We took our market research, and all of the work we have done previously to improve our project, working on the data we collected from the surveys, the interviews and the data extraction. Moreover we took in consideration, the professional feedback we received from the fablabs and makerspaces owners to improve our biomaterial 3D printer design every step of the way.

Aside from the market research, we seek advice from professionals linked to our project and interviewed them to discuss our project and asking them the right questions.

Finally, we produced a study about fundamental research, to understand how our work would impact various scientific field, expanding from synthetic biology.

Fablab and Makerspace community

We decided to enter and discover the makerspace and fablab community by investigating first-hand as we went to more than 10 fablabs and interviewed the owners as well as the regular customers. We conducted the interviews with the same standard questionnaire in order to have valid data we can work on to make our market research.

We shot over 4 hours of footage, and reduced it to a video of 18 minutes, that summarize well the challenges facing the Fablab and Makerspace community as well as the use of a 3D printer.

The fablabs and makerspaces we visited are all spaces providing shared machines for a community of users that can use them for their personal projects. But beyond this similarity, we found out that those different places were actually extremely diverse, both in terms of their global vision and their type of users. Some aim to help people manufacture objects they need, or objects they want to make for fun; others are focused on common projects and education of the general public; some are for anyone in the neighborhood, while other are just for students; some spaces are for researchers to help them with their research and experiments, and others aim to help in prototyping projects that will eventually be commercialized. This large variety in our interviews was a great value as we got very diverse and complete answers to our questions. Here are the main points that we extracted from those interviews:

FREQUENTATION
The frequentation of fablabs is very variable depending on the day or the time of the year. Some people come once for a short amount of time just to print something, while others are regular members and part of the community. Depending on the fablab, the number of regular members ranged from 10 to 100 per month.
The time actually spent on the machines is very short though, about only 10% of the weekly time — while the rest is spent on the conception, etc.

BIOLOGICAL PRINTERS VS TRADITIONAL ONES?
When asked whether biological printers would ever replace traditional ones, most agreed that they won’t. While the idea of using only biomaterials is attractive environmentally speaking, and the use of biomaterials has been rising in the past years, it is not realistic that they would completely replace plastic and other materials in the near future. Also the cost of biological printers is very prohibitive, and when the machine is used as an educational tool for modeling and manufacturing, this extra cost is not justified since there are much cheaper 3D printers. So the general idea was rather that the two could coexist.

GMOs AND PUBLIC ACCEPTANCE
As our biological 3D printer contains genetically modified bacteria and GMOs don’t have very good publicity in France in general, we wanted to assess the interest and acceptance of the different fablab communities regarding GMOs and biotech in general.
The answers depended a lot on where we asked. In fablabs open to the general public, there was concern that something might go wrong if GMOs were used for anyone to use, without clear knowledge of the impact it could have on the ecosystem if the GMOs were to escape the machine. Some though were aware that the bad opinion of GMOs came mostly from sci-fi movies, and that GMOs are being used in research, medicine, food, etc. a lot more that most people are aware of; they thought that scientists should work more on communicating their research, its use and the associated risks to the public.
In fablabs such as La Paillasse and Pasteur where GMOs are already being used routinely, the opinion was that the public has yet to realize that solutions brought by GM organisms actually make sense, especially regarding oil, climate change, energy, toxic waste, etc.

OPINION ABOUT OUR BACTERIAL PRINTER
Of course during those interviews we explained our project and ask people for their feedback. Usually people were excited about our printer and hoped to hear more about it in the future when it would be in a more advanced stage. In the fablabs for general public rather than researchers though, they feared that past the initial excitement of having a new toy in the fablab, people might lose interest quickly if there isn’t concrete, useful applications for it.
Thomas Landrain from la Paillasse on the other hand seemed to like our project a lot because it combines AND gates and optogenetics, which are both rather robust and well characterized nowadays. He thinks such a printer, if proven reliable, could be used not only for research, but also for fun, and in workshops. Since the materials (media, bacteria) are cheap, the use of the machine could probably be DIY-ed quite easily, which would allow the community to use it and explore its possibilities.

LIMITS OF OUR BACTERIAL PRINTER
We got excellent feedback on our project, with very constructive and useful advice which we tried to take into account in the design of our printer as much as possible.

the need of an expert to operate and maintain the machine and the cell culture
the definition of the prints might be lower than regular printers, so the quality wouldn’t be as good
the ratio financial investment / actual use. It’s an expensive machine and it’s not obvious that the general public would use it a lot
the potential difficulties to extract the product from the media
the bacterial resistance to light, since they will be exposed a lot and it could be toxic to them
the ability of the bacteria to secrete in a controlled, homogeneous manner
the need to control the temperature in the machine
the applications would be few and very specific, so only a small number of people would actually want to use it.

As Gabi put it nicely, our printer is in early-stage development and still very much an object of study by itself, so it will take time and research before we can overcome the constraints above and before our printer can start being used for other people’s projects.

Safety

Thanks to our primary research, we listened to all of the concerns that was mentioned by the general public as well as professionals. Safety issue was the most important one. Therefore we integrated a cell lysis system in our design. It is easy to activate the lysis in the cell and kill it.

We wrote a safety dossier regarding our meeting with the Dr.Namorado, to insure our subject is in compliance with the EU’s Horizon 2020 ethical criteria.
Here is the link to the Safety Dossier

Interview

To improve our work, we talked to scientists and professionals that gave us a real input on our project. All of the interviews were very informative, and most importantly gave us an opportunity to explore further in detail our work, thanks to the advice of external people, giving us a new eye on the project.

Alain Le Méhauté

Alain Le Mehaute is a french Physicist, Engineer and Mathematicians who was the first person to file a patent for stereolithography, the cousin of 3D printing. Unfortunately, the company he worked for didn’t believe in the potential of the invention and withdrew the patent without his assent. Far from being resentful, Alain explained us over a Skype interview that this was just another example of the inefficacy of French hierarchy which often prioritize short-sighted and individualistic career goals over long term innovative vision. Alain also explains that, although he was conscious of the commercial potential of his invention, he wanted to develop it at the time to bring proof to his theories on fractal geometries. This interview gave us a glimpse of the potential of our control system. Given where 3D printing stands today, Medusa will probably find applications we cannot yet predict.

Andrew Pelling

Andrew Pelling is a canadian scientist from the university of Ottawa, and the director of the Pelling Lab. He is focusing on augmented biology, his dream being redefining the limits of biology. He’s currently working on making human ears using apple. We talked to him about our project, and more specifically about our biomaterials. As we are working with non polluting, biocompatible materials, the fact that we could do anything with them drew his attention. He’s really interested in DIY bio products and what we could do with them. And gave us great advice for potential applications, as well as design of the printer itself.

John Cumbers

John Cumbers is a synthetic biologist and the founder of the Brown University iGEM team. He worked for NASA Ames and now is he the founder of SynBioBeta. We talked to him about the idea of taking our biomaterial 3D printer to space, this is an exciting thought for us, as we thought that if we want to colonise Mars, let’s not start polluting right away. Although he warned us about the difficulties of growing bacteria in space. We also asked him advices about how to launch a startup because of his position as founder of SynBioBeta, and among every thing he said, one drew our attention: “Just do it”.

Dan Widmaier

Dan Widmaier is the CEO of Bolt Threads, a company working on producing silk fiber by replicating the process used by spiders. Spider silk has amazing properties such as high tensile strength, elasticity, durability and softness. We talked to him during the Hello Tomorrow summit in Paris, and discussed our biomaterials mostly, but also the cell-free system. Indeed, he was interested in our cell-free system, however he made some comments. While it is a great experimentation and prototyping technology, it is somewhat limited for mass market commercial applications.

Anne Meyer

Anne Meyer’s lab recently created an additive-layer bacterial 3D printer with a affordable modified 3D printer. Anne is interested in using her printer for both creating biomaterials with new physical properties like graphene and for research applications like studying biofilm. Our interview with Anne gave us great insight on how bioprinter will soon find meaningful real-world applications.

Fundamental research

We looked into possible applications of our 3D gene expression control in various field in biology, health, food industry and even space! We believe our tool can really bring a difference compared to existing technique, this is why we seeked the input of professionals to discuss the large number of applications.

Brain Control

Optogenetics is closely linked to neurology as activating individual neurones by shining lights allow for a fine resolution mapping of the function of the nervous system.

Developmental biology

Conditions of the microenvironment are key to determining cell fate and small changes result in large variation of the morphology of a mature organism. Although large advances have been made in the field of developmental biology thanks to higher resolution imaging, a lot remains to be understood about the underlying molecular mechanisms and signal diffusion interactions. Being able to dictate specific 3D locations of activation of genes determining developmental processes would allow a more comprehensive overview of each gene and the mechanisms of interactions between them.

Hence, we thought that by using our tools to light activate certain cells within an organism for targeted gene expression, we could observe the effect of the signal at different spatial resolutions. Furthermore, the field of developmental biology is spans all organisms and is studied in depth in a large variety of model organisms. The modularity of our tools allow for them to be implemented in many different organisms.

Going to Space

Colonising and travelling to different planets and maybe one day different solar systems is one of manhoods dream. Medusa also likes to dream big and we believe that applying 3D control in synthetic biology for space is crucial to success. To build materials in space, it would be very useful to use microbial cultures, as they could easily be transported and then grown into specific structures with lights and space resources. They would be produced in bioreactors that would shield them from hostile environments, such as the radiation and - voila! Space could be built up sustainably using Medusa and we wouldn’t have to transport large building blocks from earth. We shared our ambition with John Cumbers, who was intrigued about our project. He explained in details the different parameters that have to be taken into account if you are thinking of growing microorganism in space.

Fermentation and Bioproduction

Speeding up Microbial production in industry remains one of the main challenges in bio-technological industry. One of the problems influencing this is the lack of control of enzyme diffusion within the cell and having high enough concentration of enzymes. By applying 3D control of enzymes within a cell using Medusa’s RNA organelle, we can create local concentrations of enzymes which could vastly speed up enzymatic pathways of production leading to higher yields and faster production.

Medusa’s light control could also be applied in Industry, where different processes could be installed in a single bioreactor using modular light patterns, this would allow for more efficiency and multi-level processes to be organised into one space.

Studying Co-Cultures

Co-cultures have become one of synthetic biology’s most promising methods for reducing metabolic loads and creating large networks. To best understand interactions between two populations and spatial interactions between them, we propose using Medusa’s optogenetic control to spatially activate expression in certain areas of certain strains to best understand interactions of co-cultures. We talked to the Imperial College 2016 iGEM team about our project and our ambition to study Co-Cultures, they were very interested in our project and they shared precious information concerning their data.

Centre for Research and Interdisciplinarity (CRI)
Faculty of Medicine Cochin Port-Royal, South wing, 2nd floor
Paris Descartes University
24, rue du Faubourg Saint Jacques
75014 Paris, France

bettencourt.igem2017@gmail.com

Team:Paris Bettencourt/HP/Gold Integrated