. Retrieved from http://store.makerbot.com/replicator2.
Molitch-Hou, M. (2014). The Journey of Laywoo-D3: An Interview with Wood Filament Inventor Kai Parthy. 3D Printing Industry. Retrieved from http://3dprintingindustry.com/2014/02/10/journey-laywoo-d3-interview-wood-filament-inventor-kai-parthy.
SainSmart. SainSmart 3D Printer Wood Filament. Retrieved from http://www.sainsmart.com/sainsmart-3d-printer-wood-filament-1-75mm-1kg-dark-brown.html.