Progress 01/01/24 to 12/31/24
Outputs
Target Audience:-Academic communities across disciplines: e.g. the integrative Center for Cultured Meat and Alternative Proteins, the University of California at Davis; theUniversity of Minnesota, Plant Protein Innovation Center (PPIC); Johns Hopkins University School of Medicine, Department of Cell Biology; Laval University,Institute of Nutrition and Functional Foods, Pôle bioalimentaire, Laval, Canada;Penn State University, "Boosting Bioethics & Bioprinting" Webinar Series hosted by the Huck Institute of the Life Sciences, the College of Engineering, the Rock Ethics Institute, & the Penn State Law,University Park, USA; Carnegie Mellon University, Department of Bioengineering Seminar, Pittsburgh, USA -Legislators:California State Capitol, the Alt Protein Working Group of Assemblymembers and Congress members, Sacramento, CA. -Students: Graduate student trainees are becoming experts in the field of cellular agriculture, including trainees in the Rowat lab, Stephanie Kawecki (Postdoctoral Research Fellow, Integrative Biology & Physiology); Corinne Smith and Rebecca Cohen (PhD candidates, Bioengineering); Qingwen Xie (Masters student graduated 06/2024; and subsequently Research Associate, Integrative Biology & Physiology); Emily Cheng (Research Assistant and Undergraduate Student in Molecular, Cellular, and Developmental Biology, Food Studies graduated 12/2024; and subsequently Research Associate). Undergraduate students are receiving hands-on research training experiences including Ester Fridman (Sociology), Adam Nguyen (Anthropology, Food Studies) and Zoe Yee (Microbiology, Immunology, and Molecular Genetics and Sustainable Los Angeles Grand Challenge (SLAGC) Undergraduate Research Scholar).The Future Food Fellows training program additionally provides opportunities for graduate students, undergraduate student researchers, and postdoctoral fellows who lead the program as mentors.The program isdesigned to provide tailored research experiences and industry networking for graduate students who are passionate about the future of food. -Industry: PI Rowat has explored collaborative opportunities with companies including CP Kelco and Umami Bioworks.Our technology is being considered for licensing to companies in the space. In addition, PI Rowat interacts regularly with companies, e.g., through theSouthern California Food Industry Conference,the integrative Center for Cultured Meat and Alternative Proteins, the University of California at Davis, theUniversity of Minnesota, and the Plant Protein Innovation Center (PPIC). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has provided the following training and professional development opportunities: •One-on-one research mentorship of PI Rowat with trainees Corinne Smith, Stephanie Kawecki, Kathleen Chen, Rebecca Cohen, Qingwen Xie, and Emily Cheng. • Dr. Kawecki and Corinne Smith presented posters at theInternational Scientific Conference on Cultured Meat (ISCCM) 2024, Maastricht, the Netherlands. •The project provided support for previous Bioengineering graduate student researcher, Stephanie Kawecki, as a postdoctoral researcher. • Support for undergraduate research opportunities:Ester Fridman (Sociology), Adam Nguyen (Anthropology, Food Studies); Emily Cheng (Molecular, Cellular, and Developmental Biology, Food Studies); Zoe Yee (Microbiology, Immunology, and Molecular Genetics and Sustainable Los Angeles Grand Challenge (SLAGC) Undergraduate Research Scholar); and Sydney Adler (Chemical Engineering). •Mentorship provided by PI Rowat for graduate and undergraduate students to lead an Alt Protein Project initiative at UCLA. •The project attracted additional funding from the State of California, which has enabled additional training experiences. Funding from the State of California to UCLA, UC Davis, and UC Berkeley for research in alternative proteins and cultured meat (total of $5M, with $1.6M awarded to UCLA, 2023-2025).The funding was allotted to UCLA after PI Rowat's research advances in cultured meat attracted the attention of state assemblymembers.This funding has supported the following: -To strengthen research and education around cultured meat at UCLA, PI Rowat hired Research Associate Dr. Muriel Vernon, who engaged with industry to conduct an industry-needs survey. In the current reporting period, we wrote a manuscript, which is currently under review for peer-reviewed publication; this will provide a model for institutions to design multidisciplinary training opportunities for students (Vernon et al, Manuscript under review). - In Fall 2024, PI Rowat launched the UCLA Science&Food Future Food Fellows graduate training fellowship program. The program aims to build community among a cohort of graduate student researchers who are dedicated to sustainable food futures; the program will strengthen academia-industry relationships while addressing workforce demands from the industry for trained individuals. The program structure was guided by findings from studies led by Dr. Muriel Vernon, which revealed successful elements of the New Harvest fellow program and also industry needs. The ultimate goal of the Future Food Fellows program is to advance research in cultivated meat andto address workforce demands from the industry for trained individuals. -State funds have furthermore supported the development of critical infrastructure, including a nanoindentation instrument to quantify the mechanical properties of cells and tissues, and a larger-scale bioreactor, which will enable us to test proof-of-concept findings in a larger stirred-tank bioreactor context. •To further establish the global cellular agriculture research community, PI Rowat contributed to the development of the firstGordon Research Conference in the field of Future Foods together with Co-Chairs David Kaplan, Mette Lübeck, Girish Ganyal. Our proposal to the GRCConference Evaluation Committee (CEC)received positive reviews and the first conference has been approved for January 2026, 'Foods of the Future: Science and Engineering Approaches'. •PI Rowat is also gaining training and professional development opportunities through providing educational opportunities and communicating Rowat lab research updates to state legislators and assemblymembers. How have the results been disseminated to communities of interest?PEER-REVIEWED PUBLICATIONS: Kawecki NS,Chen KK,Smith CS, Xie Q, Cohen JM,Rowat AC(2024)Scalable Processes for Culturing Meat Using Edible Scaffolds.Annu Rev Food Sci Tech. 15: 241-264. INVITED LECTURES BY PI ROWAT (2024): PI Rowat received the following speaker invitations: Carnegie Mellon University, Department of Bioengineering Seminar, Pittsburgh, USA California State Capitol, Research Updates for the Alt Protein Working Group of Assemblymembers and Congress members, Sacramento, CA Southern California Food Industry Conference (SCIFIC) 2024 - The Future of Food Science [virtual] University of Minnesota, Plant Protein Innovation Center (PPIC) 5thAnnual Research Spotlight Meeting, Minneapolis, USA Calico, Mechanosensation Mini-Symposium, San Francisco, USA University of California, Davis, integrative Center for Alternatives Meat and Proteins (iCAMP) Partners Retreat, Davis, USA Johns Hopkins University School of Medicine, Cell Biology Colloquium, Baltimore, USA Penn State University, "Boosting Bioethics & Bioprinting" Webinar Series hosted by the Huck Institute of the Life Sciences, the College of Engineering, the Rock Ethics Institute, & the Penn State Law,University Park, USA [webinar] Laval University,Institute of Nutrition and Functional Foods, Pôle bioalimentaire, Laval, Canada [webinar] FEATURED IN THE MEDIA: Lab research featured by the Big 10 Production Team (aired at UCLA basketball games 11/20/2024 and 11/30/2024) Lab research featured in Spectrum News 1 - Innovations for a Sustainable Climate:https://spectrumnews1.com/ca/southern-california/news/2024/02/20/innovations-for-a-sustainable-climate--a-spectrum-news-1-special?cid=id-app15_m-share_s-web_cmp-app_launch_august2020_c-producer_posts_po-organic(Feb 2024) Featured in the Washington Post, "What a lab-made meat-rice hybrid says about the future of food" (KK Cho, February 2024) Featured in the UCLA Newsroom, 'UCLA's inaugural Future of Food Fellows poised to advance the field of cellular agriculture' (L Berbeo, November 2024) Featured in UCLA College Magazine, 'Lab to Life: How UCLA College experts are translating the power of science into real-world health solutions' (L Berbeo, December 2024) Featured in UCLA Newsroom, 'Amy Rowat named Allen Distinguished Investigator' (H Ober, December 2024) Featured by the Allen Institute, 'The Paul G. Allen Family Foundation Awards $9 Million to new Allen Distinguished Investigators' (A Addrisi, December 2024) What do you plan to do during the next reporting period to accomplish the goals?In the next reporting period, we will test our edible microcarriers to support the growth of bovine muscle and fat tissues, and generation of cultivated beef with tunable fat content.We will characterize the textural and nutritional properties of the cultivated beef and fat.We requested a no-cost extension for the award because the planned isolation of bovine cells from beef tissues has delayed our experimental progress.We recently received primary bovine, ovine and porcine satellite muscle cells and pre-adipocytes from the company, OpoBio.
Impacts
What was accomplished under these goals?
In the annual reporting period, we have made strong progress towards our goal of producing a cultured meat burger with defined nutrition and sensory properties to meet consumer needs.In the last year, we have: 1) Extended our approach to culturing adipose tissue using edible microcarrier scaffolds to porcine pre-adipocytes and further discovered that the physical properties of microcarrier scaffolds can be tuned to accelerate lipid accumulation in adipose microtissues (Objective 2, Kawecki, Cohen et al,Manuscript in preparation). 2) Tested the nutritional and textural properties of cultured beef with varying amounts of cultivated fat. Our current work builds on our findings from the previous reporting period that edible microcarriers can seed the growth of microtissue aggregates, which provide building blocks for assembling larger-scale tissue constructs (Objective 3, Kawecki et al,Food Research International). Objective 1: Develop edible microcarrier scaffolds to support cultured muscle microtissue. Major activities completed:In a previous reporting period, we reported on fabricating microcarrier scaffolds to support the growth of muscle microtissue (Norris et al,Biomaterials, 2022). Data collected:See Norris et al,Biomaterials,2022. Summary and discussion of results:See Norris et al,Biomaterials,2022. Key outcomes:1)Edible microcarriers can support the generation of bovine and murine myogenic microtissues in suspension culture.2)Results show the potential of using edible microcarriers with tunable stiffness and topology to reduce the volume fraction of scaffolds in the final cultured meat product. Objective 2: Develop edible microcarrier scaffolds to support cultured fat microtissue with desired lipid content. Major activities completed:We designed microbead scaffolds withE~4 kPa, which mimics the stiffness of adipose tissue (Kawecki et al,Food Res Int, 2023). We can also size-filter microbeads to generate populations of microbeads with diameter ranging from < 20 µm to 150 µm (as in Norris et al,Biomaterials, 2022).We proliferate and differentiate pre-adipocytes, and then measure lipid accumulation using confocal microscopy and quantitative image analysis and expression of adipogenic markers (PPARg, CEBPα) using RT-PCR (as in Kawecki et al,Food Res Int, 2023). In the current reporting period, we extended this approach to demonstrate the ability to culture porcine fat. Data collected:We report lipid levels over up to 14 days, observing increased accumulation of intracellular lipids, as evidenced by the increased signal of LipidTox, which is a marker for neutral lipids. We further quantify adipogenesis at the transcript level using qRT-PCR.To quantify levels of adipogenic markers in cultured porcine adipose tissue, we additionally conducted RNAseq analysis. Summary and discussion of results:Findings reveal that we can accelerate the growth and maturation of adipose tissue in vitro by tuning the size of edible microbeads. We find that edible microbeads support cell proliferation and adipogenesis as quantified by increased lipid area across murine 3T3-L1 adipocytes, rabbit RbPreAd cells, and porcine pre-adipocytes. Analysis of theprofiles of lipid speciesin cultured meat samples revealed that triglycerides account for the largest proportion oflipid species(53-55%), with the ratio of triglycerides to other lipids being similar to that found in conventional meat. These findings highlight the potential of cultured meat to replicate the lipid profile ofconventional cuts of meat. Key outcomes: 1)Edible microbeads can support the growth of fat microtissues derived from different cell types, including primary rabbit cells, porcine pre-adipocytes, as well as murine cell lines in suspension culture.2)Tuning microbead physical properties can increase the rate of lipid accumulation during adipogenesis. Specifically, we discovered that the size of edible microcarriers can be tuned to modulate adipogenesis in murine, rabbit, and porcine fat tissues.3)Analysis of the profiles of lipid species in cultured meat shows the potential of cultured meat to replicate the lipid profile of conventional cuts of meat. Objective 3: Produce cultured ground beef with desired qualities using edible microcarrier scaffolds. Major activities completed:In the previous reporting period, we discovered that myogenic and adipogenic microtissues adhere to each other on timescales of ~6-10 hours, which enables the production of multicomponent tissues (Kawecki et alFood Res Int2023).In the current reporting period, we have been developing protocols to quantify macroscopic properties of cultured ground beef using edible microcarrier scaffolds using techniques including rheology, differential scanning calorimetry (DSC), lipidomics, and proteomics. Data collected:We have developed protocols to quantify the nutritional and cooking properties of marbled cultured meat by measuring macronutrient content (protein, total lipids, carbohydrates), cooking loss, and rheological properties as a function of temperature.Pilot data include shear and loss moduli as a function of temperature (rheology), lipidomics, and heat capacity as a function of temperature (DSC). Summary and discussion of results:Pilot data reveal that cultivated muscle/fat tissue cultured on edible microcarrier scaffolds shows properties of a viscoelastic material that has predominantly solid-like behavior at temperatures < 40C.These findings build on our discovery from the previous reporting period that edible microcarrier scaffolds can seed the growth of muscle and fat microtissues, which provide the building blocks for assembling larger, multicomponent tissues.Our preliminary nutritional analysis reveals similar protein content as Wagyu beef steaks.We are currently undertaking more detailed analysis of the adipose microtissues, with the goal to build ground beef with tunable fat content. Key outcomes: 1)Engineered edible scaffolds provide the foundation for structuring and organizing both myogenic and adipogenic cells and microtissues; the resultant multicomponent tissue constructs form by spontaneous adhesion of microtissues on ~6-10 hour timescales, which is mediated by cells and does not require the use of additional crosslinkers; this can reduce the number of post-processing steps and required reagents.2)Multicomponent tissue composed of different types of cells and scaffolds can be generated through the spontaneous adhesion of microtissues.3)The resultant multicomponent tissues show behavior of a viscoelastic solid material, similar to conventional meat (skeletal muscle).4)The resultant multi-component, myogenic-adipogenic tissues (cultured meat) have a similar protein content as Wagyu beef steak.5)Analysis of the profiles of lipid species in cultured meat shows the potential of cultured meat to replicate the lipid profile of conventional cuts of meat.6)Edible microcarriers support the production of myogenic and adipogenic microtissues that are derived from different species (bovine, porcine, rabbit, murine).7)Upon cooking, the resultant multicomponent tissues (cultured meat) show slightly more cooking loss but also exhibit signs of browning characteristic of Maillard reactions.8)The scalable approach that we describe to generate edible microcarriers and the resultant muscle and adipose microtissues have potential to contribute to efficient, cost-effective cultured meat production, which could provide a complementary alternative for protein production that ultimately could help to increase the resilience of future food systems.
Publications
- Type:
Peer Reviewed Journal Articles
Status:
Published
Year Published:
2024
Citation:
Kawecki NS, Chen KK, Smith CS, Xie Q, Cohen JM, Rowat AC (2024) Scalable Processes for Culturing Meat Using Edible Scaffolds. Annu Rev Food Sci Tech. 15: 241-264.
|
Progress 01/01/23 to 12/31/23
Outputs
Target Audience:-Academic communities across disciplines: e.g. the Human Frontiers Science Program meeting; the integrative Center for Cultured Meat and Alternative Proteins, the University of California at Davis. -Students: Graduate student trainees are becoming experts in the field of cellular agriculture, including trainees in the Rowat lab, Stephanie Kawecki (PhD candidate, Bioengineering); Corinne Smith (PhD candidate, Bioengineering); Kathleen Chen (PhD candidate, Chemistry); Qingwen Xie (Masters student, Physiology). In the current reporting period, both Dr. Stephanie Kawecki and Dr. Kathleen Chen graduated from their respective graduate programs. Dr. Kawecki is currently a postdoctoral researcher in the Rowat Lab. Undergraduate students are receiving hands-on research training experiences including Ester Fridman (Sociology), Adam Nguyen (Anthropology, Food Studies); Emily Cheng (Molecular, Cellular, and Developmental Biology, Food Studies); and Zoe Yee (Microbiology, Immunology, and Molecular Genetics and Sustainable Los Angeles Grand Challenge (SLAGC) Undergraduate Research Scholar). We are currently working to launch a pilot graduate student fellowship training program designed to provide tailored research experiences and industry networking for graduate students who are passionate about the future of food. -Companies: PI Rowat has initiated collaborative opportunities with companies including CP Kelco. Our technology is being considered for licensing to companies in the space. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has provided the following training and professional development opportunities: •One-on-one research mentorship of PI Rowat with trainees Corinne Smith, Stephanie Kawecki, and Qingwen Xie. • Dr. Kawecki was invited to give a talk at the Society for Biomaterials meeting (2023) and the International Scientific Conference on Cultured Meat (ISCCM) 2023, Maastricht, the Netherlands. •The project provided support for previous Bioengineering graduate student researcher, Stephanie Kawecki, who recently defended her thesis in December 2023. The project continues to provide training for Dr. Stephanie Kawecki, who is now a postdoctoral researcher in the group. • Support for undergraduate research opportunities: Ester Fridman (Sociology), Adam Nguyen (Anthropology, Food Studies); Emily Cheng (Molecular, Cellular, and Developmental Biology, Food Studies); and Zoe Yee (Microbiology, Immunology, and Molecular Genetics and Sustainable Los Angeles Grand Challenge (SLAGC) Undergraduate Research Scholar). Ester Fridman (Sociology), Adam Nguyen (Anthropology, Food Studies); Emily Cheng (Molecular, Cellular, and Developmental Biology, Food Studies); and Zoe Yee (Microbiology, Immunology, and Molecular Genetics and Sustainable Los Angeles Grand Challenge (SLAGC) Undergraduate Research Scholar). •Mentorship provided by PI Rowat for graduate and undergraduate students to lead an Alt Protein Project initiative at UCLA. •The project has further attracted additional funding from the State of California, which is enabling additional training experiences. Funding from the State of California to UCLA, UC Davis, and UC Berkeley for research in alternative proteins and cultured meat (total of $5M, with $1.6M awarded to UCLA, 2023-2025). The funding was allotted to UCLA after PI Rowat's research advances in cultured meat attracted the attention of state assemblymembers. To strengthen research and education around cultured meat at UCLA, PI Rowat hired Research Associate Dr. Muriel Vernon, who engaged with industry to conduct an industry-needs survey; findings from the survey are being used to design the graduate fellows program. The ultimate goal of the graduate fellows program is to advance research in cultivated meat andto address workforce demands from the industry for trained individuals.PI Rowat is also gaining training and professional development opportunities through providing educational opportunities and communicating Rowat lab research updates to state legislators and assemblymembers. How have the results been disseminated to communities of interest?PEER-REVIEWED PUBLICATIONS(# corresponding author; bold denotes Rowat Lab members): Kawecki NS, Chen KK, Smith CS, Xie Q, Cohen JM, Rowat AC#. Scalable Processes for Culturing Meat Using Edible Scaffolds. Annu Rev Food Sci Tech. In press Kawecki NS, Norris SCP, Xu Y, Wu Y, Davis AR, Fridman E, Chen KK, Crosbie RH, Garmyn A, Li S, Mason T, Rowat AC# (2023) Engineering multicomponent tissue by spontaneous adhesion ofmyogenic and adipogenic microtissues cultured with customized scaffolds. Food Research International. 172: 113080 Kawecki NS, Rowat AC# (2023) Future Pathways in Biosynthesis: Livestock Tissue Culture in 'The Protein Transition'. Eds. Pyett S, van Mierlo B, Trindade L, de Vet E, Welch D, van Zanten H, de Jong L, and van den Top M [Invited Book Chapter] INVITED LECTURES BY PI ROWAT (2023): PI Rowat received the following speaker invitations: Integrative Center for Alternative Meat and Protein (iCAMP), University of California, Davis, USA (Keynote Speaker) Center for Arts & Humanities in Medicine, Cedars Sinai, Los Angeles, USA [virtual] International Scientific Conference on Cultured Meat (ISCCM) 2023, Maastricht, the Netherlands (Keynote Speaker) Princeton University, Princeton Food Project, Princeton, USA Human Frontiers Science Program Organization, High-Level Summit and International Scientific Symposium on Fundamental Life Science Meets Climate, Environment and Sustainability, Paris, France (Invited talk and Invited panelist, '21st Century the 'Age of Biology' Challenges and Opportunities for Sustainability with Marcia McNutt and Mona Nemer) FEATURED IN THE MEDIA: Featured in The Chronicle of Philanthropy, "Does alternative meat need philanthropy to take it beyond?' (E. Stiffman, October 2023) Featured in the Atlantic, "Open your mind tounicornmeat" (A. Lowrie, July 2023) Featured in Nature, "Lab-grown meat: the science of turning cells into steaks and nuggets" (N. Jones, July 2023) UNDERGRADUATE AND HIGH SCHOOL EDUCATION: PI Rowat featured cultured meat during one week of the interdisciplinary undergraduate class that she developed and taught in spring 2023, FOOD STUDIES 181, Special Topics: Perspectives on Food and Society. Students engaged in critical discussion around livestock agriculture, from methods in regenerative agriculture to cellular agriculture. PI Rowat initiated and contributed to the development of the undergraduate class, 'Cellular Agriculture' (piloting Spring 2024). PI Rowat developed an interactive presentation, which she presented to the 2nd and 3rd grade students of the Santa Monica Alternative School House, Santa Monica, CA, USA. PI Rowat was an invited lecturer for the San Diego State University, Seminar for Nanotechnology Students, San Diego, USA [virtual]. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
There is urgent need to diversify methods for protein production so that our future food system will be resilient to disruptions due to the impacts of climate variability, natural disasters, and future epidemics. One potential emerging approach uses tissue culture methods to enable cultured meat. The goal of this specific project is to produce a cultured meat burger with defined fat content to meet different consumer needs, and to identify supplements that enable us to tune the lipid composition of cultured meat towards achieving nutritious and flavorful cultured meat. In the annual reporting period, we have made strong progress towards our goal of producing a cultured meat burger with defined nutrition and sensory properties to meet consumer needs. Our strategy to achieving this goal is to develop edible 'microcarrier' scaffolds that support the growth of cultured tissues in a large vessel or bioreactor, which has potential to scale up tissue culture that is required for food production. In the last year, we have: 1) Developed edible microcarrier scaffolds to support cultured fat microtissue (Objective 2; Kawecki et al, Food Research International) and further discovered that the physical properties of microcarrier scaffolds can be tuned to accelerate lipid accumulation in adipose microtissues (Objective 2, Kawecki et al, Manuscript in progress); and 2) Demonstrated that the microtissues seeded on edible scaffolds provide building blocks for assembling larger-scale tissue constructs (Objective 3, Kawecki et al, Food Research International). We continue to make progress towards translating this technology, and are currently in discussions about licensing and sponsored research agreements with companies in the space. Objective 1: Develop edible microcarrier scaffolds to support cultured muscle microtissue. Major activities completed: In the previous reporting period, we reported on fabricating microcarrier scaffolds to support the growth of muscle microtissue (Norris et al, Biomaterials, 2022). Data collected: See Norris et al, Biomaterials, 2022. Summary and discussion of results: See Norris et al, Biomaterials, 2022. Key outcomes: 1) Edible microcarriers can support the generation of bovine and murine myogenic microtissues in suspension culture. 2) Results show the potential of using edible microcarriers with tunable stiffness and topology to reduce the volume fraction of scaffolds in the final cultured meat product. Objective 2: Develop edible microcarrier scaffolds to support cultured fat microtissue with desired lipid content. Major activities completed: We designed microbead scaffolds with E ~4 kPa, which mimics the stiffness of adipose tissue (Kawecki et al, Food Res Int, 2023). We can also size-filter microbeads to generate populations of microbeads with diameter ranging from < 20 µm to 150 µm (as in Norris et al, Biomaterials, 2022). We proliferate and differentiate pre-adipocytes, and then measure lipid accumulation using confocal microscopy and quantitative image analysis and expression of adipogenic markers (PPARg, CEBPα) using RT-PCR (as in Kawecki et al, Food Res Int, 2023). Data collected: We report lipid levels over up to 14 days, observing increased accumulation of intracellular lipids, as evidenced by the increased signal of LipidTox, which is a marker for neutral lipids. We further quantify adipogenesis at the transcript level using qRT-PCR. Summary and discussion of results: Findings reveal that we can accelerate the growth and maturation of adipose tissue in vitro by tuning the size of edible microbeads. We find that edible microbeads support cell proliferation and adipogenesis as quantified by increased lipid area across murine 3T3-L1 adipocytes, rabbit RbPreAd cells, and porcine pre-adipocytes. Key outcomes: 1) Edible microbeads can support the growth of fat microtissues derived from different cell types, including primary rabbit cells, porcine pre-adipocytes, as well as murine cell lines in suspension culture. 2) Tuning microbead physical properties can increase the rate of lipid accumulation during adipogenesis. Objective 3: Produce cultured ground beef with desired qualities using edible microcarrier scaffolds. Major activities completed: To investigate the spontaneous adhesion of myogenic and adipogenic microtissues across cell and scaffold types, we developed an adhesion assay. We also investigate the adhesion of myogenic and adipogenic microtissues cultured with plant-based scaffolds. To assess the timescale of adhesion, we measure the adhesion index at 0, 6, 12, and 18 hour timepoints. We characterize the mechanical properties of the multicomponent tissue--which contains both myogenic and adipogenic microtissues--using a parallel plate rheometer and tensile analysis. We also quantify protein content and assessed cooking loss. Data collected: To quantify microtissue adhesion, we mechanically perturb a layered stack of myogenic and adipogenic microtissues by rigorous nutation and then assess how intact the microtissue stack remains after nutating by quantifying the projected surface area of microtissue that is displaced after the mechanical perturbation. To begin to quantify the nutritional properties of the marbled cultured meat, we measure total protein content. To assess cookability, we cook the cultured meat samples in a clamshell grill and quantify the loss of aqueous and lipid components in a sample during the cooking process. Summary and discussion of results: The modular approach that we developed to generate cultured meat that contains both muscle and fat components--or myogenic and adipogenic microtissues--enables separate growth of myogenic and adipogenic microtissues, which can adhere to each other within 6-10 hours to form cohesive, multicomponent tissue constructs. The multicomponent tissues show qualitatively similar mechanical behaviors to conventional Wagyu beef steak. Further tuning the post-harvesting steps can be explored to achieve more similar textures as conventional meats. Both rabbit and mouse marbled cultured meat samples show a median cooking loss of ~30%, which is higher although not statistically different than the ~20% cooking loss observed for the Wagyu steak samples. We found no statistically significant differences in protein content for both cultured rabbit and murine meat samples compared to conventional Wagyu steaks. Key outcomes: 1) Engineered edible scaffolds with customized physical properties provide the foundation for structuring and organizing both myogenic and adipogenic cells and microtissues; the resultant multicomponent tissue constructs form by spontaneous adhesion of microtissues on ~6-10 hour timescales, which is mediated by cells and does not require the use of additional crosslinkers; this can reduce the number of post-processing steps and required reagents. 2) Multicomponent tissue composed of different types of cells and scaffolds can be generated through the spontaneous adhesion of microtissues. 3) The resultant multicomponent, myogenic-adipogenic tissues (cultured meat) have similar protein content as Wagyu beef steak. 4) Edible microcarriers support the production of myogenic and adipogenic microtissues that are derived from different species (bovine, porcine, rabbit, murine). 5) Upon cooking, the resultant multicomponent tissues (cultured meat) show slightly more cooking loss but also exhibit signs of browning characteristic of Maillard reactions. 6) The scalable approach that we describe to generate edible microcarriers and the resultant muscle and adipose microtissues have potential to contribute to efficient, cost-effective cultured meat production, which could provide a complementary alternative for protein production that ultimately could help to increase the resilience of future food systems.
Publications
- Type:
Book Chapters
Status:
Published
Year Published:
2023
Citation:
Kawecki NS, Rowat AC# (2023) Future Pathways in Biosynthesis: Livestock Tissue Culture in The Protein Transition. Eds. Pyett S, van Mierlo B, Trindade L, de Vet E, Welch D, van Zanten H, de Jong L, and van den Top M [Invited Book Chapter]
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Kawecki NS, Norris SCP, Xu Y, Wu Y, Davis AR, Fridman E, Chen KK, Crosbie RH, Garmyn A, Li S, Mason T, Rowat AC# (2023) Engineering multicomponent tissue by spontaneous adhesion of myogenic and adipogenic microtissues cultured with customized scaffolds. Food Research International. 172: 113080
- Type:
Journal Articles
Status:
Accepted
Year Published:
2023
Citation:
Kawecki NS, Chen KK, Smith CS, Xie Q, Cohen JM, Rowat AC#. Scalable Processes for Culturing Meat Using Edible Scaffolds. Annu Rev Food Sci Tech. In press
|
Progress 01/01/22 to 12/31/22
Outputs
Target Audience:-Academic communities across disciplines: e.g. the Biophysical Society meeting; the Department of Food Science & Nutrition, Michigan State University -Students: Graduate student trainees are becoming experts in the field of cellular agriculture, including trainees in the Rowat lab, Stephanie Kawecki (PhD candidate, Bioengineering); Corinne Smith (PhD candidate, Bioengineering); Kathleen Chen (PhD candidate, Chemistry); Qingwen Xie (Masters student, Physiology). Undergraduate students are receiving hands-on research training experiences including Ester Fridman (Sociology). We are currently working to design practicum experiences for students so they could gain perspectives from industry. -Companies: PI Rowat has initiated collaborative opportunities with companies including CP Kelco and Geltor.Our technology is being considered for licensing to a startup. Efforts -Formal classroom instruction: PI Rowat developed and instructed the class Food Studies 181,Special Topics: Perspectives on Food and Society,which included asection on the future of food.In addition, PI Rowat has delivered lectures on cultured meat to students at the 50thannual Geilo Winter School, Geilo, Norway; students in the classBio290 "The Cellular Agriculture Revolution", at theUniversity of North Carolina, Chapel Hill. -Development of curriculum: Our team has developed innovative approaches to engaging students in the future of food using hands-on demonstrations and tastings in the teaching kitchen at UCLA.As part of the Food Studies 181 class taught by PI Rowat in Spring 2022, we developed an interactive tasting where Chef Julia Rhoton prepared plant-based burgers, beef burgers, and bean burgers.Students tasted and discussed factors that impacted their choice and preference of burger, which built on in-class discussions and lectures by PI Rowat and collaborator and advisor at Michigan State University, Dr. Jason Rowntree. -Outreach: PI Rowat was featured in the media by: •California NanoSystems Institute, "Bruin biophysicist's research pushes forward development of cultured meat" (W. Lewis, Aug 2022) •Featured in UCLA Newsroom, "UCLA scientists bring cultured meat closer to your kitchen table" (H. Ober, Aug 2022) •Featured in the San Francisco Chronicle, "California just invested millions in lab-grown meat, becoming the first state to back the unproven industry" (L. Zimberoff, Jul 2022)? Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project has provided the following training and professional development opportunities: •One-on-one research mentorship of PI Rowat with trainees Sam Norris, Corinne Smith, Stephanie Kawecki, Kathleen Chen, and Qingwen Xie. • Graduate student researcher, Stephanie Kawecki, presented a talk and organized a feedback sessionwith advisors at Michigan State University. • Graduate student researcher, Kathleen Chen presented a poster on 'Scalable Production of Adipose Tissue Using Edible Microcarrier Scaffolds with Tunable Stiffness'at the Gordon Research Conference on "Convergence of Nanotechnology with Food and Agriculture", Manchester, NH, June 2022. •Graduate student researcher, Stephanie Kawecki, presented a poster on 'Cultured Meat with Tunable Fat Content: Scaffolds for Multicomponent Tissue with Myocytes and Adipocytes' at the Society for Biomaterials, Baltimore, MD, April 2022. •Mentorship provided by PI Rowat for graduate and undergraduate students to lead an Alt Protein Project initiative at UCLA. How have the results been disseminated to communities of interest?Resultshave been disseminated through invited talks that the PI presented during the award period: PEER-REVIEWED PUBLICATIONS: •Norris SCP,Davis AR,Kawecki NS,Chen KK,Rowat AC#(2022) Emulsion-templated microparticles with tunable stiffness and topology: applications as edible microcarriers for cultured meat.Biomaterials.287: 121699 •Kawecki NS,Rowat AC#(2023) Future Pathways in Biosynthesis: Livestock Tissue Culture in 'The Protein Transition'. Eds. Pyett S, van Mierlo B, Trindade L, de Vet E, Welch D, van Zanten H, de Jong L, and van den Top M [Invited Book Chapter] •Kawecki NS,Norris SCP, Xu Y,Davis AR,Fridman E,Chen KK, Garmyn A, Mason T,Rowat AC#.Towards marbled cultured meat: Engineering multicomponent tissues using scaffolds with customized physical properties to support myogenesis and adipogenesis.Revisions requested. INVITED LECTURES BY PI ROWAT: International Teaching Kitchen Research Conference, Los Angeles, USA (Panel, 'Breaking Departmental Silos and Forging New Interdisciplinary Partnerships to Innovate in Research, Academics, and Operations') Western Association of Core Directors Annual Meeting, UCLA Michigan State University, G. Malcolm Trout Visiting Scholar Program 33rdAnnual Lecture, Department of Food Science & Human Nutrition Biophysical Society Annual Meeting, San Francisco, USA (Symposium,'Mechanosensation') Geilo 50thWinter School, Geilo, Norway UCLA Women in Philanthropy Meeting (*Included cooking demonstration of beef vs. plant-based burgers and discussion of factors that impact food choices) UCLA Division of Life Sciences Advisory Board Meeting UCLA Department of Integrative Biology & Physiology Chair's Committee Roundtable EVENTS FOR GENERAL AUDIENCES: PI Rowat spearheaded and led the public event, 'People, Food, & Climate', hosted by Science&Food and the Los Angeles Times Food Bowl (Sept 2022). UNDERGRADUATE AND HIGH SCHOOL EDUCATION: PI Rowat was an invited speaker for the 5thAnnual UCLA at Geffen Academy Day, Los Angeles (March 2022) PI Rowat was an invited speaker for the University of North Carolina Bio290 "The Cellular Agriculture Revolution" (March 2022) PI Rowat discussed cultured meat in the interdisciplinary undergraduate class that she developed and taught in spring 2022, FOOD STUDIES 181,Special Topics: Perspectives on Food and Society, Spring 2022. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
There is urgent need to diversify methods for protein production so that our future food system will be resilient to disruptions due to the impacts of climate variability, natural disasters, and future epidemics.One potential emerging approach uses tissue culture methods to enable cultured meat. The vision of this project is to develop cultured meat with properties that can be tailored for defined flavor and nutritional properties, from a lean, low-cholesterol filet mignon to a marbled rib-eye rich in omega-3 fatty acids. Critical for this vision are physical scaffolds that mimic the natural growth environment of muscle and fat cells. Scaffolds impact cell behaviors critical for the efficient production of palatable cultured meat: the stiffness of scaffolds can regulate cell growth, and also the quality of muscle and fat tissue, which is important for meat sensory and nutritional properties, including texture and mouthfeel. The goal of this specific project is to produce a cultured meat burger with defined fat content to meet different consumer needs, and to identify supplements that enable us to tune the lipid composition of cultured meat towards achieving nutritious and flavorful cultured meat.More broadly, our findings should contribute to the development of cultured meat as a feasible method for protein production, which can ultimately support the increased resiliency of future food systems. In the annual reporting period, we have made strong progress towards our goal of producing a cultured meat burger with defined nutrition and sensory properties to meet consumer needs.Our strategyis to develop edible 'microcarrier' scaffolds that support the growth of cultured tissues in a large vessel or bioreactor, which has potential to scale up tissue culture that is required for food production. We have: 1) Developed edible microcarrier scaffolds to support cultured muscle microtissue (Objective 1; Norris et alBiomaterials2022); 2) Developed edible microcarrier scaffolds to support cultured fat microtissue (Objective 2; Kawecki et al, Revisions requested); 3) Demonstrated that edible microcarrier scaffolds can be used to generate proof of concept cultured ground beef, which is cookable and shows browning characteristic of Maillard reactions (Objective 3; Norris et alBiomaterials2022).In addition, negotiations are in progress for licensing our patent on 'Methods for Producing Cultured Meat that has Heterogeneous Composition'to the startup company, EarthFlavr. Objective 1: Develop edible microcarrier scaffolds to support cultured muscle microtissue. Major activities completed:To fabricate edible microcarriers, we usewater-in-oil emulsions as templates for gelatin microparticles. Based on previous findings that striated substrates promote myoblast proliferation and myotube formation, we developed an embossing method to imprint grooved surface topology on edible microcarriers.We analyzed the proliferation of mouse myogenic C2C12 cells on edible microcarriers by measuringgenomic DNA.To confirm differentiation, we visualized the myogenic marker myosin heavy chain 4 (Myh4) in microtissues by confocal microscopy. Data collected: We found that cells were attached to all types of microcarriers at days 1, 4, and 7 using widefield and confocal microscopy. For smooth and grooved microcarriers (sMCs and gMCs respectively), the cell density was significantly higher after 8 days of culture (p = 5.5 x10-4 for sMCs and p = 6.9 x10-5 for gMCs, d0 vs. d8 one-way ANOVA), showing that edible microcarriers supported cell proliferation. Consistent with skeletal muscle formation, we observed increased fluorescence intensity of immunolabeledMyh4, indicating increased expression at the protein level. Summary and discussion of results:In this proof-of-concept demonstration, we showed that edible microcarriers with both smooth and grooved surface topologies supported the proliferation and differentiation of mouse myogenic C2C12 cells in a suspension culture. The grooved edible microcarriers showed a modest increase in the proliferation and alignment of myogenic cells compared to cells cultured on smooth, spherical microcarriers. Key outcomes:1)Edible microcarriers can support the generation of bovine and murine myogenic microtissues in suspension culture. 2)Results show the potential of using edible microcarriers with tunable stiffness and topology to reduce the volume fraction of scaffolds in the final cultured meat product. Objective 2: Develop edible microcarrier scaffolds to support cultured fat microtissue with desired lipid content. Major activities completed: We designed microbead scaffolds withE~4 kPa, which mimics the stiffness of adipose tissue. Pre-adipocytes are proliferated and differentiated on microbead scaffolds.We measure the accumulation of lipids in3T3-L1 pre-adipocytes on microbeads.We also assess if adipogenic microtissues can adhere with myogenic microtissues into a mechanically stable multicomponent tissue using a parallel plate rheometer. Data collected:We confirm the Young's modulus of the gelatin microbeads isE~ 4.4 ± 0.51 kPa using AFM. Over the 14 day time course, we observe increased accumulation of intracellular lipids in both rabbit and mouse adipocytes, as evidenced by the increased signal of LipidTox. The 3T3-L1 adipocytes show a ~100xincrease in lipid area per cell over the 14 day timescale as demonstrated by quantitative image analysis. We find a similar trend of lipid accumulation in the rabbit RbPreAd cells--albeit with only a ~20x increase. Summary and discussion of results:Experiments using the parallel plate rheometerreveal that the tissue resists increasing magnitudes of applied strain, indicating the sample exhibits properties of a solid material.Future studies will decipher the effects of microbead stiffness in a 3D context on adipose microtissue growth. Key outcomes:1)Engineering of scaffolds with customized physical properties that provide the foundation for structuring and organizing both myogenic and adipogenic cells as well as microtissues. 2)Multicomponent 3D tissue constructs self-assemble without the use of additional crosslinkers, which can reduce the number of post-processing steps and required reagents. Objective 3: Produce cultured ground beef with desired qualities using edible microcarrier scaffolds. Major activities completed:To investigate the ability of the edible microcarriers to support the generation of cultured meat, we cultured bovine satellite muscle cells (BSMCs)on edible microcarriersin a spinner flask. We harvest the microtissues by centrifugation toform a consolidated cultured meat patty andevaluate cookability of the cultured meat. Data collected:Upon heating, the cultured meat patty retained its shape; by contrast, the control gelatin samples melted and lost their form. The cultured meat also exhibited browning, characteristic of Maillard reactions. Summary and discussion of results: 1)Edible microcarriers supported the production of myogenic microtissue from C2C12 orBSMCs, which we harvested by centrifugation into a cookable meat patty that maintained its shape and exhibited browning during cooking. 2)Findings show the potential of edible microcarriers for the scalable production of cultured meat in a single bioreactor. Key outcomes:The scalable approach that we describe to generate edible microcarriers and the resultant muscle microtissues has potential to contribute to efficient, cost-effective cultured meat production, which could provide a complementary alternative for protein production that ultimately could help to increase the resilience of future food systems.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Norris SCP, Davis AR, Kawecki NS, Chen KK, Rowat AC (2022) Emulsion-templated microparticles with tunable stiffness and topology: applications as edible microcarriers for cultured meat. Biomaterials. 287: 121699
- Type:
Book Chapters
Status:
Awaiting Publication
Year Published:
2023
Citation:
Kawecki NS, Rowat AC (2023) Future Pathways in Biosynthesis: Livestock Tissue Culture in The Protein Transition. Eds. Pyett S, van Mierlo B, Trindade L, de Vet E, Welch D, van Zanten H, de Jong L, and van den Top M [Invited Book Chapter]
- Type:
Journal Articles
Status:
Under Review
Year Published:
2023
Citation:
Kawecki NS, Norris SCP, Xu Y, Davis AR, Fridman E, Chen KK, Garmyn A, Mason T, Rowat AC. Towards marbled cultured meat: Engineering multicomponent tissues using scaffolds with customized physical properties to support myogenesis and adipogenesis. Revisions requested.
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