Shojinmeat Project - Open source cellular agriculture initiative

Shojinmeat Project - Open source cellular agriculture initiative
Shojinmeat Project : Open-Source Cell-Ag Initiative  2020.01
“Shojinmeat Project” - Who we are “Democratization of cellular agriculture” Nonprofit non-corporate non-university citizen science community of DIY bio/fab enthusiasts, students, researcher, artists, writers etc. for cellular agriculture Open source cellular agriculture” by DIY bio Public communication by art and education
Ongoing projects ・DIY bio & cell culture experiments ・Workshops and micro-conferences ・Advocacy for cellular agriculture ・Media and social communication ・Art, design and creativity projects ・Visual contents production ・Fundraising / crowdfunding Creative Cells Kyoto, @CleanmeatKyoto @officialsotakan , @_okgw
Means of food production Hunting Farming Domestication Fermentation Synthesis Cell culture
“Meat”
From where? Meat is ~x40 more resource intensive Lamb：~x50, Beef：~x40, Pork：~x20, Poultry：~x7 “Meat”←animals←feed, water, land
Deforestation Public health hazard Water shortage “Meat”←animals←feed, water, land Hoekstra, Mekonnen, PNAS 2012 http://www.pnas.org/content/109/9/3232 Poore, Nemecek, Science 2018 http://science.sciencemag.org/content/360/6392/987 Ventola, NCBI 2015, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4378521/
Food vs. Feed vs. Fuel Agri- cultural resources Food Feed Fuel Poore, Nemecek, Science 2018 http://science.sciencemag.org/content/360/6392/987
Anticipated alternative proteins *As demand for protein grows, existing meat cannot sustainably serve for all. MeatSoy etc. Dairy MeatDairySoy etc. New alternative protein Plant-based Tofu Algae Insects Biosynthetic Cell-based New protein source “Meat & dairy produced in new ways” Plant-based meat & dairy equivalent Now Future
Cellular Agriculture Agricultural products by cell culture Medicine technology, Agriculture application
Cell-based meat (cultured meat) Muscle cells Bioreactor Culture medium Processing
Energy conversion efficiency < 0.1% Prospectively, Microalgae: 4~11%? “Artificial leaf”: 10%+? https://aiche.onlinelibrary.wiley.com/doi/abs/10.100 2/btpr.2941 Y.Okamoto et al. Biotech. Prog. 2019 ~4% ~35% 50~90% Unregulated Shepon, Eschel et al, IOP Science 2016 http://iopscience.iop.org/article/10.1088/17 48-9326/11/10/105002/meta
Ongoing cellular agriculture projects Wild Type Fish without catch IntegriCulture Foie gras without ducks Shiok Meats Shrimp without pond or catch The products are not “imitations” - they are (or try to be) molecularly the same! Perfect Day Milk without cows Clara Foods Egg white without chicken Ginkgo Bioworks Vanilla, scent, various ingredients
Research track record 1 1997 Goldfish meat @NASA Appearance in sci-fi - Concept known since 19th century 2004 New Harvest founded Jason Matheny contacts NASA staff 2005 Netherlands funds €2M 2007 In vitro meat consortium funding discontinued funds 2000 Works by Oron Catts @Harvard Lead by Willem van Eelen (deceased) Patent filed in 1997
Research track record 2 2012 Sergey Brin from Google contacts former member of In vitro meat consortium 2013 Demonstration by Prof. Post 2014 Shojinmeat Project 2015 Memphis Meats 2015 “Cellular agriculture” - term coined 2013 New Harvest invests in cell-ag startups (Clara Foods, Perfect Day) 2016 SuperMeat 2017 Finless Foods €280k burger
The positive impact 學經藝 Science & Technology ・Technological hurdles? ・Medical applications? Politics & Economics ・Shift in food market? ・Industry landscape? Arts and Culture ・Religious views? ・Social norms to change? Hanna Tuomisto　2011
The technical problem $325,000 Cell-based burger, 200g
Technological milestones 1. Inexpensive media 2. Scaling & automation CapEx &  Staffing  Culture medium $200k+/kg Conventional method $2 Conventional meat price & quality parity? 3. Added value and consumer acceptance Technological goals 1. Food grade culture medium 2. Scaled culture plant design 3. Tissue engineering for flavor and texture
Gospodarowicz D and Moran JS, 1976, Annu Rev Biochem Eagle H, 1959, Science ＝ Culture medium Expensive for what’s actually in (as they are made by the bio-pharma grade) Expensive and insecure supply, and serum risks infectious diseases - viruses, “mad cow” etc. Made by fermentation of recombinants, but regulatory approval and extraction are expensive. Basal medium Foetal bovine serum Signal compounds Sugar, Amino acids, Vitamins, Minerals  $20/L Albumin, Buffer, Insulin, Transferin  $900/L Growth factors Survival factors  $450/mg Bottlenecks in culture medium
Standard DMEM(FBS10%) 500ml DMEM 450ml $10 Serum(FBS) 50ml $45 Non essential amino acid $1.30 LIF　 10 U/ml $2.00　etc・・・ Medium for 1~2g of cells： $58.30～ $5000+ for 100g Cost of culture media for “easy” cells
Myoblast culture by Essential 8® defined serum An analysis of culture medium costs and production volumes for cell-based meat, Liz Specht, Good Food Institute (2018) https://www.gfi.org/files/sci-tech/clean-meat-producti on-volume-and-medium-cost.pdf Basal medium DMEM components The E8 medium based on TeSR medium does not contain albumin and other animal-based materials, and its composition is publicly known. Its price is around $380/L. Serum ITS, AA2P Signal compounds FGF2, TGFβ $1.60 Bulk-purchase price $5.44 $181.36 AA2P：Vitamin C derivative ITS：Insulin, transferin, Na2Se 500ml culture medium 100g of cells $188.40 $15,000~ Myoblast requires FGF2 and TGFβ peptides to culture. Besides peptides, some fatty acids and cholesterols also act as signaling compounds.
Hepatocyte (liver cells) and iPSC’s, 100g Basal medium Serum Signal compounds Hepatocyte iPSC(medical) DMEM 450ml $10 FBS 50ml $45 Additional amino acids $1.30 HGF 20µg ~$700 EGF 10µg ~$6.40 ~$760 ~$90,000 ~$3000 $300,000+? Specialised medium, ~$1000/L Often not required Multiple GF’s i.e. “bFGF” consumed for each steps Block et al., 1996 https://www.ncbi.nlm.nih.gov/pubmed/8601590 iPS cornea costed ~$1M https://biz-journal.jp/2015/06/post_10151.html 500ml culture medium 100g of cells
Cost of goods analysis [1]GFI’s white paper https://www.gfi.org/files/sci-tech /clean-meat-production-volume- and-medium-cost.pdf Basal medium Serum & Signal compounds Material[2] Price Purpose Why expensive Possible actions NaCl $56 Medium Used a lot Recycle medium Glucose $50 Cell mass Used a lot Less expensive procurement channel Amino acids $395 Cell mass Used a lot Less expensive procurement channel HEPES $3933 Medium High unit price Use an alternative, recycle medium AA2P $10,035 Medium High unit price Use an alternative, recycle medium Cost of goods analysis of E8 medium per 20,000L(3.5t of meat) - bulk procurement assumed[1] Insulin $131,920 Serum High unit price Use an alternative Transferin $85,600 Serum High unit price Use an alternative FGF2 $4,010,000 GF Extreme price Use an alternative TGFβ $3,236,000 GF Extreme price Use an alternative [2] NaCl, HEPES and AA2P remains in medium while glucose and amino acids are consumed or converted into cell mass. [3] Serum and signal compounds are expensive, but can not be recycled as they are consumed.
Driving down the cost : Strategy A Efficiency Stability Unit price Alternative Actions taken References Expert interview on less expensive growth factors in Good Food Institute’s grant program: Dr. Peter Stogios, Senior Research Associate, University of Toronto, Canada (2019): https://www.gfi.org/dr-peter-stogios-grow th-factor-research GFI competitive grant program https://www.gfi.org/2019-competitive-res earch-grant-program Use alternatives such as fatty acids and egg yolk components Design the growth factor molecules to increase the efficiency and reduce the required quantity Use of larger bioreactors to grow recombinants, use of plant-based GF’s, cheaper extraction method etc. Design the growth factor to give higher structural and thermal stability to withstand less expensive extraction methods Serum & Signal compounds
Cost of goods (ingredients) analysis Medium Meat $41/L $100/lb $15/L $36.6/lb $4.7/L $11.5/lb $3.7/L $9.0/lb $0.77/L $2.2/lb Scenario A~E A: All GF’s down to 1/10 in cost B: FGF2 & TGFβ at insulin price C: A&B D: All GF’s at $4/g E: Basal medium at $0.23/L “Strategy A” assumed Analysis by the Good Food Institute (2019) https://www.gfi.org/files/sci-tech/clean-meat-production-volume-and-medium-cost.pdf
Cocultured hepatocyte produces serum and signal compounds without a need for their external addition. PCT/JP2016/067599 JP Pat.6111510 Integriculture Inc. / Dr. Ikko 2016 Strategy B : Large-scale coculture Feeder bioreactor system by IntegriCulture (Japan)Quadruple co-culture by Aleph Farms (Israel) https://www.aleph-farms.com/#RD Basal medium Serum & Signal compounds 4 cell species mutually stimulate cell proliferation, i.e. fat cells enhances muscle cell proliferation. Basal mediumSerum & Signal compounds Myoblast Hepatocyte
Demonstration of coculture Significant hepatocyte growth without added growth factors (HGF)Control (0%) 10% conditioned medium 25% conditioned medium 50% conditioned medium Countofcellsofallsizes, relativetothecontrol group mouse placental cells dishes with Day-12 foetal liver cells in FBS 10% medium 7 Days Transfer culture medium Dr. Ikko 2016
Cell-based chicken liver demonstration cell-based chicken liver paste http://www.nicovideo.jp/watch/sm32032224 Grow cells Fat-load cells Assemble cells Dr. Ikko 2016
Low-cost Liver cell culture Figure by Integriculture Inc. DMEM　 450ml　 $10 Non essential amino acid　 $13.0 FBS 50ml $45 HGF 40ng/ml $700 (20µg) EGF 20ng/ml $6.40(10µg) $760~ Liver cell aggregate on collagen scaffold- HGF/EGF obtained from cocultured cells DMEM　 450ml　 $10 50ml $1.70 ~$12 yeast extract, an FBS alternative Yeast extract
“Food grade DMEM” Culture medium, as inexpensive as bottled beverages? DMEM　 450ml　 $10 50ml $1.70 ~$12 ”DMEM” 450ml　 $0.09 50ml $0.01 $0.10 Sugar Amino acids Vitamins Minerals Basal medium = Yeast extract Yeast extract Use of yeast extract marketed as food, not laboratory reagent or its equivalent
“Food grade” demo - Sports drink culture media 90% DAKARA 80% 70% 60% 0% (DMEM only) 50% DaysCelldivisions Proliferation of mouse L6 in DMEM/GreenDakara 10% FBS #pH and osmolality of Dakara adjusted by NaHCO3(s) and 2M NaOH Fluid name osmolality pH DMEM(-, hi glu) 345 7.4 Pocari 338 3.4 Aquarius 291 3.37 Amino-Value 4000 289 3.63 AminoVital Gold 186 3.33 Vitamin Water 302 3.3 Green DAKARA 322 3.28 Amiiru Water 249 3.4 Mamoru Chikara 546 3.58
DIY-DMEM (home made medium) http://animescience.net/?p=3647 http://www.nicovideo.jp/watch/sm33104702 Glucose, amino acids, vitamin B’s, salts Mix and filter to prepare Chicken foetus heart cells primary culture Protein supplement Vitamin pills Glucose Salts
Basal medium turning commodity product Individual ingredients Digested yeast residue? Algae? Artificial photosynthesis? Bulk purchase may partly reduce costs Mixture of 20 amino acids, along with undigested dipeptides and tripeptides - impurities are acceptable as long as cells can be cultured, even with lower efficiency - the cheapest ”Minimum grade medium”? Digest Production at a Mega-ton scale？ Alage Artificial leaves Power Hydrogen bacteria Electrolysis Minimum grade medium 2020 Bridging methods Basal medium as a commodity
Mainstream demand for cell-based meat requires large supply of inexpensive amino acids. Novel raw materials and production process (the incumbent is fermentation) must be developed. Rice 720Mt Wheat, Barley 830Mt Corn 880Mt Cassava, Tubers　 630Mt Soy 260Mt Sugar beet Canes, 2.1Bt Oil Palms Starch※1 40Mt Sugar 170Mt Meats　300Mt (Beef 60Mt, Pork 110Mt, Poultry 80Mt※3 ) Bioethanol※2 100BL Amino acids※4 6Mt Oil 40Mt Food (Carbs) Cell-based meat Plant-based protein sources Process residue Food (Fats) Food (Carbs) Figures are as of 2011, Geographic Annals 2014 (Ninomiya Books)　※1 https://www.alic.go.jp/joho-d/joho08_000573.html 　　　※2 Monthly Report, MAFF Japan March 2015, http://www.maff.go.jp/j/zyukyu/jki/j_rep/monthly/201503/201503.html 　　※3　USDA「World Markets and Trade」　　　※4 Ajinomoto Co. Ltd.   Food (Protein) Food (Protein) Source: Mr. Akito Chinen 2nd Cell-Ag Conf. in Japan Sources of amino acids for cell-based meat
“Matter cycle” of cell-based meat Convert to culture media Large-scale cell cultureWaste fluid treatment Algal production Algae sewage Fertilizer
“Is it tasty?” - Tissue engineering to add value Sausage/burger Proven ※although expensive Low cost large scale cell culture Sheet meat / “bacon” Cell scaffold Muscle/fat coculture Steak / tissue Tissue morphogenesis Vascularization Meat texture Regenerative medicine Where we are
Tissue engineering, regenerative medicine and cellular agriculture Cell culture (human) Cell culture (animals) Distribution Distribution Regenerative medicine Cellular agriculture Med. Ag. Procu- rement Culture medium Cell-ag and Regen. Medicine share the same technology. Main differences are in purity, traceability and regulations
Building “meat” : Method 1 “Opportunities for applying bio- medical production and manufacturing methods to the development of the clean meat industry” https://www.sciencedirect.com/science/articl e/pii/S1369703X1830024XCulture medium and starter cells Myoblast culture Myogenesis, Cell organization Grow cells Build tissue by cellular scaffoldProcurement
3D culture by cellular scaffolds Cellular scaffold has large surface area, which improves the efficiency of cell culture. Sponge collagen scaffolds Liver cells on scaffolds
Other functions of cellular scaffolds Edible scaffolds i.e. collagen, chitosan, chitin, arginate, cellu- lose, polysaccharides Simulate fibre and meat texture Moulds shape in mm or bigger scales Aleph Farms (2019) J.R. Gershlak et al., Biomaterials vol.125, pp13-22(2017) https://www.sciencedirect.com/science/articl e/pii/S0142961217300856?via%3Dihub
Building “meat” : Method 2 ”In Vitro Engineering of Vascularized Tissue Surrogates” https://www.nature.com/articles/srep01316 Culture medium and starter cells Procurement Cell growth, vascularization Tissue growth ※Further down the development timeline
Tissue engineering to and beyond thick steak Cells on scaffold around 2020 Cell aggregate 2013 Cultured tissue 2026? Designer meat? 2030? Burger, sausage Bacon, meat chips Thick steak Is this even ”meat”!? Algae-myoblast co-culture
Scaling as it is... How cultured burger was made in 2013 ⇒$260,000
Conventional “scaled” cell culture Needs much more scale
The process needs a fundamental re-design Labs, Hospitals Brewery, Petrochemical complex Culturing of cells has been optimised for laboratory scale ⇒€250k per burger Culturing of cells becomes industrial scale ⇒Production at $2/kg
Dr. Marianne Ellis, 2017 Implementation of scaled production methods ⇒”Chemical engineering”, “Plant Engineering”Integration and systemization “Cell culture processes suitable for scaling”
How is temperature controlled? Mixing method? Pipeline diameter & flow rate? Sterilization method & frequency? How are filters cleaned? Plant engineering - what exactly? Speculative fish meat culture plant
Production strategy and estimated cost A：”Batch” B：”Fed batch” C：”Perfusion” A：Produce (culture) cells for ~40 days, harvest once full B：Produce by feeding the batch until waste metabolites build up C：Produce while feeding and recycling the culture medium ←Estimated medium cost of cell- based meat based on method C- $1.37/kg　(GFI 2019) Lipid Nanocarriers for Drug Targeting https://www.sciencedirect.com/book/97801281 36874/lipid-nanocarriers-for-drug-targeting Reference https://www.gfi.org/files/sci-tech /clean-meat-production-volume- and-medium-cost.pdf
Comprehensive cost and footprint assessment ・Resource requirement from ‘cradle to the grave’ Life cycle assessment (LCA) https://rationalwiki.org/wiki/File:LCA.PNG
LCA uncertainties Current quantitative LCA estimates depend on unestablished production processes and uncertainties may exceed 50%. LCA is expected to be a key point of discussion in international standardization of production processes. 2014 estimate included steam sterilization of bioreactors. Hanna Tuomisto　2011 Hanna Tuomisto　2014
Animal co-products and “secondary LCA” Co-product alternatives By synthetic materials and products[1] Gelatin production by cell-ag[2] Source of energy Without decarbonization of the power source, GHG reduction may be limited.[3] How is land freed from animal agriculture be used? (Secondary LCA) The environmental footprint taking into account the alternative land use[4] [1]https://www.atkearney.com/retail/article/?/a/how-will-cultured-meat-and-meat-alt ernatives-disrupt-the-agricultural-and-food-industry (AT Kearney) [2] https://geltor.com/ Geltor Inc. markets cellular agriculture gelatin [3] https://www.frontiersin.org/articles/10.3389/fsufs.2019.00005/full Footprint of cultured meat if coal power remains for the next 1000 years [4]https://drive.google.com/drive/u/0/folders/0B1yPiUX43toZY2sxUUtVRjlZTDA Tuomisto et al. 2017, Int’l Sym. Cultured Meat, (unpublished) Other products include manure, gelatin etc.
“Protein problem” in different stages Is there enough protein? Is it a secure source? Is it sustainable? Many rely on imports while overfishing continues ( Economic and Social Research Institute, Dublin, Ireland, 2010 https://www.esri.ie/pubs/WP340.pdf) “Public expenses due to meat is set to reach $1.6T by 2050” Farm Animal Investment Risk and Return Initiative Poor conturies
“Impending food crisis”? ”Boiling frog" ・Food prices rise incrementally over decades ・Lower living standards, more frequent civil unrest
How cellular agriculture is a solution 1 2 Reduce protein consumption. Change the way we eat. Find sustainable protein source. Preserve culinary culture. Meat supply chain Externalities “Wicked problem” of climate change, poverty and local ecological losses
Global protein market outlook Population growth and emerging economies: $2T market cap. by 2030? Global meat demand 1980-2030 Rabobank (2011)■Lamb ■Poultry ■Pork ■Beef Demand,10milliontons
By Olivia Fox Cabane, http://newprotein.org ※Also describes investors (VC’s, corporates) 　 IP info: https://www.culturedabundance.com/about Startups list: https://futurefoodshow.com/list-of-cultured-meat-companies/ Japan $50B Seafood $250B Global meat $1.9T Global beef $0.7T Market size along the value chain of meat (in billion $) Source: AT Kearney Cellular agriculture startups and market size
“Brewed milk”, ice cream, 2019 https://www.perfectdayfoods.com/ https://thespoon.tech/perfect-day-launches-ice-cream -made-from-cow-free-milk-and-we-tried-it/ Launch dates of products Product Launch dates?（MosaMeat https://www.mosameat.com/faq ） “We are aiming for a first market introduction in the next few years. It is very difficult to commit to a particular timeframe because there are still some scientific unknowns and factors outside our control (such as the regulatory process). The first introduction will likely be small-scale. Several years beyond that, we aim to be widely available in restaurants and supermarkets.” Cosmetics, foie gras etc. 2020~ https://integriculture.jp https://www.slideshare.net/Yuki Hanyu/ss-166477453 Shrimp, 2021~ https://shiokmeats.com https://www.undercurrentnews.co m/2019/09/12/shiok-meats-says-l ab-grown-shrimp-meat-will-be-on -market-in-2-years/ Collagen for cosmetics, 2019 https://geltor.com/ https://techcrunch.com/2019/11/01/ after-signing-a-big-food-additive-de al-cell-based-protein-company-gelt or-is-looking-for-at-least-50m/ Beef, fat tissue, 2022～ https://www.mosameat.com/ https://www.labiotech.eu/food/mosa-me at-lab-grown-meat-fundraising/ Beef steak, 2021~ https://www.aleph-farms.com/ https://www.euronews.com/2019/07/17/israel i-company-aims-to-have-world-s-first-lab-gro wn-steak-on-market-by-2021 Duck, poultry, beef etc. 2021~ https://www.memphismeats.com/ https://www.livekindly.co/memphis-meats-brin g-clean-duck-chicken-meat-stores-2021/ Blue fin tuna etc., 2021～ https://finlessfoods.com/ https://www.foodnavigator-asia.com/Article/2018/0 6/25/Clean-fish-Finless-Foods-reports-success-in-s lashing-production-costs-for-lab-grown-products Bioreactor systems, 2022~ https://www.future-meat.com/ https://techcrunch.com/2019/10/10/lab-gro wn-meat-could-be-on-store-shelves-by-20 22-thanks-to-future-meat-technologies/
Cell-ag market predictions (By A.T.Kearney) ・⅓ becomes meat alternatives by 2030. ・Plant-based meat in 2035 reaches maturity. ・CAGR 40% is expected for cell-based meat. ・The shift will also affect surrounding sectors i.e. retail and distribution. Source reference https://www.kearney.com/documents/2 0152/2795757/How+Will+Cultured+Me at+and+Meat+Alternatives+Disrupt+th e+Agricultural+and+Food+Industry.pdf/ 06ec385b-63a1-71d2-c081-51c07ab88 ad1?t=1559860712714
The road ahead in protein transformation More information: https://www.slideshare.net/2co/r oadmap-to-the-cellular-agricultur e-era Improving technology reduces the amount of capital for market entry. Individuals enter and technology democratizes. Hype crash and cycle Reaches price parity with animal meat Conversion by regions and product categories Vertical integration with large plants 2020～ 2030～Animal meat Cell-based meat Price Time Foie gras Beef Chicken Importer Half-producer Producer
“Adjacent industries” Entry from nearby fields Beverage companies Food companies Medical supply manufacturer Plant engineering firms Effect to nearby fields Functional & pharmaceuti cal foods Food branding Biomanu facturing Regen. medicine Indoor farms Cell-based meat industry mind map Good Food Institute (2016)
Non-profits USA IsraelUSA Japan USA ・Plant-based and cell-based meat advocacy group founded by New Crop Capital that promotes cell-ag from the industry side. ・GFI has branches in global locations to act on governments. ・Headquartered in Washington DC https://www.gfi.org/ ・Funded by crowdfunding ・Promotes open-source DIY cellular agriculture ・Spun off IntegriCulture Inc. and CAIC http://shojinmeat.com Founded by university students, aims to connect academia and industry https://www.cellag.org/ ・Originally animal welfare centered group ・Spun off SuperMeat and FM Technologies https://www.futuremeat.org/ etc. ・Donor-funded 501(c)(3) that coined the term “cellular agriculture” in 2015 ・Supports its academic research and one of the key advisors to FDA https://www.new-harvest.org Spin-off Cell-Ag Inst. of the Commons Academic and policy advisory and public communication ※Still in preparation on 2020.01 https://cellagri.org Lobbying etc. Grants
Notable conferences Alternative protein and cell-ag sessions are also common in ”Food Tech” events, i.e. Smart Kitchen Summit (Seattle) and Agri-Food Innovation Week (Singapore) ~Sept. in San Francisco by GFI, More industry oriented, on both plant-based and cell-based meat https://goodfoodconference.com ~Oct. in Maastricht, Netherlands, An academic conference https://www.culturedmeatconference.com/ ~July in Boston by New Harvest, Academics and biohackers https://www.new-harvest.org/new_harvest_2019 ~Nov. in San Francisco, cell-ag specialized industry event https://2019.cmsymp.com/ ~Feb. in San Francisco, Industry meeting by Hanson Wade(UK) https://industrializingcellbasedmeats.com
Academic researches Supports New Harvest Demonstrated cell- based meat in 2013 Japan Science and Technology Agency “JST-Mirai” program (government grant), ~$20M for FY2018-2023 https://www.jst.go.jp/mirai/jp/uploads/application-guideline -h30-en.pdf While startups raise large amounts, Dolgin points out on a Nature article the relative lack of basic research in cell-ag https://www.nature.com/articles/d41586-019-00373-w $3M grant program (2018, 2019) https://www.gfi.org/researchgrants https://www.gfi.org/blog-cultivated-meat-industry-needs 2019~ entrants from other fields https://www.nature.com/articles /s41538-019-0054-8#Abs1 Past (2015~) research projects https://www.new-harvest.org/past_research_projects Latest research grants https://www.new-harvest.org/current_research_projects
Government actions on cellular agriculture Acknowledging “cell-based meat” as “meat”, USDA and FDA agree on March 2019 to jointly regulate cell-based meat, detailed regulatory frameworks to follow https://www.fda.gov/news-events/press-announcements/usda-and-fda-announce- formal-agreement-regulate-cell-cultured-food-products-cell-lines-livestock-and Cell-ag startups launch an industry coalition “AMPS Innovation” https://ampsinnovation.org/ (EU)2015/2283 sets cell-ag products as novel food and applications are handled by the European Commission & EFSA. https://eur-lex.europa.eu/legal-content/en/TXT/?uri=CELEX%3A32015R2283 Funds $3M to Meatable in 2019 Cultured pork demonstration in Nanjing Agricultural University http://news.njau.edu.cn/2019/1123/c39a104311/page.htm Promised to invest maximum $300M on Israeli alternative protein startups and some already realized. Government fund invests in a cell-ag startup. Government officials and policy makers submit policy advices and launches rulemaking activities. http://www.shugiin.go.jp/internet/itdb_shitsumon.nsf/html/shitsumon/a200106.htm Launches FoodInnovate program to improve the nation’s low (10%) self sufficiency rate - actual players include A*Star, EDB, AVA and SFA (2018) https://www.straitstimes.com/singapore/health/new-government-strateg y-will-drive-food-innovation https://www.foodnavigator-asia.com/Article/2020/01/22/Next-gen-produ cts-Safety-first-as-Singapore-delves-into-regulatory-framework-for-cell- based-meat Government funds $640K to two research institutions in Hyderabad on cell-based lamb meat https://indianexpress.com/article/india/centre-to-fund-res earch-on-cell-based-meat-by-two-institutes-5695154/
Inclusive rulemaking on cellular agriculture Food companies Farmers Cell-ag startups Academics Policy makers General public Additional Potential stakeholders (New Harvest / GFI equivalent in Japan) Wishlist Wishlist for policy makers & regulators, with representations of included parties Ideally, No one left behind in “cell-ag revolution”
Who will regulate meat? (And what is “meat”?) Good Food Institute, Tofurky and a few alternative protein startups filed a court case against State of Missouri https://www.nytimes.com/2018/08/28/us/missouri-meat-law-tofurky.html Lobbies and petitions not to call plant-based and cell-based meat as “meat” State of Missouri passes a Food Labeling law in favor, 25 states follow(end. 2019) USDA USDA/FDA[1] agreement 2019.03 FDA FDA USDA, as it is “meat” Discrepancy [1]While most food is under the jurisdiction of FDA, “meat and poultry” are under USDA. Cell extraction Cell culture Tissue harvest Consumer products Cell source
Nomenclature ”Cultivated meat” (used to claim “clean meat”) https://www.gfi.org/how-we-talk-about-meat-grown-without-animals cultured meat, also acknowledges “cell-based meat”) Industry players agree on the term Cell-based meat and coalition 純粋培養肉(純肉)、細胞(培養)肉・How do general consumers perceive？・Is it a neutral name? ・Does it comply with food labeling law? Good Food Institute(2017) （Japanese）
Regulations and cellular agriculture products A food product must be manufactured from food and (approved) food additives under proper process control. A “food” must have history of consumption (⇔ “Novel food”). There is a list for (FDA, in case of USA) approved food additives. cell-based meat Growth factors Could have a GRAS status Requires safety tests and approval if used. Sugar Amino acids Vitamins Minerals etc. (Food & additives) Basal media Basal media Process control: HACCP & GMP Could use sports drink
Where are the risks? BSE prions? Cells mix-ups? Viral contamination? Is it GMO？ Are growth factors safe? What’s in the medium? Bacterial contamination? Cancerous cells? Safety of cell metabolites? Will it be labelled? Unexpected health risk from “too clean meat”? Is meat healthy?
細胞肉の食品安全衛生：対策例や争点 Use of primary cells or cell line cells? Viral DNA can be monitored real-time in near future? Can be avoided by cell acquisition from prion-free parts? Use of cell sorter may separate by each cell types Existing food regulations can evaluate the food safety of growth factors? Use of methods that does not require external addition of growth factors? Use of known medium? Prompt reporting of any changes made to medium? https://www.sigmaaldrich.com/life-science/cell-culture/learning-center/media-formulations/dme.html Would require contamination detection mechanism for quality assurance “Substantial equivalence” - how do the amount of abnormal or cancerous cells compare to existing meat? How should it be labeled? “Beef(cell-based)” ? Long-term safety - how do cell-based meat differ from conventional? Is meat consumption healthy - regardless of traditional or cell-based?
Socio-cultural implications Would vegetarians eat? Is it Halal? Goes with Buddhist “nonviolence”? Animal welfare? Consumer acceptance? At the end, is it tasty?
Regional differences in agenda A. Animals, religion B. Food security C. Food safety D. Env. footprint EU/US：　A~D > C > B E.Asia：　B~C > D > A
Western vegetarianism ・Vegetarians ・Pescetarians ・Vegans are the most common 3~10% of the population
Practical ethics: “Utilitarianism” “Decision should be made to maximize utility” Utility = happiness x number of sentient beings “Only 1 death is better than 5”...? May be so in very short term, but... If “killing for public good” becomes the norm, no one is there to stop dictators → Is the utility maximized, in the long run? Utilitarian decision making is strongly dependent on subject and timeline“Runaway trolley problem” As it is, 5 will die. If you switch, only 1 will. Will you do it?
Animal welfare as deductions from utilitarianism ◆Animals are capable of feeling happiness and thus sentient. ◆All sentient beings count. ◆”Species” as defined by biology is irrelevant in ethical decisions. ⇒From a utilitarian point-of-view, “making sentient beings suffer is unethical”
Acknowledges health and environment, but mainly animal welfare and ethical “Animal welfare” based on utilitarianism Reason for being a vegetarian Vegetarian Society
East Asian vegetarianism ”Religion”←Not utilitarianism or other ethics Ethical value of clean meat described by utilitarianism don’t directly translate into East Asian religious (i.e. Buddhist) importance.
“Shojin ryori” Buddhist cuisine Cuisine for for zen practitioners Common name for “Buddhist cuisine” All aspects must serve the purpose of zen. -NO FOOD WASTE -Use local produce to avoid food waste -No cruelty (avoid fish and meat) -Avoid ingredients that stimulate desires i.e. onions -All donated food (incl. meat) must be consumed -Cooking is also a part of zen practice ※contested Known as “zhai cai” (齋菜) in Chinese
Would “murderless meat” help Zen? ”Would cell-based meat serve the purpose of Zen?” ・Overcoming personal desires is a major theme in zen ・”Desire” includes meat apetite ・”Fake meat” is a compromise, but compromise is permissible ・Cell-based meat fits in the same category as tofu - meat imitations. Cell-based meat being “murderless” does not make it Zen - there are multiple more important criteria. “Shojin” means “devotion to the righteous path” Shojinmeat Project will stay committed to the path that obsoletes unsustainable meat
Meat in historical Japan Before 7th century： Eating meat was common. People just had to eat whatever was in hand. 675c. Imperial decree of “No Killing (of animals)” To direct labour force to rice production and put a stop to local animal-sacrifice rituals & reinforce imperial authority ※Newly arrived Buddhism was used as justification Meat avoidance continues till 19th c. and commoners only started eating meat around 1900 c.
・Totals half billion? Region-specific ・More common among upper castes ・Some upper caste members fund cultured meat research ・Hinduism doesn’t explicitly forbid meat but adherents choose to avoid meat. Mr. Modi (Indian PM as of 2017) is a vegetarian. Vegetarianism in India
Is it Halal? “The halal cultured meat can be obtained if the stem cell is extracted from a (Halal) slaughtered animal, and no blood or serum is used in the process.” J Relig Health. 2018 Dec;57(6):2193-2206. https://link.springer.com/article/10.1007%2Fs10943-017-0403-3 “As long as the cells used are not from pigs, dogs or other animals banned under the halal laws, the meat would be vegetative and "similar to yogurt and fermented pickles." Abdul Qahir Qamar, The International Islamic Fiqh Academy, Jeddha, Saudi Arabia https://gulfnews.com/going-out/restaurants/is-shmeat-the-answer-in-vitro-meat-cou ld-be-the-future-of-food-1.1176127 “Work in progress, and being settled” Halal authentication by JAKIM (backed by the Malaysian government) is widely acknowledged. “It is Halal” may become the single decisive purchase reason in Islamic sphere.
Potential future shift in ethical landscape Then what if on 2040, meat alternatives rise to 30% market share and ever more people stop consuming conventional meat? ・Uncontrollable ”hate campaign” against traditional farmers? ・Trade ban of conventional meat due to animal cruelty? Why are animal experimentation, Japanese whaling and Chinese cat/ dog consumption is problematised far more than factory farming? ⇒Because they are “remote things” for the protesters.
Ethical issues due to technological immaturity (Transient) issues upon R&D：・FBS production is not cruelty-free “Unavoidable” issue：・Extraction of cells Likely to be solved in the future May pose an issue during R&D Can bovine foetus feel pain? It may still inflict some pain. Will animals still be chained? Genetic selection of animals for the sake of “tasty” - is it eugenics?
Ethical issues of “captive carnivore” Wild animals exist outside human control and no ethical questions are raised against humans. However, captive carnivores as pets and zoo animals are under human control → ethical questions ?? Is it ethical to let carnivores to prey on others??
Future court case: “Patent or Life (of animals)” ・Can rich countries with cell-based meat production technology blame (poor) emerging countries for animal abuse? ・Court cases over “economic incentives (patent) vs. animal suffering” Case study: Generic HIV drug lawsuit: An Indian pharmaceutical company allegedly infringed retroviral drug patent to manufacture generic HIV drugs, because the original drugs by Western pharmaceutical companies were too expensive for people in poor African countries. After high-profile court-martials, the Indian company won the case on humanitarian basis.
Consumer acceptance Chris Bryant (2017) 3rd Intl’ Conf. for Cultured Meat Large variations exist between different marketing research attempts and speculations, mainly due to the lack of actual clean meat products on market. ←Factors consumers weigh Consumer acceptance research result→
What would a corporate monopoly do? Technological details were concealed as “trade secret”, drawing widespread accusation and allegations of “technology for corporate profit than social good” Science may have proved the safety of GMO, but failed to convince the public to feel safe. Cell-cultured meat: Lessons from GMO adoption and resistance (Review paper) https://www.sciencedirect.com/scienc e/article/abs/pii/S0195666319304829 ?via%3Dihub
Cellular agriculture ecosystem Advocacy, Academic research with universities DIY bio, speculative art projects, “avant-garde” advocacy Sponsor Research and project grants Commercialization, Production technology development Individual biohackers in communities such as
Shojinmeat Project as citizen science Citizen Science Non-Profit Advocacy, Academic research with universities DIY bio, speculative art projects, “avant-garde” advocacy Sponsor Research and project grants Spin-off Startup Commercialization, Production technology development
Cellular agriculture with initiatives on citizens How to make ⇒ Open How to scale ⇒ Proprietary Numberindemand Degree of personalization Product dev. Citizen Science domain Business domain and other businesses to come Transparency in technology
Citizens are the decision makers Academia hints the way Citizens act and set the direction Businesses scale and deliver Wilsdon, James and Willis, Rebecca, why public engagement needs to move upstream (2004), Demos, London.
“Growing meat at home” =DIY bio methodology= Konjac cell scaffold Cells from fertilized eggs Egg white as antimycotic Egg yolk as FBS ⇒DIY cell culture in kitchen High schooler on DIY cultured meat experiment, TV news Airtight box CO2 source humidity source dish DIY cell culture manuals (mostly translated) https://drive.google.com/drive/folders/13TsVy 0JrSxFl4ZoYe6D_uPfDYr3d2P5e
Development of DIY bio equipments DIY incubator Temperature can be set to ~40℃ by reducing the AC input voltage of a towel warmer from ~100V to 30V. The blueprint of DIY incubator is on GitHub. Both are at ~$100. https://github.com/sotakan/RPi-Incubator by @okgw_ Household fan centrifuge (~100G at ~1000rpm) Egg white antimycotic and DIY clean bench http://interestor2012.hatenablog.com/ entry/2015/01/05/202524 (@earthlyworld
Development of DIY cell culture protocols Materials and methods are uploaded on blogs, video sharing sites, GitHub etc. for other participants to confirm reproducibility. Creative Cells Kyoto reproduces sports drink culture medium experiment https://cleanmeat-kyoto.hatenablog.com/ http://zacmayu.hatenablog.com/ http://www.nhk.or.jp/gendai/articles/4149/ Ms. Sugisaki - consultant by the day, biohacker at home (aired on TV)
Demonstration video online http://www.nicovideo.jp/watch/sm30403849 http://www.nicovideo.jp/watch/sm30099092 http://www.nicovideo.jp/watch/sm31104033 VR video：https://www.youtube.com/watch?v=YdELM3xjmr0
Development of home and school cell culture kit Pre-survey to high school teachers School cell culture kit should be at <$300 DIY cell culture hardwares and protocols deployed in a class Less than $100 for a class of 20 -At what budget? -Cell culture in school classes? (9 respondents) ◆<$300 ◆<$500 ◆<$1000 ◆<$3000 ◆<$5000 ◆Yes ◆No ◆ ◆ ◆ Yes, as an extracurricular activity Yes, if there is time Depends on budget @thesow41 (towel warmer) (DIY protocols)
DIY cell culture and future Other cell types? Coculture system, better DIY serum? Cell scaffold, DIY tissue engineering ….etc. What’s next? DIY cell culture experiments Materials and methods on blogs and online videos Cell culture kit  Results are shared, other participants also reproduce results Cell culture protocols  Cell passage protocols, cell library
Why stop at meat? DIY tissue engineering? Tastier than meat DIY kidney! DIY differentiation & morphogenesis to make heart Is this even “meat”!? “Green meat” algae- meat composite food https://www.ncbi.nlm.nih.gov/pmc/art icles/PMC5282507/ ・Bioreactors improve over time, enabling cell, tissue & organ culture ・DIY tissue engineering is ubiquitous, innovative prototypes everywhere
Local farmers design their own meat brands ・Local farmers and town butchers can develop their own cell-based meat recipe. ・Brand ownership opportunity opens up for local farmers. ・Hobbyists come up with unique meat recipe.
Celebrity Star Cow branded beef The cell source Star Cow is alive Scan the QR code printed on package Watch the Star Cow
Less cows Bioreactors in the backyard Many cows Cows are the product Cells or meat is the product Traditional farmscape Cell-ag farmscape The Cell-Ag Farmscape Less number of cows mean less cleaning, feeding, waste treatment More facility management and cell culture protocol dev., as in brewery
Business models of meat in cell-ag era Fabless farmers Contract farmer Lend cows Meat brewery Sends cells Sells meat Fabless farmer Farmers Cell licensing Cell bank Sends cells Food companies Sells meat Shared recipe Recipe website Warehouse People cell culture protocols Domestic culture vat Down- load Farmers Stores cells Sends cells Cell-Ag Firms People Cell-ag firm “New farmer” People Sells meat Food companies Meat brewery Sells cells More info↓ https://www.slideshare. net/2co/the-cellular-agri culture-farmscape
Myoblast grown at 200t scale Looks like a beer factory? Future: Meat brewery
Steak brewery Single use culture bags⇒ Steak grows inside⇒ Ready to ship⇒
Seafood by cellular agriculture Cell culture at room temperature Small ponds as cell source
Culture media produced by microalgae Short food miles Farm high-rise Cell-ag towers
Farmscape, A.D.2203 Vertical farms Vast land reverted to nature Artificial photosynthesis Cellular agriculture manufacturing industry complex
Martian food production facility
Orbital Zero-G Farm
Creative support for artists “I deal with Ideal Meal”　by Yamada Theta https://twitter.com/yamada_theta https://www.pixiv.net/member_illust.php?mode=medium&illust_id=66586574 　 https://note.mu/yamada_theta/n/n243f40556409 《Schroedinger’s Tiger》 by Michi Okada http://tobira-project.info/blog/2017_okada.html http://michiokada.tumblr.com/
Miyo-san ”Myosin” age.20・164cm Chemical engineering student intern at Mars Huygens Crater cellular agriculture facility. Aco-chan ”Actin” age.13・149cm Helps elder sister Miyo in her extracurricular activity at Mars Colonists Middle School. Miyo & Aco
Miyo-san (Chibi)
Aco-chan (Chibi)
Visual novel source files Source files https://drive.google.com/drive/u/0/folders/0B0ShPzNziL05THlwU2IwTTNueHM (Feel free to produce alternative scenarios, derivative works, add characters, etc. under CC-BY-NC license.) “Miyo-san” and “Aco-chan” whole body and face PNG images and sci-fi themed background images available for manga and VN’s. MiyoAco
Perhaps, someday in the future on Mars….
Zine's (DIY cell culture manuals) Distributed at Comic Market, COMITIA, TechBookFest, etc. Also available from MelonBooks Sold out 2019 winter DIY cell culture manual https://www.melonbooks.co.j p/detail/detail.php?product_i d=616192 https://www.slideshare.net/YukiH anyu/ss-85789390 https://www.slideshare.net/YukiH anyu/ss-79853068 https://www.slideshare.net/YukiH anyu/diy-85789299 Online materials (slideshare)
Members participate in their respective expertise (experiments, gatherings, art projects etc.) - no defined “membership” #Food Security #Food Miles #Regulations #Cooking #Culture & thoughts #History #Food safety #Life ethics #Animal welfare #Regenerative medicine #Tissue engineering #Bioreactor #Culture medium #Commercialization #LCA #R&D #Soc.&Econ. #Global collab. #Space #Art Shojinmeat Project “Distributed Clusters” All welcome: Please join from Shojinmeat Project English Slack Channel https://medium.com/@jaysonvdw/announcement-english-slack-channel-for-the-shojinmeat-project-a8 00e3886799
Some past questions (part 1) Q:　I would like to participate A:　There is no formal definition of membership. Please Tweet to us or join slack channel. Q:　I’m not a biologist but would like to join A:　All disciplines welcome - multiple journalism, art & other non-sci/tech projects are ongoing Q:　Where are carbon and nitrogen sourced? A:　Culture medium contains sugar and amino acids. Amino acid is sourced from yeast which feed on sugar. In future, artificial photosynthesis and nitrogen fixation may take this role. Q:　Is fungal farming cellular agriculture? A:　Depends on if cell culture procedure is involved in the process. Cultivation of entire fungal body would be conventional farming. Q:　Is cell-based meat GMO? A:　Gene editigin is not required for cell culture. In future, “designer meat” such as “allergen-free meat” may require gene editing. Q:　Is it tasty? A:　It’s pasty (for now). In future, advanced tissue engineering may enable complete reconstruction of meat taste and texture and even go beyond. Q:　How quickly would “meat” grow? A:　Cells multiply every 1½~2 days. 1E5 cells grow to visible size in 20~30 days. Flow process in factories may enable continuous production. Q:　What will happen to farm animals? A:　They don’t disappear as starter cells would still be required, but their number may decline.
Some past questions (part 2) Q: Could meat be cocultured with probiotics? A: Unknown. It depends on their growth rates and mutual effect of cell metabolites. Q: Why would people eat cell-based meat?　 A: Initially, vegans and environmentally-conscious would consume, but at the end, taste, price and convenience would decide. Q: My kids should try DIY cell culture A: Hardware and methods are open for public. Please contact us for details. Q: Desktop clean bench may help? A: Egg white can suppress mould growth, but a clean bench always help. Q: What will happen to farmers? A: Farmers may gain an opportunity to build a brand around his/her cow cell and meat culture recipe, but it depends on what business model cellular agriculture companies take in the future. Q: Is cell-based meat Halal/Kosher? A: It may potentially become Halal/Kosher under certain conditions, according to a publication: https://www.ncbi.nlm.nih.gov/pubmed/28456853 Q: Is it vegan? A: Depends on technological maturity. Our DIY procedure uses egg white as antimycotic and therefore not vegan. In future, it depends on the exact procedure of cell acquisition.
References for common facts More than 80% of arable land, or 20% of Earth’s land surface is used for meat production. It is also responsible for 18% of total greenhouse gas emissions. “Reducing food’s environmental impacts through producers and consumers” Poore, Nemecek, Science 2018 http://science.sciencemag.org/content/360/6392/987 More than 80% of arable land is used for meat in some ways. FAO www.fao.org/animal-production/en/ 20~30% of all freshwater is used for farm animals “The water footprint of humanity”, Hoekstra, Mekonnen, PNAS 2012, http://www.pnas.org/content/109/9/3232 Energy and protein feed-to-food conversion efficiency is 3% and 4% respectively. “Energy and protein feed-to-food conversion efficiencies in the US and potential food security gains from dietary changes”, Shepon, Eschel et al, IOP Science 2016 http://iopscience.iop.org/article/10.1088/1748-9326/11/10/105002/meta 1kg of beef requires 25kg of feed and 15000L of freshwater Hoekstra, Mekonnen, UNESCO IHE 2010 “The green, blue and grey water footprint of farm animals and animal products” http://waterfootprint.org/media/downloads/Report-48-WaterFootprint-AnimalProducts-V ol1.pdf 90％ of soybean and 40% of crops are consumed as feed How to feed the world 2050 High-level expert forum, Rome 12-13 Oct.2009 http://www.fao.org/fileadmin/templates/wsfs/docs/Issues_papers/HLEF2050_Global_Ag riculture.pdf World’s feed can support 3.5B people if consumed as food. 1kg of beef requires 12~24kg of feed. “Redefining agricultural yields: from tonnes to people nourished per hectare”, Cassidy, West e tal, IOP Science, Environmental Research Letters, 2013, http://iopscience.iop.org/article/10.1088/1748-9326/8/3/034015/meta 1kg of poultry requires 2~4kg of feed Meat or wheat for the next millennium? Alternative futures for world cereal and meat consumption https://pdfs.semanticscholar.org/ae0c/a5b5556e57a2c4cf904b218d0edbac2c32d9.pdf 80% of antibiotics are used in farm animals. “The antibiotic resistance crisis: part 1: causes and threats.”, Ventola, NCBI 2015, https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4378521/ Organic farming requires larger land area per unit yield. “Is organic really better for the environment than conventional agriculture?”, Ritchie, OWID 2017, https://ourworldindata.org/is-organic-agriculture-better-for-the-environment
Contacts for businesses “Shojinmeat Project” is a citizen science community. Please contact IntegriCulture Inc. for industry & business collaboration. Please contact CAIC for market information & industry consultation. And welcome to the Shojinmeat Project to passionate individuals! http://integriculture.jp http://cellagri.org
Get involved in cellular agriculture! DIY Bio, Make. , public communication Tuesdays 20:00 FabCafeTokyo 2F https://fabcafe.com/tokyo/access/ @shojinmeat Join Shojinmeat English Slack channel https://medium.com/@jaysonvdw/anno uncement-english-slack-channel-for-th e-shojinmeat-project-a800e3886799
Acknowledgements All Shojinmeat Project participants! Patrons at Campfire Crowdfunding

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