Jess: Cultured Pork
Vascular Tissue Engineering and Bioreactor Design Optimization for Cultured Pork
New Harvest Research Fellow: Jessica Krieger (BS in Biology, BA in Psychology; Kent State University, Northeast Ohio Medical University)
Project End Date: October 2019
Project Duration: Full time; Three years funded
Institutes: Kent State University / Northeast Ohio Medical University
Supervisors: Dr. Min-Ho Kim (Associate Professor of Biological Sciences, Kent State University) & Dr. William Chilian (Integrative Medical Sciences Department Chair; Northeast Ohio Medical University);
Tissue vascularization is the most pressing technical challenge to scaling up the size and thickness of in vitro meat. Strategies for culturing meat that lack robust vascularization must typically resort to decreasing the ratio of muscle to culture system volume, which increases resource demand and tissue dissimilarity to native muscle. In vitrovascularization techniques have been unsuccessful for decades because they lack elements of in situ tissue form or functionality. Engineered vascular systems must include essential cell types besides endothelial cells to promote the growth and stabilization of blood vessels. Vascular functionality additionally requires technical development of a culture system capable of perfusing oxygen and nutrients to deep tissues. Jess focused on the rapid generation of blood vessel networks from an alginate scaffold that yield interconnected perfusable luminal systems. This is a top-down strategy to design vasculature of variable blood vessel diameters with the capacity to grow nourishing capillary beds that are connected to larger arterial and venous vessels. The stabilization of this anastomosed network within engineered muscle was facilitated by a perfusable bioreactor system capable of organizing and maturing muscle fibers through electrical stimulation.
Jess is conducting basic cultured meat research, looking at the prohibitive costs of production and finding cost-effective solutions, as well as using 2D cell culture and 3D tissue culture to inquire into how we can grow bigger and better muscle while retaining the taste and texture of meat harvested from livestock.
In addition to this basic research, Jess is working on a bioreactor system for culturing meat! Although we believe that in the future, cultured meat will be produced in bioreactors rather than in culture flasks, there is no bioreactor currently in existence which accommodates all of the physiological needs for growing meat in.
The bioreactor system that Jess is designing will improve nutrient and oxygen delivery to the meat cells by acting as an artificial heart that pumps artificial blood into the muscle. This system will also “exercise” the muscle to improve its development, the same way that muscle is exercised inside the body to increase its strength. The hope is that the meat cells grown in Jess’ bioreactor system will produce a bigger and better output than current known methods are able to.
A microscopic photo showing some early signs of bovine (cow) endothelial tube formation, which - it is hoped - will eventually lead to blood vessel formation if the conditions are right.
Pork myotubes, isolated by Jess!
Jess is also developing methods to isolate muscle cells from pigs. Why pigs? As Jess says, “Pigs are one of the smartest animals on the planet (along with humans, elephants, and dolphins), but also one of the most delicious (I mean, who doesn't love bacon?). Therefore making an in vitro meat option for pork and bacon is imperative."
When Jess sent us this microscopic photo, we thought it had to be from a sample of store bought pork, but nope -- she cultured these herself in the lab!
Watch this space for the updates on Jess' research!
Written by Jess Krieger and Erin Kim, December 5, 2016, with updates on July 9, 2018.
Modular Bioreactor Design
Modular Bioreactor Design
New Harvest Seed Grantees: Han Zhang, Yi-Fan Chen, and Kyle Manke at the Faculty of Applied Science at the University of British Columbia, Canada
Project End Date: May 2018
Project Duration: One year
Institutes: University of British Columbia
Supervisors: New Harvest Research Fellow Jess Krieger and New Harvest Research Director Kate Krueger
This project aims to design and build a scalable, modular bioreactor prototype for cultured meat production to be used in a lab setting. This novel bioreactor will be modular in that cells can be grown on trays with self-contained fluid circuits which can easily be switched out if goals change. The bioreactor is particularly scalable because all fluid circuits are driven by a single peristaltic pump head. Thus, the number of trays can be increased simply by adding more trays, without the need for additional pumps and complex tubing circuits.
The bioreactor case, almost ready to go!
For the purposes of this project, 8 cell culture trays will be fabricated. Each tray will hold up to 4 cell tissue chips, a clean fluid reservoir, and a spent fluid reservoir. In addition, the trays must be able to survive being autoclaved at 121 degrees celsius and 1 atm gauge pressure. The bioreactor cabinet assembly will also be fabricated. The cabinet must be airtight when sealed, and house the pump, electronics for controlling internal temperature and up to 8 culture trays.
These freshly cut trays will house the cell culture chips, serum reservoirs, and drains inside the bioreactor when complete
To maximize the chances of the tissues surviving and growing, the bioreactor environment must be carefully controlled. Key bioreactor environmental parameters are temperature, CO2 levels, humidity, and fluid flow rate. For this early prototype, only flow rate and temperature control will be implemented.
When complete, the bioreactor will be shipped to New Harvest Research Fellow Jess Krieger’s lab in Kent, Ohio, for subsequent in vitro tissue experiments. Until then, the Engineering Physics Project Lab at UBC will be the primary location for project work.
Project Relevance: The goals of this project are twofold:
1). to design and build a bioreactor that will accommodate modular tissue-on-a-chip devices to be implanted, serving an immediate research interest today, while 2). demonstrating a scalable property useful to the budding cultured meat industry. The team is collaborating with New Harvest Research Fellows working on bioreactor related projects. Like all New Harvest-funded research, results will be made available to the public.
As you can see here, the bioreactor prototype is still being built from scratch, and it has yet to be tested for cell culturing capabilities. We’re looking forward to sharing the progress of this project as it develops!
The UBC modular bioreactor seed grantee team members Han, Yifan, and Kyle
Stay tuned for updates on this and other New Harvest research projects!
Marie: Avian Muscle Cells
In Vitro Avian Myoblast Suspension for Mass Scale Production and Consumption
New Harvest Research Fellow: Marie Gibbons (B.S. Zoology, Minor Psychology, North Carolina State University)
Project End Date: May 2018
Project Duration: Full time; Two years funded
Institutes: North Carolina State University, Raleigh NC; Prestage Department of Poultry Science, College of Agriculture
Supervisors: Dr. Paul Mozdziak (Graduate of Physiology Program Director, North Carolina State University)
In vitromeat culture offers a solution to unsustainable meat production while still providing animal products to our growing population. Cultured meat research has already begun but is faced with many obstacles. Common problems include the availability of cell lines, efficient production of cells at a large scale, and cell growth using plant based media and scaffolds. This research aims to address these issues by generating muscle cell lines that can be scaled up in a bioreactor and exploring plant based materials that can be used to grow these cells. White meat is an ideal material for meat culture due to its anaerobic activity, low fat content, and plasticity.1,2 Dr. Mozdziak has been cultivating turkey satellite cells and myoblasts since 1992. North Carolina State University will derive cell lines either from existing clonal or mass cultures that Dr. Mozdziak has generated, or from primary culture. Subsequently, the cell lines will be adapted to suspension culture, while retaining their ability to differentiate into myotubes on a solid, plant based substrate. A shift from animal agriculture to lab-based products can potentially reverse global warming, decrease foodborne illnesses and environmental pollution, and spare billions of animals. This study aims to establish an efficient method of culturing avian myoblast cells on a large scale, leading to further research in the field of cellular agriculture.
Dr. Paul Mozdiak and his graduate student, Marie Gibbons, who are working on establishing the first accessible avian muscle cell line for the production of cultured meat.
Dee, Dr. Paul's laboratory technician, in Dr. Paul's laboratory at North Carolina State University.
Dr. Paul in his lab, looking at avian muscle cells.
Abi: Vascularized Skeletal Muscle
Creation of In Vitro Thick 3D Vascularized Skeletal Muscle Tissue for the Production of Cultured Meat
New Harvest Research Fellow: Abigail Glencross (MEng Chemical Engineering; University of Bath)
Project End Date: May 2017
Project Duration: Full time; Three years funded; One year optional extra for completion.
Institutes: Kings College, London; Maastricht, NL
Supervisors: Dr. Mark Post (Professor of Vascular Physiology; Maastricht University); Dr. Lucy Di-Silvio (Cell Biologist; Kings College London)
The University of Maastricht has proved from adult bovine skeletal muscle stem cells it is possible to create edible cultured meat in a self-organised cell culture. This thin tissue (<0.5mm) can then be utilised to form ‘processed’ meats. These meats are fashioned from offcuts of the farming industry, however the vast environmental and ethical impacts of conventional meat are derived from whole primary animal products e.g. steak. In order to mitigate these negative effects thick 3D tissue must be produced. This requires synthetic vascularization to successfully facilitate oxygen and nutrient flow to, and metabolite removal from, the innermost cells to prevent necrosis. Currently self-organised tissues are limited by diffusional mass transport of these elements, and novel concepts of sustaining the innermost cells of >0.5mm tissues must be addressed.
On May 1, 2017 New Harvest discontinued the research conducted by Abi Glencross at King's College London.
A variety of situational factors made this project no longer an effective use of donor dollars. Some of the reasons for discontinuing this work include 1) an ambitious project that was not meeting the proposed timeline 2) challenges of the Research Fellow addressing a more unconventional approach to tissue engineering, 3) inadequate research support in this new field from the existing scientific community. New Harvest has made several improvements to the Fellowship program based on following the progress of this project. You can read our blog post about how and why we came to the decision to close Abi's research project here.
Perfect Day Foods
Perfect Day Foods is a San Francisco-based cellular agriculture company making milk from cell culture. Isha (our Executive Director) founded Muufri (as it was then known, until they rebranded as Perfect Day in August 2016) with Ryan and Perumal in April 2014. As of 2016, they've raised over $2 million, made their first prototype, and have directed considerable interest and funding to the field of cellular agriculture through their huge press exposure.
February 2017 Update
We're a little late to the party on this one. In 2015, Mark Steer of the University of West England reached out to Perfect Day to conduct a preliminary life cycle analysis and environmental impact study on animal-free brewed milk compared to milk from cows.
Here's a summary of what he found.
While this study is preliminary, it gives us great hope that Perfect Day's process will indeed be able to mitigate the impact that milk has on climate change, and land, energy, and water use.
If you're interested, the full study can be found here.
Written by Isha Datar on February 16, 2017
April 2016 Update
For the past two years, Ryan and Perumal have been hard at work perfecting their process, growing their team, and thinking about the kind of company they wanted to create.
After lots (and lots) of late nights, they decided to change their name from Muufri to something that they felt better captured their company’s values, process, and products.
They came up with hundreds of names. And then one day they stumbled upon a study from two scientists who discovered that dairy cows who listen to soothing music like the Lou Reed song ‘Perfect Day’ are calmer and happier, which makes them produce more milk.
Then it hit them. They could make their own kind of Perfect Day. One where we could enjoy the dairy foods we love, while making the world a kinder, greener place. And, the rest was history.
They’re hard at work on research and development and will be introducing our first products in 2017. Can’t wait that long? Check out their new website to sign up for their newsletter, follow Perfect Day on Facebook, Instagram, Twitter, or say hello at firstname.lastname@example.org!
Written by Erin Kim on April 30, 2016
December 2015 Update
Muufri has their latest milk prototype ready for tasting. Tasters have so far described the milk as having the same mouthfeel as milk from cows.
The latest prototype of Muufri's milk.
Muufri has also expanded their team! Below, from left to right: Balakrishnan Ramesh (Postdoc), Perumal Gandhi (Cofounder), Han Chen (Fermentation), Lou Hom (Strain Development), Ryan Pandya (Cofounder/CEO)
Written by Isha Datar on December 30, 2015
November 2015 Update
On March 19, 2015, New Harvest suggested to Ryan and Perumal that they apply for the Dutch Postcode Lottery Green Challenge, the largest annual international sustainability innovation competition that seeks to promote a low carbon economy and a greener future by investing in emerging green entrepreneurs
Six months later, in September, Ryan was on stage in the Netherlands sharing his pitch on Muufri.
With that awesome pitch, Muufri won €200,000 in non-dilutive funding to make milk without cows. Another testament to New Harvest's ability to direct funds to cellular agriculture.
Written by Isha Datar on November 22, 2015
Perumal Gandhi holds an early prototype of Muufri's cow-free milk.
April 15, 2014
I emailed two New Harvest volunteers that had never met each other before with an idea. Did they want to start a company to produce milk in cell culture?
Perumal Gandhi and Ryan Pandya of Muufri
My friend Pantea from Synbiota had told me that a new biotechnology accelerator was seeking applicants for summer 2014 and winners would have access to laboratory space, mentorship and $30,000 in initial funding. She wondered if I knew any new start-ups that could apply. I didn’t… but I started thinking. Meat – that’s kinda tough for a few months in the lab. But milk – now that was something you could get going in just one summer.
Ryan Pandya was one of the first people I had spoken with after I joined as New Harvest’s director in January 2013. He had dabbled in cultured meat research and first brought to my attention the idea of making milk in cell culture. He was a biological and chemical engineer who had just graduated from Tufts University.
Perumal Gandhi I had met for the first time less than a month before – he sent me a message on LinkedIn in March asking advice for how to tailor his education. He had a biotechnology undergrad degree and was doing a Masters in Biomedical Engineering at Stonybrook University on a student visa. They were the only two people I’ve met in New Harvest’s network that had mentioned the idea of producing milk in cell culture.
I emailed both of them, asking if they wanted to apply. Amazingly, they did. It was a crazy time crunch. The deadline was in 4 days.
April 19, 2014
We decided to apply as the “New Harvest Dairy Project,” hoping that New Harvest’s established network would help with the application.
Ryan had already done a lot of work on proposals around producing milk in cell culture. He had recently moved to an entrepreneurial co-living space in Boston called Krash, and he got in by posing the idea of producing milk in cell culture. Over the next few hours, and days, I was just shocked and impressed to see how Perumal and Ryan burst forth with all this research literally overnight. Patent searches, protein structures, pros and cons on different steps in the production process. It was inspiring, to say the absolute least.
Four days later, after several hours on Google Hangouts, Google Docs and Prezi, we had a sound presentation. One day ahead of schedule!
April 22, 2014
WE GOT IT! The New Harvest Dairy Project was going to receive $30,000 and laboratory space for the summer. We were thrown into a frenzy of incorporating, buying plane tickets, opening bank accounts, getting visas and quitting jobs. We kept stumbling on what to call this thing. The names we came up with were embarrassing, to say the least (Noccau, Herdler, Bovino are just a sample of the atrocities we came up with). Finally we got on a call and decided on Muufri. Great searchability, a free .com, and we thought it sounded awesome. Haven’t looked back since!
The name and logo we put together for the New Harvest Dairy Project: now known as Muufri!
April 28, 2014
We incorporated Muufri.
April 30, 2014
Ryan lands in Cork, Ireland. When I first saw photos of Ryan in Ireland I couldn’t help but think to myself – “What have I done?!!?!” Seemed to turn out OK though…
May 8, 2014
New Harvest’s Dairy Project goes out in the NH newsletter. Feedback from the newsletter was fantastic. The Muufri team was already receiving fan mail and media inquiries from journalists, including New Scientist – a forefront science magazine in the UK.
Perumal (in a Muufri t-shirt we designed ourselves) and Ryan just outside the lab in Ireland
May 16, 2014
Perumal lands in Cork.
May 19, 2014
Isha lands in Cork. The Muufri team meets in person for the first time ever.
Literally a couple hours after we first met.
Meeting for the first time was nuts. I mean, we were all essentially strangers. I was shocked by how well we got along online and how well we seemed to work together remotely. But what about in person?
It turned out to be a breeze. The other accelerator teams there thought we were all old friends – not that we had just met. It was unbelievable.
Muufri’s new workspace at University College Cork, in Cork, Ireland
The following weeks and months turned out to be a flurry of experiments, research, and making contacts. Without a doubt, New Harvest helped hook Muufri up with all kinds of resources and connections.
June 5, 2014
Natalie lands in Cork.
Natalie Rubio, New Harvest’s intern in 2014, in the lab where Muufri was getting started in Cork, Ireland
During the summer of 2014, Natalie Rubio was New Harvest’s star intern. Knowing she was up for travel this summer, I invited her to come to Ireland to work more closely with me (Natalie and I were also working remotely) and help out with Muufri/take advantage of the accelerator environment and learning experience. Natalie stayed for a whole month.
June 30, 2014
Muufri’s self-authored article appears in New Scientist.
The summer proceeds with a lot of hard work in the lab and online.
Ryan adds precise amounts of milk fats to a mixture of powdered casein, powdered whey, and water. This early experiment was to see if milk could be reconstituted from its component proteins, fats, and water.
July 2, 2014
Horizons Ventures contacts Muufri.
Muufri had been engaging with several investors throughout their time in Ireland, but were especially blown away when Horizons contacted us. The disruptive investment group has a killer portfolio including huge brands like Facebook, Siri and Spotify, alongside our inspiring friends Hampton Creek and Modern Meadow. They had seen our piece in New Scientist and wanted to chat.
July 31, 2014
Ryan, Perumal and Isha land in Hong Kong to meet with Horizons Ventures.
Ryan, Isha, and Perumal in Hong Kong
This was one of the most surreal things for me. I got my ticket to Hong Kong the day before flying out, and the whole 15 hour flight I was thinking to myself: this is nuts. We were getting the richest man in Asia excited about transforming the food system. Our conversation was great and we could feel the excitement. We wanted to do something big and Horizons wanted to help us do that.
As we worked through the paperwork, I decided to remove myself as a company founder on paper. It is important to me to keep the non-profit voice strong in this new, emerging industry. It will help the industry grow faster, more responsibly, and encourage cooperation and communication. I was more than happy to put the future of Muufri in Ryan and Perumal’s extremely able hands. From then on, New Harvest held a bit of equity in Muufri, and I personally have none.
August 29, 2014
Muufri meets again – in New York City. It is beginning to get ridiculous how we’ve been meeting up in places where none of us live. This trip turns out to be more of a fun visit. We check out Modern Meadow’s Brooklyn lab, which is just beautifully outfitted with the most high tech gear – then we check out the community at Genspace and meet a lot of talented folks who can relate to our modern-day, internet-relationship start-up story.
Sarah Sclarsic of Modern Meadow, Ryan Pandya and Perumal Gandhi of Muufri, Francoise Marga and Andras Forgacs of Modern Meadow, Paul Shapiro of the Humane Society of the United States, and Isha Datar of New Harvest at Modern Meadow’s office. Ryan, Perumal, Paul and I tasted cultured meat for the first time just before this photo was taken!
We also made a little jaunt out to Connecticut to meet Ryan’s parents!
Isha, Ryan, Ryan’s Dad Sameer, Perumal, and Ryan’s Mom Nandini at Ryan’s parents home in Milford, CT.
September 1, 2014
Muufri moves to San Francisco.
And really, this is where phase two of the adventure begins. The team starts securing laboratory and office space and the search for new hires begins.
September 30, 2014
$2 Million lands in Muufri’s bank account.
And this is where it all started to finally feel real. I mean, the past 5 months and 15 days had just been an absolutely whirlwind. We were an idea – a seed – blowing in the wind. We went on an international adventure, three people from three different countries – Canada, the US, and India, meeting in three other countries – Ireland, the US and China, not knowing where we were going to land. It was the kind of whirlwind which seems like it’s gonna just blow over or blow away and be gone as quickly as it came. But seeing the money in the bank was like seeing Muufri planting some roots. We’re trying to break ground on making a food future which is sustainable, healthy and humane. This is really, truly, just the beginning.
The new lab space! Newly leased and ready to be outfitted for making milk without cows.
Written by Isha Datar on October 4, 2014.
Milk is made up of a handful of proteins (casein and whey proteins) and fats. The proteins contribute to the creaminess of milk and the ability to turn milk into cheese (aka the functionality of milk) and the fats contribute to the flavour.
Today, milk is made by artificially inseminating a cow at 13 months of age, having it bear a calf 9 months later, having the calf removed (to be made into veal), and then maintaining the cow in a lactating state for about two years. By age four, the dairy cow is culled for beef.
Milk is usually made by mother cows kept in a lactating state in an industrial setting. Instead, we can make the exact same milk by brewing it, using a culture that consumes simple sugars to make milk proteins.
Muufri’s milk proteins are made in yeast, rather than in mother cows. Yeast is reprogrammed to produce milk proteins by inserting the genes for casein and whey proteins into the yeast cells.
The yeast culture can then be grown in large stainless steel tanks to brew milk. The yeast starter culture consumes simple sugars to produce the exact same milk proteins that a lactating cow would produce. The process of yeast consuming sugar to make a different product is called fermentation. After enough milk proteins have been produced, the yeast and milk protein mixture is separated so only the casein and whey proteins remain.
Ryan warms up some milk fats in a water bath before he adds them to milk proteins and water to make milk.
The fats are sourced from plants. The proteins, fats, and water are combined to make milk. Because this milk is molecularly identical to milk from cows, it can produce cheese, yogurt, kefir and cream.
Compared to producing milk from cows, producing milk proteins from yeast offers many benefits.
First, there are zero animals involved in the process, which means there is no opportunity for contamination by bacteria like Salmonella and E. coli, which live in the gut and feces of animals, nor contamination by other common milk contaminants, such as blood or pus. Because of the reduced risk of contamination, Muufri’s milk could theoretically have a longer shelf life, and require less pasteurization.
Second, it is much easier to maintain a stable supply of milk if it is made with yeast and plant-based fats. The milk supply varies depending on environmental factors and the spread of disease. Muufri’s system is much less prone to these uncontrollable forces.
Third, Muufri is using a very simple organism - a yeast, which doubles every couple of hours - rather than a cow, which produces milk after about two years. Because we’re using a much simpler and direct system for producing milk, it should require less land and water inputs to produce Muufri’s milk. It should also produce fewer waste products and a much safer working environment for milk producers.
Using a yeast based system to produce milk also allows us to do things we could never do before. For example, because we are choosing which components to produce, we can tailor the milk mixture. For example, we can exclude lactose and/or cholesterol. We could also alter the proportions of the components to make milk that is particularly protein rich, or creamy, or flavorful.
The last time we used biotechnology in dairy, we created cheese, yogurt, and other fermented milk products. Today we’re introducing fermentation to dairy in a whole new way, and it’s going to be exciting to see what new flavors and formats these new foods might have.
Written by Isha Datar November 5, 2015
Clara Foods is a San Francisco-based cellular agriculture company making egg whites from cell culture. New Harvest launched Clara Foods in November 2014 by connecting David Anchel and Arturo Elizondo, who at the time were just two members of the New Harvest Community. What started as the New Harvest Egg Project has now raised over $1.75 million!
Early prototype of Clara Foods’ hen-free egg white meringues.
October 20, 2014
I emailed David Anchel, a cell biologist that had been keen to take animals out of the food system, if he wanted fly to San Francisco with me.
David had been in touch with me over a year. We would catch up often on how his PhD was going, how New Harvest could grow, and how he could contribute to the field of cellular agriculture. David was so committed to the cause. Culturing animal products without animals is what got him into biology.
I thought it would be catalytic to introduce him to some New Harvest folks in the Bay Area, to help him figure out what his next steps might be after his PhD ended. David happily agreed.
David Anchel of Clara Foods
November 6, 2014
David and I had just gotten to San Francisco and were heading to a food technology conference that New Harvest was presenting at.
We (David, Ryan and Perumal of Muufri, and myself) were sitting at a table at a (not so exciting, to be honest) conference, introducing ourselves and our missions. Others at the table were fairly uncomfortable with New Harvest’s work, seeing it as unnatural and bizarre. After some thoughtful conversation, they did end up agreeing that they would rather eat cultured meat than meat from a factory farmed animal, so that was progress!
As we went around the table, to our surprise, Arturo introduced himself by saying he had just written a paper on food security in China, and that the conclusion was that the Chinese government should be funding research into cultured meat. Arturo said he watched my TEDx talk and that’s how the paper’s conclusion came together.
Arturo Elizondo of Clara Foods
I was pretty shocked because I thought we were the only technology-focused foodies in the room! We ended up having dinner with Arturo and finding out more about him. He had recently turned down prestigious job offers to become a pioneer in the emerging cellular agriculture space!
I asked Arturo to come by Muufri’s lab in a few days to meet more people from New Harvest.
November 9, 2014
We get together with the several New Harvest donors, scientists and supporters in the area (a few people flew in too!) at Muufri’s headquarters for a New Harvest event.
Over the course of our trip, David joins in on several meetings, inspired by what our community is doing and our approaches to advance animal products made without animals. He’s inspired, particularly after seeing Muufri’s lab and thinking through their method to use yeast to make milk proteins. As time goes on, David begins to look increasingly wilted throughout the day – we soon find out it’s because he’s been deep into the scientific literature at night, looking at how to make egg proteins in culture. The idea of creating a chicken’s oviduct in vitro came up… but didn’t make the cut. Eggs, particularly egg whites, could be made much more directly in yeast culture.
In the last couple days of our trip, David has put together a detailed proposal of how to make egg whites, including cost comparisons, and a scientific plan.
On the flight back, the night after the salon, I wonder how we can quickly turn David’s work into a reality. We decided a fast way to do that would be through the IndieBio SF accelerator, a related-but-separate accelerator from the one Muufri went through. They were accepting applications.
But David needed a collaborator – he could focus on lab work but he needed someone to focus on the business plan, doing market research, reaching investors and creating a future consumer base. I had an idea of who that could be.
November 12, 2014
Arturo, I have an idea…
Two days after getting back from San Francisco, I emailed Arturo with an idea – would he want to join David in making eggs without hens? With New Harvest’s help and guidance, I knew we could put together a good proposal in a short amount of time.
Arturo said yes the same day. By the next morning we had already gotten a Google doc started with our application materials.
November 17, 2014
The “New Harvest Egg Project” is submitted
After 5 long days of Skyping and Google Doc-ing… we submit our detailed proposal.
Isha, Arturo, and David of Clara Foods
November 20, 2014
“New Harvest Egg Project” invited for interview
We work harder on a more detailed scientific plan to discuss with Ron Shigeta, who is doing science due diligence for IndieBio. Overall the IndieBio folks – Arvind Gupta, Ryan Bethencourt, and Ron are totally excited about our project.
David and Arturo started brainstorming what the company’s name could be. “Clara” means egg white in Spanish, Arturo’s first language. It was also the name of David’s dog!!! Clara Foods was born.
December 10, 2014
We did it! “New Harvest Egg Project” gets $50K + lab space to make egg whites without hens.
We’re moving to California!! IndieBio’s program is set up to start March 1, but we make arrangements for David and Arturo to get there ahead of time. David moves to San Fran in late January, and gets right to work on the science at Berkeley Biolabs, so we can really hit the ground running once we get to IndieBio.
Dave and Arturo out front of IndieBio’s labs
March 2, 2015
Clara Foods gets to work at IndieBio – rigorously building out their business plan and getting in touch with customers and investors. Their science is moving along quickly thanks to their pre-work at Berkeley Biolabs, so they have samples to show.
Arturo whipping up hen-free egg whites.
May 29, 2015
Clara Foods hits the stage with New Harvest, Muufri, Pembient and Tomato Sushi to talk about the Edible Bioeconomy.
New Harvest and IndieBio co-hosted a really wonderful event on the future (or rather the emerging present) of animal products made without animals.
June 11, 2015
Arturo killing it on stage at Demo Day.
Clara and the 10 other biotech companies at IndieBio present their pitches at Demo Day. The event sold out quickly weeks before. Watch the pitches from Demo Day here!
David and Arturo at their Demo Day display.
July 9, 2015
Clara Foods announces they have completed their seed round: $1.75 million!
Within a matter of months, Clara Foods attracted $1.75 million to making animal products without animals, with the story covered in TechCrunch. Among the investors participating in the round are David Friedberg, Ali and Hadi Partovi, Scott Banister, and SOSV. Clara Foods definitely comes out of IndieBio as the most successfully invested-in company of the class – raising the money they needed even before hitting the stage at Demo Day!
Clara Foods’ hen-free egg white meringue tasted amazing.
Here’s to another great success that began with the spark that only New Harvest could create.
Written by Isha Datar July 9, 2015
An egg white is made up of around 12 key proteins. These proteins are what cause an egg white to gel when cooked, make stiff peaks when whipped, and act as a binding ingredient. These proteins are what make up the functionality of an egg white.
Today, majority of the world’s eggs are produced in high density industrial systems. Laying chickens are bred, the male chicks are culled, and the female hens are kept in small, crowded cages. Laying hens have their beaks trimmed to prevent them from injuring one another while in these confined cages. A hen will begin producing eggs at around 18-20 weeks of age, and continue to produce about one egg per day until 100-130 weeks, when the hen is slaughtered. These chickens do not become meat - they were designed for egg laying and their bones are too fragile to go through traditional meat processing.
Egg whites are usually made by hens kept in battery cages in an industrial setting. Instead, we can make the exact same egg whites by brewing them, using a culture that consumes simple sugars to make egg white proteins.
Clara Foods’ egg white proteins are made by yeast, rather than by factory farmed laying hens. Yeast is reprogrammed to produce egg white proteins by inserting the genes for egg white proteins into the yeast cells. As the yeast grows, it consumes sugar to produce the exact same egg white proteins that an ovulating hen would produce. After enough egg white proteins have been produced, the yeast and egg mixture is separated so only the egg white proteins remain.
David holding a yeast culture capable of producing egg white proteins.
Compared to producing egg whites from laying hens, producing egg whites from yeast offers many benefits.
First, there are zero animals involved in the process, which means there is no opportunity for contamination by bacteria like Salmonella and E. coli, which live in the gut and feces of animals.
Second, it is much easier to maintain a stable supply of egg proteins if they are made in yeast. In the spring of 2015, an avian flu outbreak resulted in the cull of $50 million chickens suspected of contracting the disease. This caused the price of eggs to more than double. With the crowded conditions of egg-producing factory farms, these kinds of disease outbreaks are impossible to prevent and extremely difficult to contain.
Third, because we are using a very simple organism - a yeast, which doubles every couple of hours - rather than a chicken, which must be at least 6 months before it can begin laying eggs, it should require less land and water inputs to produce hen-free egg whites. It should also produce fewer waste products and a much safer working environment for egg white producers.
Using a yeast based system to produce egg whites also allows us to do things we could never do before. For example, because we are choosing which proteins and components to produce, we can tailor the mixture of egg proteins. With slight modifications to the proportions of egg white proteins, we can create egg whites that can whip up fluffier or have better binding capabilities.
Written by Isha Datar November 4, 2015
Environmental Impact Assessment
In 2008, New Harvest funded an environmental impact assessment of cultured meat compared to conventionally produced livestock at Oxford University by Hanna Tuomisto. It was published in Environmental Science & Technology in 2011 and can be accessed here.
It was the first time there were numbers associated with the environmental benefit of cultured meat and has since been cited countless times by both researchers and the media.
The study sparked further investigation into the impacts of cultured meat production, independent of New Harvest. Hanna published further research in 2012 and 2014, and other researchers in the United States published a new life cycle assessment on cultured meat in 2015.
On August 12, 2008, Jason Matheny, New Harvest's founder and Executive Director put out a request-for-proposals on investigating the environmental impacts of cultured meat. The research was to be peer-reviewed and published in an academic journal - New Harvest would simply fund the work. New Harvest contributed $10,000 to this research.
Hanna Tuomisto at Oxford University submitted the selected proposal. She was to perform a life cycle assessment, based on a review of current research, of the impact of cultured meat production as compared to conventional meat production. The authors explored the impacts that cultured meat would have on greenhouse gas emissions, land and water use, as well as opportunity costs of land use, should it completely replace conventional meat production in all 27 member states of the European Union.
On June 17, 2011, the paper was published in the journal Environmental Science & Technology.
The paper predicted that, based on current published figures and research, complete replacement of conventional meat with cultured meat would result in an incredible 78-98% reduction in greenhouse gas emissions, 99% reduction in land use and 82-96% reduction in water use,and 45% reduction in energy use.
Comparison of primary energy input, greenhouse gas (GHG) emissions, land use, and water use of cultured meat production with conventionally produced European beef, sheep, pork and poultry per 1000 kg edible meat as a percent of the impacts of the product with the highest impact in each impact category.
In addition, the authors note that cultured meat could help reduce most environmental impacts of livestock production if the land released from livestock farming was used for providing environmental services. Cultured meat production could have potential benefits for wildlife conservation for two main reasons:
i. It reduces pressure for converting natural habitats to agricultural land
ii. It provides an alternative way of producing meat from endangered and rare species that are currently over hunted and over fished for food
However, they point out that it may also have some negative impacts on rural biodiversity due to the reduction in need for grasslands and pastures, depending upon how the land is used. In some hilly areas, livestock plays an important role in maintaining the open landscapes that are preferred over forested hills. The conversion of grasslands into forest, and arable-lands to producing energy crops for biofuel, for example, might benefit some species whilst others may suffer.
The Need for Further Research
The main criticism is that since a cultured meat production industry doesn’t yet exist, the paper is basically “a fairly complex thought experiment.” It therefore is based on a number of assumptions, such as the use of Cyanobacteria hydrolysate (an algae that can be grown in large ponds) as the nutrient and energy source for muscle cell growth, when we have not yet proven that meat tissue can even be grown using this nutrient source.
One significant problematic assumption was made in the Methodology and Data section of Hanna's paper: "As cells produce heat during the growth, additional energy inputs in heating of the reactor are not required." Many researchers have pointed out that this is an extremely unlikely scenario.
The authors do acknowledge that “as the technology for producing cultured meat in large-scale production plants is currently not well defined, there are many uncertainties about the data of the environmental impacts of cultured meat production presented in this paper.” Tuomisto and others are working towards more accurate speculative life cycle assessments. This becomes easier as cultured meat research advances and more is known about the processes involved.
Other considerations include the use of energy from renewable sources, nutrients from sources other than Cyanobacteria, and water measurements that consider green, blue, and grey water footprints.
The Importance of This Work
This initial environmental assessment paper has been cited a countless number of times by both researchers and the media.
For the first time, there were numbers associated with the environmental benefit of cultured meat over conventionally produced livestock. The paper helped to extend New Harvest's mission from one focused on animal welfare to one also focused on the environment and mitigating climate change.
The goal of this research was to expand interest and investigation on cultured meat. The media surge associated with the numbers from this study helped 1) attract more people to cultured meat as an area of research, and 2) stimulate further life cycle assessment work.
Written by Isha Datar on July 22, 2014
The 2014 research included input from Marianne Ellis, a chemical engineer at the University of Bath who focuses on bioreactor design. Marianne was involved to help correct for the assumption that heat would not be required in a cultured meat bioreactor. The paper also considered different feedstock sources of nutrients, including wheat and corn.
Comparison of environmental impacts of cultured meat with European livestock meat.
The paper compared land use and global warming potential of cultured meat to meat as other protein sources as well.
As the authors note, "We conclude that the uncertainties in the environmental impacts of cultured meat remain high", and that "more research on development of cultured meat technology is needed before more reliable estimates of the environmental impacts can be provided." We agree.
Written by Isha Datar on November 19, 2015
Carolyn S. Mattick, Amy E. Landis, Braden R. Allenby, and Nicholas J. Genovese, researchers in the United States, have published new life cycle assessment work on cultured meat in the journal Environmental Science & Technology.
Carolyn, Amy, and Brad also published an article about their research in Slate magazine. Carolyn and Brad published another piece in Issues in Science and Technology. Their work has stimulated much conversation on cultured meat's ability to minimize our ecological footprint.
We're excited to see a continued discussion, independent of New Harvest, on the impacts of cultured meat production. We hope to spark further investigation in this worthwhile area of research.
Written by Isha Datar on November 19, 2015
Mark Post's Cultured Beef
In August 2013, the first cell-cultured hamburger was cooked and tasted live on air in London, England. Professor Mark Post created the burger at the University of Maastricht. However, the majority of the muscle strands were grown in media with fetal bovine serum. In early 2015, New Harvest provided Mark’s lab with $50,000 to conduct additional research towards a completely animal-free system for growing cultured meat.
The first hamburger made entirely from cell cultured beef, made up of 20,000 muscle strands grown individually in Dr. Mark Post’s laboratory at Maastricht University. Photo by David Parry/PA Wire
The first cell cultured beef hamburger was an important milestone in cellular agriculture. It showed the world that creating cultured meat was scientifically possible - and that from here onwards, the most important innovations in culturing meat will be how to produce it in large quantities for a reasonable price. In other words, thanks to the cultured beef burger, we know producing cultured meat is not a question of scientific discovery - it’s an engineering problem.
This image from Mark Post’s lab shows how muscle tissue is routinely grown in culture at laboratory scale. The cultured beef burger was created by growing muscle strands at this scale thousands of times.
The first government funded research on cultured meat took place in the Netherlands.
The Dutch government agency SenterNovem funded cultured meat research from 2005 to 2009, in part because of conversations between New Harvest founder Jason Matheny and the Dutch Minister of Agriculture in October of 2004.
The research program was initiated by Willem Van Eelen, an 86-year-old entrepreneur who had been fascinated by cultured meat for decades and had filed a patent on the idea in 1997. Dr. Henk Haagsman, of Utrecht University, was the principal grant writer for the Dutch cultured meat project. The project was subdivided into 3 different areas: a) stem cell biology, conducted at Utrecht University by Dr. Henk Haagsman and Dr. Bernard Roelen; b) tissue engineering, conducted at Eindhoven Technical University by Dr. Carlijn Bouten; and c) culture media, conducted at the University of Amsterdam by Dr. Klaas Hellingwerf. Included in the grant application were Meester Stegeman BV (at that time part of Sara Lee foods, Peter Verstrate) and Vitro Meat BV (Willem van Eelen). During this time, New Harvest contributed $7000 to Henk Haagsman’s lab.
In 2008, part way through the project, Dr. Carlijn Bouten had to pass on her project responsibilities. Dr. Mark Post was a part-time professor at Eindhoven Technical University at the time, and took the reins of the tissue engineering part of the project by supervising Dr. Bouten's PhD students. Motivated by the potential impact of cultured meat, Mark continued research even after the grant ended.
Peter Verstrate, left and Mark Post, right, in the lab at Maastricht University.
In 2010, a large family foundation reached out to New Harvest founder Jason Matheny, asking which labs to support to advance cultured meat. Jason directed the foundation to Mark. The foundation was Google co-founder Sergey Brin’s family foundation. They worked with Mark towards a huge media event where the first cultured hamburger would be tasted, supporting the costs of the research and the event.
The cultured beef burger tasting event from August 5, 2013.
In August of 2013, the first cell cultured hamburger was cooked and tasted live on air in London, England. The event was attended by 200 journalists from around the world, the academics who worked on the Dutch Cultured Meat Project, and New Harvest.
The burger was made up of around 20,000 muscle strands grown in Mark’s laboratory. It was made with a little egg powder and breadcrumbs and a few other common burger ingredients. It was tasted by Mark, food writer Josh Schonwald and nutritional researcher Hanni Rützler. The burger was cooked by Chef Richard McGeown, of Couch’s Great House Restaurant in Cornwall.
The purpose of the cultured beef hamburger was to show the world that it was scientifically possible create an edible beef burger from cultured cells. The process of creating the burger used primarily tried and tested methods of muscle cell tissue culture.
A photo from Dr. Post’s lab by Daan Luining. 10-layer tissue culture flasks used for making a piece of hamburger. Making the entire hamburger took many more culture flasks than what is shown here.
The burger cost €250,000 to produce. It cost this much because the project took place at laboratory scale. The technicians making the burger did so by producing very small strands of beef in standard tissue culture flasks, and repeating this work several thousands of times. The price of the burger was so high because it included the salaries of skilled technicians doing very time-consuming work and consuming expensive laboratory supplies. Anon Essen, Sanne Verbrugge, and Ruud Theunissen were the laboratory technicians that did the bench work to create the first cultured beef hamburger.
A photo from Dr. Post’s lab of a technician maintaining a muscle cell culture.
One challenge in producing the burger was how to help the muscle fibers mature. Muscle fibers mature much better with contraction. Researchers in Mark’s lab found that seeding muscle cells around a cylinder of gel allowed the cells to create a fiber in the shape of a ring, which then could contract on itself.
How Mark Post’s lab created the cultured beef burger.
A second challenge was growing a large number of muscle cells. Because muscle grows only about 0.5mm in culture, the best way to mass produce the culture is by providing a lot of surface area for muscle cells to grow on.
Because cells can only grow about 0.5mm thick in culture, it is easier to grow "ground" beef. Muscle cells could be grown on beads, which offer a lot of surface area, in a bioreactor, and when the muscle cells are removed, it will already have the consistency of hamburger.
Mark Post's lab worked on some projects involving growing muscle cells on beads.
Muscle cells (nuclei stained blue with DAPI) growing on the surface of microcarrier beads (stained red with rhodamine) from Dr. Mark Post's lab at the University of Maastricht. Muscle cells need to grow attached onto a surface. Beads offer a lot of surface area, ideal for growing a lot of cells.
Another challenge in producing the cell cultured beef burger was creating an animal-free system. The status quo for culturing tissue involves the use of fetal bovine serum, an unsustainable byproduct of the livestock industry collected from fetal cows. Like other animal products, there is a lot of variation from batch to batch, an inconsistent supply, and possibility of contamination. It is the status quo because it works very well, despite little understanding on why it works. It is also relatively inexpensive.
In the creation of the cultured beef burger, Mark’s lab experimented with animal-free media for the cells to grow in. By the end of the production of the burger, the muscle strands were grown in media with zero fetal bovine serum. In early 2015, New Harvest provided Mark’s lab with $50,000 to do more research on a completely animal-free system for growing cultured meat.
Written by Isha Datar with help from Daan Luining November 3, 2015