Аннотация
A quarter of the world’s population has no access to safe foods of high quality due to the inability of traditional agriculture to meet the growing needs. Therefore, cultivated meat produced from a large mass of animal cells in a laboratory is becoming a promising alternative to animal products. In this study, we aimed to develop a technology for obtaining a hybrid cultured meat product from rabbit cells, sodium alginate, and sunflower protein, as well as to analyze its morphological and functional characteristics.We used rabbit stem cells isolated from the greater omentum and exposed to lipogenic and myogenic differentiation, as well as rabbit skin fibroblasts. The cells were placed in a hydrogel of sodium alginate and sunflower protein and cultured for 72 h to biofabricate tissue constructs by using 3D bioprinting. Confocal and transmission electron microscopy was applied to analyze the morphological and functional characteristics of the cells in the constructs.
Using 3D bioprinting, we obtained tissue constructs of 30×40×3 mm from rabbit cells, sodium alginate, and sunflower protein. According to confocal microscopy, the cells in the tissue constructs remained viable for at least 72 h. Transmission electron microscopy showed that the cells formed tight junctions and were metabolically active for at least 72 h, with fibroblasts secreting procollagen and lipoblasts secreting lipid droplets.
The resulting cellular meat was obtained from a combination of fibroblasts, lipocytes, and myogenic cells, as well as two ink components. The cellular meat product was safe and ready for consumption.
Ключевые слова
Cultured meat, tissue construct, 3D bioprinting, stem cells, rabbit, vegetable proteinСПИСОК ЛИТЕРАТУРЫ
- Pison G, Couppié E, Caporali A. Tous les pays du monde. Population and Sociétés. 2022;(603):1–8. https://doi.org/10.3917/popsoc.603.0001
- Sustainable Development Goals. Goal 2: Zero Hunger [Internet]. [cited 2024 Mar 20]. Available from: https://www.un.org/sustainabledevelopment/hunger/
- de Castro Cardoso Pereira PM, dos Reis Baltazar Vicente AF. Meat nutritional composition and nutritive role in the human diet. Meat Science. 2013;93(3):586–592. https://doi.org/10.1016/j.meatsci.2012.09.018
- Groombridge B, Jenkins M. World Atlas of Biodiversity: Earth's Living Resources in the 21st Century. California: University of California Press; 2002. 340 p.
- Broom DM. Animal welfare complementing or conflicting with other sustainability issues. Applied Animal Behaviour Science. 2019;219:104829. https://doi.org/10.1016/j.applanim.2019.06.010
- Losada-Espinosa N, Miranda-De la Lama GC, Estévez-Moreno LX. Stockpeople and Animal Welfare: Compatibilities, Contradictions, and Unresolved Ethical Dilemmas. Journal of Agricultural and Environmental Ethics. 2020;33:71–92. https://doi.org/10.1007/s10806-019-09813-z
- Stavins R, Wagner AF, Wagner G. Interpreting Sustainability in Economic Terms: Dynamic Efficiency Plus Intergenerational Equity. Economics Letters. 2003;79(3):339–343. https://doi.org/10.1016/S0165-1765(03)00036-3
- Sha L, Xiong YL. Plant protein-based alternatives of reconstructed meat: Science, technology, and challenges. Trends in Food Science and Technology. 2020;102:51–61. https://doi.org/10.1016/j.tifs.2020.05.022
- Koyande AK, Chew KW, Rambabu K, Tao Y, Chu DT, Show PL. Microalgae: A potential alternative to health supplementation for humans. Food Science and Human Wellness. 2019;8(1):16–24. https://doi.org/10.1016/j.fshw.2019.03.001
- Liceaga AM, Aguilar-Toalá JE, Vallejo-Cordoba B, González-Córdova AF, Hernández-Mendoza A. Insects as an alternative protein source. Annual Review of Food Science and Technology. 2022;13:19–34. https://doi.org/10.1146/annurev-food-052720-112443
- Molfetta M, Morais EG, Barreira L, Bruno GL, Porcelli F, Dugat-Bony E, et al. Protein sources alternative to meat: state of the art and involvement of fermentation. Foods. 2022;11(14):2065. https://doi.org/10.3390/foods11142065
- Macdiarmid J. The food system and climate change: Are plant-based diets becoming unhealthy and less environmentally sustainable? Proceedings of the Nutrition Society. 2022;81(2):162–167. https://doi.org/10.1017/S0029665121003712
- Siegrist M, Hartmann C. Why alternative proteins will not disrupt the meat industry. Meat Science. 2023;203:109223. https://doi.org/10.1016/j.meatsci.2023.109223
- Onwezen MC, Bouwman EP, Reinders MJ, Dagevos H. A systematic review on consumer acceptance of alternative proteins: Pulses, algae, insects, plant-based meat alternatives, and cultured meat. Appetite. 2020;159:105058. https://doi.org/10.1016/j.appet.2020.105058
- Post MJ. Cultured meat from stem cells: Challenges and prospects. Meat Science. 2012;92(3):297–301. https://doi.org/10.1016/j.meatsci.2012.04.008
- Post MJ, Levenberg S, Kaplan DL, Genovese N, Fu J, Bryant CJ, et al. Scientific, sustainability and regulatory challenges of cultured meat. Nature Food. 2020;1:403–415. https://doi.org/10.1038/s43016-020-0112-z
- Chen L, Guttieres D, Koenigsberg A, Barone PW, Sinskey AJ, Springs SL. Large-scale cultured meat production: Trends, challenges and promising biomanufacturing technologies. Biomaterials. 2022;280:121274. https://doi.org/10.1016/j.biomaterials.2021.121274
- Ganesh KS, Sridhar A, Vishali S. Utilization of fruit and vegetable waste to produce value-added products: Conventional utilization and emerging opportunities-A review. Chemosphere. 2022;287(3):132221. https://doi.org/10.1016/j.chemosphere.2021.132221
- Tassoni A, Tedeschi T, Zurlini C, Cigognini IM, Petrusan J-I, Rodríguez Ó, et al. State-of-the-Art Production Chains for Peas, Beans and Chickpeas–Valorization of Agro-Industrial Residues and Applications of Derived Extracts. Molecules. 2020;25(6):1383. https://doi.org/10.3390/molecules25061383
- de Oliveira Filho JG, Egea MB. Sunflower seed byproduct and its fractions for food application: An attempt to improve the sustainability of the oil process. Journal of Food Science. 2021;86(5):1497–1510. https://doi.org/10.1111/1750-3841.15719
- Nezlek JB, Forestell CA. Meat substitutes: current status, potential benefits, and remaining challenges. Current Opinion in Food Science. 2022;47:100890. https://doi.org/10.1016/j.cofs.2022.100890
- Grasso S, Asioli D, Smith R. Consumer co-creation of hybrid meat products: A cross-country European survey. Food Quality and Preference. 2022;100:104586. https://doi.org/10.1016/j.foodqual.2022.104586
- Ovsianikov A, Yoo J, Mironov V. 3D printing and biofabrication. ACM Digital Library: Springer Publishing Company; 2018. 563 p.
- Dey M, Ozbolat IT. 3D bioprinting of cells, tissues and organs. Scientific Reports. 2020;10:14023. https://doi.org/10.1038/s41598-020-70086-y
- Nachal N, Moses JA, Karthik P, Anandharamakrishnan C. Applications of 3D Printing in Food Processing. Food Engineering Reviews. 2019;11:123–141. https://doi.org/10.1007/s12393-019-09199-8
- Seo J W, Jung WK, Park YH, Bae H. Development of cultivable alginate fibers for an ideal cell-cultivated meat scaffold and production of hybrid cultured meat. Carbohydrate Polymers. 2023;321:121287. https://doi.org/10.1016/j.carbpol.2023.121287
- Chen Y, Zhang M, Bhandari B. 3D Printing of Steak-like Foods Based on Textured Soybean Protein. Foods. 2021;10(9):2011. https://doi.org/10.3390/foods10092011
- Srutee R, Sowmya RS, Annapure US. Clean meat: techniques for meat production and its upcoming challenges. Animal Biotechnology. 2022;33(7):1721–1729. https://doi.org/10.1080/10495398.2021.1911810
- Bunnell BA, Flaat M, Gagliardi C, Patel B, Ripoll C. Adipose-derived stem cells: isolation, expansion and differentiation. Methods. 2008;45(2):115–120. https://doi.org/10.1016/j.ymeth.2008.03.006
- Lund P, Pilgaard L, Duroux M, Fink T, Zachar V. Effect of growth media and serum replacements on the proliferation and differentiation of adipose-derived stem cells. Cytotherapy. 2009;11(2):189–197. https://doi.org/10.1080/14653240902736266
- Meligy FY, Shigemura K, Behnsawy HM, Fujisawa M, Kawabata M, Shirakawa T. The efficiency of in vitro isolation and myogenic differentiation of MSCs derived from adipose connective tissue, bone marrow, and skeletal muscle tissue. In Vitro Cellular and Developmental Biology – Animal. 2012;48:203–215. https://doi.org/10.1007/s11626-012-9488-x
- Freshney RI. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications. Hoboken, New Jersey: John Wiley and Sons; 2015. pp.383–401
- Abade dos Santos FA, Carvalho CL, Almeida I, Fagulha T, Rammos F, Barros SC, et al. Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures. Cells. 2021;10(8):2100. https://doi.org/10.3390/cells10082100
- Regenstein JM, Chaudry MM, Regenstein CE. The kosher and halal food laws. Comprehensive reviews in food science and food safety. 2003;2(3):111–127. https://doi.org/10.1111/j.1541-4337.2003.tb00018.x
- Kenigsberg JA, Zivotofsky AZ. A Jewish Religious Perspective on Cellular Agriculture. Frontiers in Sustainable. Food Systems. 2019;3:128. https://doi.org/10.3389/fsufs.2019.00128
- Farouk MM, Regenstein JM, Pirie MR, Najm R, Bekhit AED, Knowles SO. Spiritual aspects of meat and nutritional security: Perspectives and responsibilities of the Abrahamic faiths. Food Research International. 2015;76: 882–895. https://doi.org/10.1016/j.foodres.2015.05.028
- Axpe E, Oyen ML. Applications of alginate-based bioinks in 3D bioprinting. International Journal of Molecular Sciences. 2016;17(12):1976. https://doi.org/10.3390/ijms17121976
- Gao Q, Kim BS, Gao G. Advanced strategies for 3D bioprinting of tissue and organ analogs using alginate hydrogel bioinks. Marine Drugs. 2021;19(12):708. https://doi.org/10.3390/md19120708
- Qin Y, Jiang J, Zhao L, Zhang J, Wang F. Applications of Alginate as a Functional Food Ingredient. In: Grumezescu AM, Holban AM. Biopolymers for Food Design. Academic Press; 2018. pp. 409–429. https://doi.org/10.1016/b978-0-12-811449-0.00013-x
- Puscaselu GR, Lobiuc A, Dimian M, Covasa M. Alginate: From Food Industry to Biomedical Applications and Management of Metabolic Disorders. Polymers. 2020;12(10):2417. https://doi.org/10.3390/polym12102417
- Joo ST, Choi JS, Hur SJ, Kim GD, Kim CJ, Lee EY, et al. A comparative study on the taste characteristics of satellite cell cultured meat derived from chicken and cattle muscles. Food Science of Animal Resources. 2022;42(1):175–185. https://doi.org/10.5851/kosfa.2021.e72
- Tomasevic I, Djekic I, Font-i-Furnols M, Terjung N, Lorenzo JM. Recent advances in meat color research. Current Opinion in Food Science. 2021;41:81–87. https://doi.org/10.1016/j.cofs.2021.02.012
- Rasband W. ImageJ User Guide. (1997–2018). National Institutes of Health, Bethesda, Maryland, USA. https://imagej.net/ij/
- Santos ACA, Camarena DEM, Roncoli Reigado G, Chambergo FS, Nunes VA, Trindade MA, et al. Tissue Engineering Challenges for Cultivated Meat to Meet the Real Demand of a Global Market. International Journal of Molecular Sciences. 2023;24(7):6033. https://doi.org/10.3390/ijms24076033
- Battle M, Bomkamp C, Carter M, Colley Clarke J, Fathman L, Gertner D, et al. 2023 State of the industry report: Cultivated meat and seafood. Washington DC: The Good Food Institute; 2023. 74 p. https://gfi.org/wp-content/uploads/2024/04/2023-State-of-the-Industry-Report-Cultivated-meat-and-seafood.pdf