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Cultivated meat (CM) refers to edible lab-grown meat that incorporates cultivated animal cells. It has the potential to address some issues associated with real meat (RM) production, including the ethical and environmental impact of animal farming, and health concerns. Recently, various biomanufacturing methods have been developed to attempt to recreate realistic meat in the laboratory. We therefore overview recent achievements and challenges in the production of CM. We also discuss the issues that need to be addressed and suggest additional recommendations and potential criteria to help to bridge the gap between CM and RM from an engineering standpoint., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Cultivated meat (CM) has transitioned from a futuristic concept to a present reality, with select products approved for consumption and sale in Singapore, Israel, and the USA. This evolution has emphasized scalable, cost-effective, and sustainable production, as well as navigation of regulatory pathways. As CM develops, a crucial challenge lies in delivering products that are highly appealing to consumers. Central to this will be refining CM palatability, a term encompassing food's taste, aroma, texture, tenderness, juiciness, and color. We explore the scientific and engineering approaches to producing palatable CM, including cell-line selection, cell differentiation, and post-processing techniques. This includes a discussion of the structural and compositional properties of meat that are intrinsically coupled to palatability., Competing Interests: Declaration of interests This work was produced as a collaboration between academia and industry. C.J.O is an employee of Smart MCs Pty Ltd, a company developing biomaterials for cell culture applications, including cultivated meat. A.O.W. is a founding employee of Arta Bioanalytics, an Australian company working in bioinformatics for cultivated meat. Contributions by C.J.O and A.O.W were also undertaken during their employment at Vow Group Pty Ltd., an Australian company focused on the development of cultivated meat. The views and scientific conclusions expressed in this manuscript are those of the authors and do not represent the views of their current or former affiliated organizations. Aside from the aforementioned, the other authors declare that they have no competing interests influencing this work., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The emergence of cultivated meat provides a sustainable and ethical alternative to traditional animal agriculture, highlighting its increasing importance in the food industry. Biomaterial scaffolds are critical components in cultivated meat production for enabling cell adhesion, proliferation, differentiation, and orientation. While there's extensive research on scaffolding biomaterials, applying them to cultivated meat production poses distinct challenges, with each material offering its own set of advantages and disadvantages. This review summarizes the most recent scaffolding biomaterials used in the last five years for cell-cultured meat, detailing their respective advantages and disadvantages. We suggest future research directions and provide recommendations for scaffolds that support scalable, cost-effective, and safe high-quality meat production. Additionally, we highlight commercial challenges cultivated meat faces, encompassing bioreactor design, cell culture mediums, and regulatory and food safety issues. In summary, this review provides a comprehensive guide and valuable insights for researchers and companies in the field of cultivated meat production., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Shue Wang reports financial support was provided by National Science Foundation (CMMI: 2143151). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cultivated meat, an advancement in cellular agriculture, holds promise in addressing environmental, ethical, and health challenges associated with traditional meat production. Utilizing tissue engineering principles, cultivated meat production employs biomaterials and technologies to create cell-based structures by introducing cells into a biocompatible scaffold, mimicking tissue organization. Among the cell sources used for producing muscle-like tissue for cultivated meats, primary adult stem cells like muscle satellite cells exhibit robust capabilities for proliferation and differentiation into myocytes, presenting a promising avenue for cultivated meat production. Evolutionarily optimized for growth in a 3D microenvironment, these cells benefit from the biochemical and biophysical cues provided by the extracellular matrix (ECM), regulating cell organization, interactions, and behavior. While plant protein-based scaffolds have been explored for their utilization for cultivated meat, they lack the biological cues for animal cells unless functionalized. Conversely, a decellularized bovine placental tissue ECM, processed from discarded birth tissue, achieves the biological functionalities of animal tissue ECM without harming animals. In this study, collagen and total ECM were prepared from decellularized bovine placental tissues. The collagen content was determined to be approximately 70% and 40% in isolated collagen and ECM, respectively. The resulting porous scaffolds, crosslinked through a dehydrothermal (DHT) crosslinking method without chemical crosslinking agents, supported the growth of bovine myoblasts. ECM scaffolds exhibited superior compatibility and stability compared to collagen scaffolds. In an attempt to make cultivate meat constructs, bovine myoblasts were cultured in steak-shaped ECM scaffolds for about 50 days. The resulting construct not only resembled muscle tissues but also displayed high cellularity with indications of myogenic differentiation. Furthermore, the meat constructs were cookable and able to sustain the grilling/frying. Our study is the first to utilize a unique bovine placentome-derived ECM scaffold to create a muscle tissue-like meat construct, demonstrating a promising and sustainable option for cultivated meat production.

In recent years, there has been widespread adoption of machine learning (ML) technologies to unravel intricate relationships among diverse parameters in various additive manufacturing (AM) techniques. These ML models excel at recognizing complex patterns from extensive, well-curated datasets, thereby unveiling latent knowledge crucial for informed decision-making during the AM process. The collaborative synergy between ML and AM holds the potential to revolutionize the design and production of AM-printed parts. This review delves into the challenges and opportunities emerging at the intersection of these two dynamic fields. It provides a comprehensive analysis of the publication landscape for ML-related research in the field of AM, explores common ML applications in AM research (such as quality control, process optimization, design optimization, microstructure analysis, and material formulation), and concludes by presenting an outlook that underscores the utilization of advanced ML models, the development of emerging sensors, and ML applications in emerging AM-related fields. Notably, ML has garnered increased attention in AM due to its superior performance across various AM-related applications. It is envisioned that the integration of ML into AM processes will significantly enhance 3D printing capabilities across diverse AM-related research areas., (© 2024 The Authors. Advanced Materials published by Wiley‐VCH GmbH.)

Cultivated meat, also known as cell-based or clean meat, utilizes mesenchymal stem cells to cultivate mature cell types like adipocytes, which are pivotal for imparting the desired taste and texture. The delivery of differentiated cells, crucial in cultivated meat production, is facilitated through extensive exploration of 3D culturing techniques mimicking physiological environments. In this study, we investigated the adipogenic differentiation potential of bovine umbilical cord stem cells (BUSCs), sourced from discarded birth tissue, and assessed the feasibility of delivering differentiated cells for cultivated meat using gelatin methacrylate (GelMA) as a carrier for adipose tissue. Various adipogenic inducers, previously reported to be effective for human mesenchymal stem cells (hMSCs), were evaluated individually or in combination for their efficacy in promoting the adipogenesis of BUSCs. Surprisingly, while the traditional adipogenic inducers, including insulin, dexamethasone, isobutylmethylxantine (IBMX), indomethacin, and rosiglitazone, showed no significant effect on the adipogenic differentiation of BUSCs, efficient differentiation was achieved in the presence of a fatty acid cocktail. Furthermore, we explored methods for the delivery of BUSCs. Differentiated cells were delivered either encapsulated in GelMA hydrogel or populated on the surface of GelMA microparticles (MPs) as the adipose component of cultivated meat. Our findings reveal that after adipogenic induction, the lipid production per cell was comparable when cultured either within hydrogel or on MPs. However, GelMA-MPs supported better cell growth compared to hydrogel encapsulation. Consequently, the overall lipid production is higher when BUSCs are delivered via GelMA-MPs rather than encapsulation. This study not only systematically evaluated the impact of common adipogenic inducers on BUSCs, but also identified GelMA-MPs as a promising carrier for delivering bovine adipocytes for cultivated meat production.

There is increasing consumer demand for alternative animal protein products that are delicious and sustainably produced to address concerns about the impacts of mass-produced meat on human and planetary health. Cultured meat has the potential to provide a source of nutritious dietary protein that both is palatable and has reduced environmental impact. However, strategies to support the production of cultured meats at the scale required for food consumption will be critical. In this review, we discuss the current challenges and opportunities of using edible scaffolds for scaling up the production of cultured meat. We provide an overview of different types of edible scaffolds, scaffold fabrication techniques, and common scaffold materials. Finally, we highlight potential advantages of using edible scaffolds to advance cultured meat production by accelerating cell growth and differentiation, providing structure to build complex 3D tissues, and enhancing the nutritional and sensory properties of cultured meat.

Cultured meat (also referred to as cultivated, cell-based, or cell-cultured meat) is a novel food technology that is presented as a method of meat production without reliance on large-scale industrial farming. The pro-cultured meat narrative rests, in part, on a moral foundation: cultured meat is purported to alleviate the environmental and animal welfare harms associated with farmed meat. Despite this narrative, no research has examined which moral values underpin attitudes towards cultured meat. To examine this, we surveyed 1861 participants from the United States and Germany about their moral foundations and their attitudes towards cultured meat. In line with predictions, people who more strongly endorse moral values about purity (i.e., had higher scores on the purity subscale of the moral foundations scale) held more negative attitudes towards cultured meat. However, this relationship was much more consistent among participants from the United States than participants from Germany. Against predictions, attitudes towards cultured meat were not reliably associated with the extent to which people focus on harm as a moral foundation. The latter finding was particularly surprising in light of harm-reduction narratives around cultured meat. These findings demonstrate the need for a more nuanced discussion about, and understanding of, consumer concerns around cultured meat and the values that underpin them., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Growth factors are commonly added to cell culture media in cellular agriculture to mimic the endogenous process of proliferation and differentiation of cells. Many of these growth factors are endogenous to humans and known to be present in the edible tissues and milk of food animals. However, there is little or no information on the use of growth factors intentionally added in food production before the advent of cultivated meat. Ten commonly used growth factors have been reviewed to include information on their mode of action, bioavailability, occurrence in food and food animals, endogenous levels in humans, as well as exposure and toxicological information drawn from relevant animal studies and human clinical trials with a focus on oral exposure. In addition, a comparison of homology of growth factors was done to compare the sequence homology of growth factors from humans and domestic animal species commonly consumed as food, such as bovine, porcine, and poultry. This information has been gathered as the starting point to determine the safety of use of growth factors in cultivated meat meant for human consumption. The change in levels of growth factors measured in human milk and bovine milk after pasteurization and high-temperature treatment is discussed to give an indication of how commercial food processing can affect the levels of growth factors in food. The concept of substantial equivalence is also discussed together with a conservative exposure estimation. More work on how to integrate in silico assessments into the routine safety assessment of growth factors is needed., (© 2024 Institute of Food Technologists®.)

Modern food technology has given rise to numerous alternative protein sources in response to a growing human population and the negative environmental impacts of current food systems. To aid in achieving global food security, one such form of alternative protein being investigated is cultivated meat, which applies the principles of mechanical and tissue engineering to produce animal proteins and meat products from animal cells. Herein, nonmodified and methacrylated whey protein formed hydrogels with methacrylated alginate as potential tissue engineering scaffolds for cultivated meat. Whey protein is a byproduct of dairy processing and was selected because it is an approved food additive and cytocompatible and has shown efficacy in other biomaterial applications. Whey protein and alginate scaffolds were formed via visible light cross-linking in aqueous solutions under ambient conditions. The characteristics of the precursor solution and the physical-mechanical properties of the scaffolds were quantified; while gelation occurred within the homo- and copolymer hydrogels, the integrity of the network was significantly altered with varying components. Qualitatively, the scaffolds exhibited a three-dimensional (3D) interconnected porous network. Whey protein isolate (WPI)-based scaffolds were noncytotoxic and supported in vitro myoblast adhesion and proliferation. The data presented support the hypothesis that the composition of the hydrogel plays a significant role in the scaffold's performance.

Since the introduction of the cultivated meat burger in 2013, numerous discussions have transpired between researchers and consumers, manifesting in various forms such as academic publications, opinions on personal websites and interactions on social platforms. For Muslim consumers, a primary concern revolves around the halal status of cultivated meat, given the pivotal role of adhering to a halal diet as a divine obligation in their lives. Therefore, this article seeks to systematically review the existing literature on cultivated meat from an Islamic perspective as articulated by Muslim scholars, whether these perspectives are issued by an established fatwa organisation or representations of personal views. The sources incorporated into this analysis span from academic publications, newspaper articles, fatwa bodies, personal websites and interviews. Employing thematic analysis, five principal themes were discerned within the discourse among Muslim scholars regarding cultivated meat: (i) the ontological status of cultivated meat, (ii) the status and source of stem cells used in the cultivation process, (iii) the theological implications of cultivated meat production on altering God's creation, (iv) foundational principles for halal cultivated meat production, and (v) the contemporary necessity of cultivated meat from the perspective of al-maqasid al-shariah . It is duly recommended that international fatwa organisations such as the International Islamic Fiqh Academy (IIFA) engage in discussions and deliberations on this matter of growing significance within the food industry. The fatwas and resolutions issued by IIFA are frequently cited as the most recognised authority in many Islamic countries., Competing Interests: Mohammad Naqib Hamdan reports financial support was provided by 10.13039/501100005417Universiti Teknologi Malaysia., (© 2024 The Authors.)

Cultivated meat is emerging to replace traditional livestock industries, which have ecological costs, including land and water overuse and considerable carbon emissions. During cultivated meat production, mammalian cells can increase their numbers dramatically through self-renewal/proliferation and transform into mature cells, such as muscle or fat cells, through maturation/differentiation. Here, we address opportunities for introducing probiotic bacteria into the cultivated meat industry, including using them to produce renewable antimicrobials and scaffolding materials. We also offer solutions to challenges, including the growth of bacteria and mammalian cells, the effect of probiotic bacteria on production costs, and the effect of bacteria and their products on texture and taste. Our summary provides a promising framework for applying microbial composites in the cultivated meat industry., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Cultivated meat production is a promising technology to generate meat while reducing the reliance on traditional animal farming. Biomaterial scaffolds are critical components in cultivated meat production, enabling cell adhesion, proliferation, differentiation, and orientation. In the present work, naturally derived glutenin was fabricated into films with and without surface patterning and in the absence of toxic cross-linking or stabilizing agents for cell culture related to cultivated meat goals. The films were stable in culture media for at least 28 days, and the surface patterns induced cell alignment and guided myoblast organization (C2C12s) and served as a substrate for 3T3-L1 adipose cells. The films supported adhesion, proliferation, and differentiation with mass balance considerations (films, cells, and matrix production). Freeze-thaw cycles were applied to remove cells from glutenin films and monitor changes in glutenin mass with respect to culture duration. Extracellular matrix (ECM) extraction was utilized to quantify matrix deposition and changes in the original biomaterial mass over time during cell cultivation. Glutenin films with C2C12s showed mass increases with time due to cell growth and new collagen-based ECM expression during proliferation and differentiation. All mass balances were compared among cell and noncell systems as controls, along with gelatin control films, with time-dependent changes in the relative content of film, matrix deposition, and cell biomass. These data provide a foundation for cell/biomaterial/matrix ratios related to time in culture as well as nutritional and textural features.

Cultivated meat is an emerging new technology to produce sustainable meat for the future. The common approach for cultivated meat, is the isolation of satellite cells from donor animals, followed by in vitro proliferation and differentiation into primitive muscle fibers. The transformation of satellite cells into myofibers is tightly orchestrated by intra-cellular signaling, while the inter-cellular signaling is less well understood. Thus, the current study was conducted to map the secretion of potential signaling molecules (MicroRNAs and proteins) during proliferation and differentiation. Primary cultures of satellite cells were grown to 50% and 80% confluence, representing the proliferative phase or serum-starved for 1 and 3 days to induce differentiation. Post incubation in FBS-free media, the media were collected and analyzed for miRNA and protein content using gene-arrays and LC-MS/MS, respectively. When comparing the miRNA secretome at 50% and 80% confluence, we observed four differentially expressed miRNA, while only five were differentially expressed when comparing Day 1 to Day 3. A subsequent in silico analysis suggested that pathways of importance for myogenesis, e.g., MAPK and AMPK signaling, could be regulated by the secreted miRNAs. In addition, >300 proteins were secreted, including insulin-like growth factor 1 binding proteins 2, 3, 4, 5 and 6. In conclusion, this study demonstrated differential secretion of several miRNAs and proteins during both proliferation and differentiation of bovine satellite cells in vitro., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jette Feveile Young reports financial support was provided by The Good Food Institute., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Overcoming the challenge of creating thick, tissue-resembling muscle constructs is paramount in the field of cultivated meat production. This study investigates the remarkable potential of random cellulose acetate nanofibers (CAN) as a transformative scaffold for muscle tissue engineering (MTE), specifically in the context of cultivated meat applications. Through a comparative analysis between random and aligned CAN, utilizing C2C12 and H9c2 myoblasts, we unveil the unparalleled capabilities of random CAN in facilitating muscle differentiation, independent of differentiation media, by exploiting the YAP/TAZ-related mechanotransduction pathway. In addition, we have successfully developed a novel process for stacking cell-loaded CAN sheets, enabling the production of a three-dimensional meat product. C2C12 and H9c2 loaded CAN sheets were stacked (up to four layers) to form a ~300-400 μm thick tissue 2 cm in length, organized in a mesh of uniaxial aligned cells. To further demonstrate the effectiveness of this methodology for cultivated meat purposes, we have generated thick and viable constructs using chicken muscle satellite cells (cSCs) and random CAN. This groundbreaking discovery offers a cost-effective and biomimetic solution for cultivating and differentiating muscle cells, forging a crucial link between tissue engineering and the pursuit of sustainable and affordable cultivated meat production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Santos, Guadalupe, Albergaria, Almeida, Moreira, Copola, Paula, Neves, Santos, da Silva, Jorge and Andrade.)

The growing world population, public awareness of animal welfare, environmental impacts and changes in meat consumption leads to the search for novel approaches to food production. Novel foods include products with a new or specifically modified molecular structure, foods made from microorganisms, fungi, algae or insects, as well as from animal cell or tissue cultures. The latter approach is known by various names: "clean meat", "in vitro meat" and "cell-cultured" or "(cell-)cultivated meat". Here, cells isolated from agronomically important species are expanded ex vivo to produce cell biomass used in unstructured meat or to grow and differentiate cells on scaffolds to produce structured meat analogues. Despite the fast-growing field and high financial interest from investors and governments, cultivated meat production still faces challenges ranging from cell source choice, affordable expansion, use of cruelty-free and food-grade media, regulatory issues and consumer acceptance. This overview discusses the above challenges and possible solutions and strategies in the production of cultivated meat. The review integrates multifaceted historical, social, and technological insights of the field, and provides both an engaging comprehensive introduction for general interested and a robust perspective for experts., Competing Interests: Marline Kirsch and Jordi Morales‐Dalmau are employees of Cultimate Foods UG (Berlin/Göttingen). However, the authors declare that they wrote this review objectively and without bias. The company did not influence manuscript preparation. No other conflicts of interest exist., (© 2023 The Authors. Engineering in Life Sciences published by Wiley‐VCH GmbH.)

Cultivated meat is a nascent technology that aims to create an environmentally and animal-friendly alternative to conventional meat. Producing skeletal muscle tissue in an animal-free system allowing for high levels of myofusion and maturation is important for the nutritional and sensorial value of cultivated meat. Alginate is an attractive biomaterial to support muscle formation as it is food-safe, sustainable and cheap and can be crosslinked using non-toxic methods. Although alginate can be functionalized to promote cell attachment, limitations in its mechanical properties, including form, viscosity, and stress relaxation, hinder the cellular capacity for myogenic differentiation and maturation in alginate-based hydrogels. Here, we show that the addition of electrospun short-stranded zein fibers increased hydrogel degradation, resulting in faster compaction, improved cell-gel interaction, and enhanced alignment of bovine muscle precursor cells. We conclude that fiber-hydrogel composites are a promising approach to support optimal formation of 3D constructs, by improving tissue stability and thus prolonging culture duration. Together, this improves muscle-related protein content by facilitating myogenic differentiation and priming muscle organoids for maturation.

Cultured meat, produced by culturing animal cells in vitro, is gaining increasing interest. The first products obtained using this technology were authorized for human consumption in Singapore and the United States, and more are likely to follow in other parts of the world. Therefore, it is important to assess the attitudes toward such meat in various populations and understand the grounds for its acceptance and rejection. The present cross-sectional online study of adult Poles ( n = 1553) aimed to evaluate knowledge of cultured meat, the main reasons and fears associated with its production and consumption, and willingness to buy it and factors influencing such willingness. Most respondents (63%) were familiar with the concept of cultured meat, and 54% declared to purchase it when available. However, concerns over safety were expressed by individuals accepting (39%) and rejecting (49%) such meat. The main motivations for choosing it included limiting animal suffering (76%) and environmental impacts of meat consumption (67%), although over half of responders willing to buy these products were driven by curiosity (58%). Multiple logistic regression revealed that odds (OR; 95%CI) for accepting cultured meat were significantly increased for adults aged 18-40 (1.8; 1.2-2.7); women (1.8; 1.2-2.7); meat eaters (8.7; 5.6-13.6); individuals convinced that animal farming adversely affects the climate (7.6; 3.1-18.3), surface waters (3.1; 1.2-8.1), and air quality (3.0; 1.2-7.6); those familiar with cultured meat concept (4.2, 2.2-8.4); and those revealing high openness to experience (1.7; 1.2-2.4). The results highlight that the Polish population may be moderately ready to accept cultured meat and identify the groups resistant to accepting it. Well-designed and transparent promotion of these products is required to increase the general public's understanding of the potential benefits and challenges of cultured meat technology.

Populations in low- and middle-income countries (LMIC) typically consume less than the recommended daily amount of protein. Alternative protein (AP) sources could help combat malnutrition, but this requires careful consideration of elements needed to further establish AP products in LMIC. Key considerations include technological, nutritional, safety, social, and economic challenges. This perspective analyzes these considerations in achieving dietary diversity in LMIC, using a combination of traditional and novel protein sources with high nutritional value, namely, soy, mycoprotein, and cultivated meat. Technological approaches to modulate the technofunctionality and bitter off-tastes of plant-sourced proteins facilitate processing and ensure consumer acceptance. Economic considerations for inputs, infrastructure for production, and transportation represent key elements to scale up AP. Dietary diversification is indispensable and LMIC cannot rely on plant proteins alone to provide adequate protein intake sustainably. Investments in infrastructure and innovation are urgently needed to offer diverse sources of protein in LMIC., (© 2023 The Authors.)

Agroecology has been proposed as a holistic approach to transform food systems that meet global food requirements with favorable environmental and social impacts. Agroecology relies on science, practices, and social movements that emphasize ecological principles, local knowledge, culture, and traditions to increase the sustainability and equity of the food system. Agroecological practices have demonstrated positive outcomes on food security and nutrition in low- and middle-income countries (LMICs). Agroecology principles can be applied across the food system and could facilitate the integration of certain alternative protein (AP) foods to address multiple issues. In this perspective, agroecological principles were analyzed to compare the suitability of different AP sources: unprocessed/minimally processed legumes, plant-based meats, edible insects, macroalgae (seaweed), fungal biomass, and cultivated meat. Considerations were identified for the feasibility of AP adoption in LMICs within an agroecological framework to provide nutrient-rich and sustainable diets while addressing other principles such as fairness and economic diversity. From this analysis, legumes, simplified plant-based meat analogs such as texturized plant proteins with minimal additives, edible insects, and macroalgae (location dependent) would make excellent nutritional contributions alongside animal-sourced food within LMICs within an agroecological framework. In contrast, highly processed plant-based meats, fungal biomass, and cultivated meat do not align well with agroecological principles for large-scale human consumption within LMICs. Furthermore, the production facilities to make these foods require robust capital investment and there may be issues related to who owns the intellectual property of these technologies. The NOVA classification system categorizes food based on the degree of processing. Our assessment suggests that foods with lower NOVA classification of unprocessed and minimally processed best fit the agroecological principles related to nutrition, agroecosystem, and societal demands for sustainable food systems., Competing Interests: The authors report no conflicts of interest., (© 2023 The Authors.)

When considering options for future foods, cell culture approaches are at the fore, however, culture media to support the process has been identified as a significant contributor to the overall global warming potential (GWP) and cost of cultivated meat production. To address this issue, an artificial intelligence-based approach was applied to simultaneously optimize the GWP, cost, and cell growth rate of a reduced-serum culture media formulation for a zebrafish (ZEM2S cell line) cultivated meat production system. Response surface methodology (RSM) was used to design the experiments, with seven components - IGF, FGF, TGF, PDGF, selenium, ascorbic acid, and serum - selected as independent variables, given their influence on culture media performance. Radial basis function (RBF) neural networks and genetic algorithm (GA) were applied for prediction of dependent variables, and optimization of the culture media formulation, respectively. The results indicated that the developed RBF could accurately predict the GWP, cost and growth rate, with a model efficiency of 0.98. Subsequently, the three developed RBF neural networks predictive models were used as the inputs for a multi-objective genetic algorithm, and the optimal quantities of the independent variables were determined using a multi-objective optimization algorithm. The suggested RSM + RBF + GA framework in this study could be applied to sustainably optimize serum-free media development, identifying the combination of media ingredients that balances yield, environmental impact, and cost for various cultivated meat cell lines., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cultivated meat production requires an efficient, robust and highly optimized serum-free cell culture media for the needed upscaling of muscle cell expansion. Existing formulations of serum-free media are complex, expensive and have not been optimized for muscle cells. Thus, we undertook this work to develop a simple and robust serum-free media for the proliferation of bovine satellite cells (SCs) through Design of Experiment (DOE) and Response Surface Methodology (RSM) using precise and high-throughput image-based cytometry. Proliferative attributes were investigated with transcriptomics and long-term performance was validated using multiple live assays. Here we formulated a media based on three highly optimized components; FGF2 (2 ng/mL), fetuin (600 µg/mL) and BSA (75 µg/mL) which together with an insulin-transferrin-selenium (1x) supplement, sustained the proliferation of bovine SCs, porcine SCs and murine C2C12 muscle cells. Remarkably, cells cultured in our media named Tri-basal 2.0+ performed better than cell cultured in 10% FBS, with respect to proliferation. Hence, the optimized Tri-basal 2.0+ enhanced serum-free cell attachment and long-term proliferation, providing an alternative solution to the use of FBS in the production of cultivated meat., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Mesenchymal stem cell-based cultivated meat is a promising solution to the ecological and ethical problems posed by traditional meat production, since it exhibits a protein content and composition that is more comparable to original meat proteins than any other source of cultivated meat products, including plants, bacteria, and fungi. Nonetheless, the nature and laboratory behavior of mesenchymal stem cells pose two significant challenges for large-scale production: genetic drift and adherent growth in culture. Culture conditions used in the laboratory expose the cells to a selective pressure that causes genetic drift, which may give rise to oncogene activation and the loss of "stemness." This is why genetic and functional analysis of the cells during culture is required to determine the maximum number of passages within the laboratory where no significant mutations or loss of function are detected. Moreover, the adherent growth of mesenchymal stem cells can be an obstacle for their large-scale production since volume to surface ratio is limited for high volume containers. Multi-tray systems, roller bottles, and microcarriers have been proposed as potential solutions to scale-up the production of adherent cells required for cultivated meat. The most promising solutions for the safety problems and large-scale obstacles for cultivated meat production are the determination of a limit number of passages based on a genetic analysis and the use of microcarriers from edible materials to maximize the volume to surface proportion and decrease the downstream operations needed for cultivated meat production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Jaime-Rodríguez, Cadena-Hernández, Rosales-Valencia, Padilla-Sánchez and Chavez-Santoscoy.)

A major challenge of engineering larger macroscale tissues in vitro is the limited diffusion of nutrients and oxygen to the interior. For skeletal muscle, this limitation results in millimeter scale outcomes to avoid necrosis. One method to address this constraint may be to vascularize in vitro -grown muscle tissue, to support nutrient (culture media) flow into the interior of the structure. In this exploratory study, we examine culture conditions that enable myogenic development and endothelial cell survival within tissue engineered 3D muscles. Myoblasts (C2C12s), endothelial cells (HUVECs), and endothelial support cells (C3H 10T1/2s) were seeded into Matrigel-fibrin hydrogels and cast into 3D printed frames to form 3D in vitro skeletal muscle tissues. Our preliminary results suggest that the simultaneous optimization of culture media formulation and cell concentrations is necessary for 3D cultured muscles to exhibit robust myosin heavy chain expression and GFP expression from GFP-transfected endothelial cells. The ability to form differentiated 3D muscles containing endothelial cells is a key step toward achieving vascularized 3D muscle tissues, which have potential use as tissue for implantation in a medical setting, as well as for future foods such as cultivated meats.

Cellular agriculture is an emerging research field of agribiotechnology that aims to produce agricultural products using stem cells, without sacrificing animals or cultivating crops. Cultivated meat, as a representative cellular product of cellular agriculture, is being actively researched due to global food insecurity, environmental, and ethical concerns. This review focuses on the application of stem cells, which are the seeds of cellular agriculture, for the production of cultivated meat, with emphasis on deriving and culturing muscle and adipose stem cells for imitating fresh meat. Establishing standards and safety regulations for culturing stem cells is crucial for the market entry of cultured muscle tissue-based biomaterials. Understanding stem cells is a prerequisite for creating reliable cultivated meat and other cellular agricultural biomaterials. The techniques and regulations from the cultivated meat industry could pave the way for new cellular agriculture industries in the future., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Cellular agriculture is an emerging field rooted in engineering meat-mimicking cell-laden structures using tissue engineering practices that have been developed for biomedical applications, including regenerative medicine. Research and industrial efforts are focused on reducing the cost and improving the throughput of cultivated meat (CM) production using these conventional practices. Due to key differences in the goals of muscle tissue engineering for biomedical versus food applications, conventional strategies may not be economically and technologically viable or socially acceptable. In this review, these two fields are critically compared, and the limitations of biomedical tissue engineering practices in achieving the important requirements of food production are discussed. Additionally, the possible solutions and the most promising biomanufacturing strategies for cellular agriculture are highlighted., Competing Interests: Declaration of interests M.S., J.Q., and A.T. are cofounders of InPrint Bio LLC. The remaining authors have nothing to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The cultivated meat industry, also known as cell-based meat, cultured meat, lab-grown meat, or meat alternatives, is a growing field that aims to generate animal tissues ex-vivo in a cost-effective manner that achieves price parity with traditional agricultural products. However, cell culture media costs account for 55%-90% of production costs. To address this issue, efforts are aimed at optimizing media composition. Systems biology-driven approaches have been successfully used to improve the biomass and productivity of multiple bioproduction platforms, like Chinese hamster ovary cells, by accelerating the development of cell line-specific media and reducing research and development and production costs related to cell media and its optimization. In this review, we summarize systems biology modeling approaches, methods for cell culture media and bioprocess optimization, and metabolic studies done in animals of interest to the cultivated meat industry. More importantly, we identify current gaps in knowledge that prevent the identification of metabolic bottlenecks. These include the lack of genome-scale metabolic models for some species (pigs and ducks), a lack of accurate biomass composition studies for different growth conditions, and 13 C-metabolic flux analysis (MFA) studies for many of the species of interest for the cultivated meat industry (only shrimp and duck cells have been subjected to 13 C-MFA). We also highlight the importance of characterizing the metabolic requirements of cells at the organism, breed, and cell line-specific levels, and we outline future steps that this nascent field needs to take to achieve price parity and production efficiency similar to those of other bioproduction platforms. Practical Application: Our article summarizes systems biology techniques for cell culture media design and bioprocess optimization, which may be used to significantly reduce cell-based meat production costs. We also present the results of experimental studies done on some of the species of interest to the cultivated meat industry and highlight why modeling approaches are required for multiple species, cell-types, and cell lines., (© 2023 Institute of Food Technologists®.)

It is evident that over-consumption of meat can contribute to the emission of hazardous greenhouse gases. One viable way to address such climate impact is to make people become more aware of more sustainable diet options, such as cultivated meat. However, it is challenging to instigate change in people's meat-eating habit, and empirical works have been examining the psychological factors that are related to consumers' willingness to consume cultivated meat. Research has suggested that psychological well-being can play a role in the meaning-making of food consumption, with higher well-being individuals showing more recognition of other sociocultural benefits of consuming food beyond just fulfilling their sustenance needs. As existing works have yet to understand the link between well-being and consumption of novel foods, the current research set out to fill this gap by examining the relationship between people's psychological well-being and their willingness to consume cultivated meat via different reasons (mediators) for consuming cultivated meat. We recruited a representative sample of 948 adults in Singapore to complete an online survey. The study offered the first evidence that there is a positive relationship between people's psychological well-being and their willingness to consume cultivated meat. Further, results revealed that their higher willingness can be motivated by the perception that cultivated meat is as healthy and nutritious, as safe as, and has the same sensory quality as real meat, and is beneficial to the society. This investigation adds to the growing literature on consumer acceptance of cultivated meat by showing the novel finding that well-being and receptivity to cultivated meat is positively linked, and such a positive link can be explained by people's better recognition of the prospective benefits offered by this alternative food., Competing Interests: Declaration of interest There is no conflict of interest in the production of this research., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Cultivated meat is a fast-growing research field and an industry with great potential to overcome the limitations of traditional meat production. Cultivated meat utilizes cell culture and tissue engineering technologies to culture a vast number of cells in vitro and grow/assemble them into structures mimicking the muscle tissues of livestock animals. Stem cells with self-renewal and lineage-specific differentiation abilities have been considered one of the key cell sources for cultivated meats. However, the extensive in vitro culturing/expansion of stem cells results in a reduction in their abilities to proliferate and differentiate. Extracellular matrix (ECM) has been used as a culturing substrate to support cell expansion for cell-based therapies in regenerative medicine due to its resemblance to the native microenvironment of cells. In this study, the effect of the ECM on the expansion of bovine umbilical cord stromal cells (BUSC) in vitro was evaluated and characterized. BUSCs with multi-lineage differentiation potentials were isolated from bovine placental tissue. Decellularized ECM prepared from a confluent monolayer of bovine fibroblasts (BF) is free of cellular components but contains major ECM proteins such as fibronectin and type I collagen and ECM-associated growth factors. Expansion of BUSC on ECM for three passages (around three weeks) resulted in about 500-fold amplification, while cells were amplified less than 10-fold when cultured on standard tissue culture plates (TCP). Moreover, the presence of ECM reduced the requirement for serum in the culture medium. Importantly, the cells amplified on ECM retained their differentiation abilities better than cells cultured on TCP. The results of our study support the notion that monolayer cell-derived ECM may be a strategy to expand bovine cells in vitro effectively and efficiently.

A growing global meat demand requires a decrease in the environmental impacts of meat production. Cultured meat (CM) can potentially address multiple challenges facing animal agriculture, including those related to animal welfare and environmental impacts, but existing cost analyses suggest it is hard for CM to match the relatively low costs of conventionally produced meat. This study analyzes literature reports to contextualize CM's protein and calorie use efficiencies, comparing CM to animal meat products' feed conversion ratios, areal productivities, and nitrogen management. Our analyses show that CM has greater protein and energy areal productivities than conventional meat products, and that waste nitrogen from spent media is critical to CM surpassing the nitrogen use efficiency of meat produced in swine and broiler land-applied manure systems. The CM nutrient management costs, arising from wastewater treatment and land application, are estimated to be more expensive than in conventional meat production. Overall, this study demonstrates that nitrogen management will be a key aspect of sustainability in CM production, as it is in conventional meat systems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Myers, Jaros, Andersen and Raman.)

We present a method of producing bulk cell-cultured fat tissue for food applications. Mass transport limitations (nutrients, oxygen, waste diffusion) of macroscale 3D tissue culture are circumvented by initially culturing murine or porcine adipocytes in 2D, after which bulk fat tissue is produced by mechanically harvesting and aggregating the lipid-filled adipocytes into 3D constructs using alginate or transglutaminase binders. The 3D fat tissues were visually similar to fat tissue harvested from animals, with matching textures based on uniaxial compression tests. The mechanical properties of cultured fat tissues were based on binder choice and concentration, and changes in the fatty acid compositions of cellular triacylglyceride and phospholipids were observed after lipid supplementation (soybean oil) during in vitro culture. This approach of aggregating individual adipocytes into a bulk 3D tissue provides a scalable and versatile strategy to produce cultured fat tissue for food-related applications, thereby addressing a key obstacle in cultivated meat production., Competing Interests: JY, MS, NX, BB, HD, CL, SZ, MR, EL, KZ, GL, JP, DK No competing interests declared, (© 2023, Yuen Jr et al.)

Increases in global meat demands cannot be sustainably met with current methods of livestock farming, which has a substantial impact on greenhouse gas emissions, land use, water consumption, and farm animal welfare. Cultivated meat is a rapidly advancing technology that produces meat products by proliferating and differentiating animal stem cells in large bioreactors, avoiding conventional live-animal farming. While many companies are working in this area, there is a lack of existing infrastructure and experience at commercial scale, resulting in many technical bottlenecks such as scale-up of cell culture and media availability and costs. In this study, we evaluate theoretical cultivated beef production facilities with the goal of envisioning an industry with multiple facilities to produce in total 100,000,000 kg of cultured beef per year or ~0.14% of the annual global beef production. Using the computer-aided process design software, SuperPro Designer®, facilities are modeled to create a comprehensive analysis to highlight improvements that can lower the cost of such a production system and allow cultivated meat products to be competitive. Three facility scenarios are presented with different sized production reactors; ~42,000 L stirred tank bioreactor (STR) with a base case cost of goods sold (COGS) of $35/kg, ~211,000 L STR with a COGS of $25/kg, and ~262,000 L airlift reactor (ALR) with a COGS of $17/kg. This study outlines how advances in scaled up bioreactors, alternative bioreactor designs, and decreased media costs are necessary for commercialization of cultured meat products., (© 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

It is important to have sustainable and edible scaffolds to produce cultivated meat. In this research, three-dimensional (3D) porous scaffolds were developed by soy protein amyloid fibrils for cultivated meat applications. Food-safe biological and physical cross-linking methods using microbial transglutaminase and temperature-controlled water vapor annealing technique were employed to crosslink soy protein amyloid fibrils, resulting in the production of 3D scaffolds. The generated 3D scaffolds had pores with sizes ranging from 50 to 250 μm, porosities of 72-83%, and compressive moduli of 3.8-4.2 kPa, depending on the type of soy protein used in the process (β-conglycinin (7S), glycinin (11S) and soy protein isolate (SPI)). When present with pepsin, these scaffolds can degrade within an hour but remain stable in phosphate-buffered saline for at least 30 days. The soy protein amyloid fibril scaffolds enabled C2C12 mouse skeletal myoblasts proliferate and differentiate without adding cell adhesive proteins or other coatings. The results demonstrate the potential of abundant and inexpensive soy protein amyloid fibrils to be utilized as scaffold materials for cultivated meat in the food industry.

Mesenchymal stem cells (MSCs) can be used as a cell source for cultivated meat production due to their adipose differentiation potential, but MSCs lose their stemness and undergo replicative senescence during expansion in vitro. Autophagy is an important mechanism for senescent cells to remove toxic substances. However, the role of autophagy in the replicative senescence of MSCs is controversial. Here, we evaluated the changes in autophagy in porcine MSCs (pMSCs) during long-term culture in vitro and identified a natural phytochemical, ginsenoside Rg2, that could stimulate pMSC proliferation. First, some typical senescence characteristics were observed in aged pMSCs, including decreased EdU-positive cells, increased senescence-associated beta-galactosidase activity, declined stemness-associated marker OCT4 expression, and enhanced P53 expression. Importantly, autophagic flux was impaired in aged pMSCs, suggesting deficient substrate clearance in aged pMSCs. Rg2 was found to promote the proliferation of pMSCs using MTT assay and EdU staining. In addition, Rg2 inhibited D-galactose-induced senescence and oxidative stress in pMSCs. Rg2 increased autophagic activity via the AMPK signaling pathway. Furthermore, long-term culture with Rg2 promoted the proliferation, inhibited the replicative senescence, and maintained the stemness of pMSCs. These results provide a potential strategy for porcine MSC expansion in vitro.

Cultured meat has emerged as a promising alternative to conventional meat due to its potential to be healthier, more humane, and sustainable. The development of serum-free media by Messmer et al. and the adaptation of soy protein as edible scaffolds by Ben-Arye et al. highlight innovations in this nascent field., Competing Interests: Declaration of interests The authors declare no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Electrospinning emerged as a promising technique to produce scaffolds for cultivated meat in function of its simplicity, versatility, cost-effectiveness, and scalability. Cellulose acetate (CA) is a biocompatible and low-cost material that support cell adhesion and proliferation. Here we investigated CA nanofibers, associated or not with a bioactive annatto extract (CA@A), a food-dye, as potential scaffolds for cultivated meat and muscle tissue engineering. The obtained CA nanofibers were evaluated concerning its physicochemical, morphological, mechanical and biological traits. UV-vis spectroscopy and contact angle measurements confirmed the annatto extract incorporation into the CA nanofibers and the surface wettability of both scaffolds, respectively. SEM images revealed that the scaffolds are porous, containing fibers with no specific alignment. Compared with the pure CA nanofibers, CA@A nanofibers showed increased fiber diameter (420 ± 212 nm vs. 284 ± 130 nm). Mechanical properties revealed that the annatto extract induces a reduction of the stiffness of the scaffold. Molecular analyses revealed that while CA scaffold favored C2C12 myoblast differentiation, the annatto-loaded CA scaffold favored a proliferative state of these cells. These results suggest that the combination of cellulose acetate fibers loaded with annatto extract may be an interesting economical alternative for support long-term muscle cells culture with potential application as scaffold for cultivated meat and muscle tissue engineering., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Santos, Cotta, Santos, Camargos, Carmo, Alcântara, Fleck, Copola, Nogueira, Silva, Andrade, Ferreira and Jorge.)

As consumer acceptance and overall demand for the different types of meat are important determinants of the new balance between conventional and alternative meats, our goal was to approach the changes in meat demand, as affected by the increasingly available alternative meats coupled to the challenges of conventional meat including the meat paradox, with emphasis on the Brazilian scenario. Then, some aspects of the demand for alternative meats are presented, with a brief historical background. As the decisions taken in the present shape this unprecedented revolution in the way we produce and choose whether to eat meat and, if so, which one, the details of the transition to alternative meat chains in Brazil are yet to be written. It seems even more difficult to predict which food protein items will be in higher demand in the next decades, as new products will likely present themselves for their quality as food items as well as for their ethical and environmental attributes., Competing Interests: Declaration of Competing Interest We, the authors of the manuscript entitled ‘Demand changes meat as changing meat reshapes demand: the great meat revolution’, declare no conflict of interests, as also stated in the manuscript file., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The context of accelerated climate change, environmental pollution, ecosystems depletion, loss of biodiversity and growing undernutrition has led human societies to a crossroads where food systems require transformation. New agricultural practices are being advocated in order to achieve food security and face environmental challenges. Cultivated meat has recently been considered one of the most desired alternatives by animal rights advocates because it promises to ensure nutrition for all people while dramatically reducing ecological impacts and animal suffering. It is therefore presented as one of the fairest means of food production for the coming decades, according to utilitarian arguments. However, food security, environmental concerns and animal welfarism guided by a short-term utilitarianism could have techno-optimism bias and could result in some forms of oppression such as anthropocentrism. I argue that there are still deep-rooted moral issues in food systems that are not addressed primarily by lab-grown meat, mainly derived from a loss of sovereignty. Food practices developed in high-tech labs with artificial interventionism constrain the ability of living entities (that are used as food) to flourish on their own terms. This paper aims to explore how sovereignty entitlements for humans and nonhumans are often overlooked by advocates of cultivated meat and the moral challenges it may pose. Accordingly, a more than utilitarian approach framed by ecological and republican justice is proposed here to shed light on some pitfalls of food chains based on cellular agriculture., Competing Interests: Conflict of InterestThe author has no conflicts of interest to declare that are relevant to the content of this article., (© The Author(s) 2022.)

Cultured meat (CM) has emerged as a breakthrough technology, which has the potential to minimise future negative impacts associated with the rapidly growing world population; such as serious environmental, sustainability, global public health, and animal welfare concerns. Recently numerous researchers in academia and companies have been putting significant efforts into scientific and translational development in this field. Since various pillars of CM manufacturing hold substantial translational potential, there has been a steady interest in filing patent applications globally in the past decade. For the first time, we have performed a patentometric analysis for cultured meat by highlighting the current patents landscape, determining prolific organisations and geographical locations active in the area, and identifying the overall outlook for the field. In total, > 190 patents published in the English language were collected from 1997 to 2020 for the analysis. Relevant data were extracted from the patents, including technology focus, country of origin, assignees, priority date, cooperative patent classification (CPC) code, to analyse the current patent landscape.

Global pressure from consumers to improve animal welfare, and reduce microbiological risks or the use of antibiotics pose new challenges for the meat industry. Today's livestock production, despite many undertaken measures, is still far from being sustainable. This forced the need to work on alternative protein types that come from plants, insects, fungi, or cell culture processes. Due to some technical and legal barriers, cultivated meat is not present on the European market, however, in 2020 it was approved in Singapore and in 2022 in the USA. While the technology of obtaining cell cultures from animal muscles has been known and successfully practiced for years, the production of a stable piece of meat with appropriate texture, taste, and smell, is still a problem for several scientific groups related to subsequent companies trying to obtain the highest quality product, in line with the expectations of customers. Although the work on optimal cell meat production has been going on for years, it is still in an early stage, mainly due to several limitations that represent milestones for industrial production. The most important are: the culture media (without animal serum), which will provide an environment for optimal muscle development, natural or close to natural (but still safe for the consumer) stable scaffolds for growing cells. Here, we review the actual knowledge about the above-mentioned challenges which make the production of cellular meat not yet developed on an industrial scale.

The aim of cellular agriculture is to use cell-culturing technologies to produce alternatives to agricultural products. Cultured meat is an example of a cellular agriculture product, made by using tissue engineering methods. This study aims to improve the understanding of the potential environmental impacts of cultured meat production by comparing between different bioprocess design scenarios. This was done by carrying out a life cycle assessment (LCA) for a bioprocess system using hollow fiber bioreactors, and utilizing bench-scale experimental data for C2C12 cell proliferation, differentiation and media metabolism. Scenario and sensitivity analyses were used to test the impact of changes in the system design, data sources, and LCA methods on the results to support process design decision making. We compared alternative scenarios to a baseline of C2C12 cells cultured in hollow fiber bioreactors using media consisting of DMEM with serum, for a 16-day proliferation stage and 7-day differentiation stage. The baseline LCA used the average UK electricity mix as the energy source, and heat treatment for wastewater sterilization. The greatest reduction in environmental impacts were achieved with the scenarios using CHO cell metabolism instead of C2C12 cell metabolisim (64-67 % reduction); achieving 128 % cell biomass increase during differentiation instead of no increase (42-56 % reduction); using wind electricity instead of average UK electricity (6-39 % reduction); and adjusting the amino acid use based on experimental data (16-27 % reduction). The use of chemical wastewater treatment instead of heat treatment increased all environmental impacts, except energy demand, by 1-16 %. This study provides valuable insights for the cultured meat field to understand the effects of different process design scenarios on environmental impacts, and therefore provides a framework for deciding where to focus development efforts for improving the environmental performance of the production system., Competing Interests: Declaration of competing interest Hanna Tuomisto and Scott Allan have no conflicts to declare. Marianne Ellis is co-founder of Cellular Agriculture Ltd., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The present paper is a review of the empirical studies on consumer acceptance of novel protein sources in Italy. The authors uncovered the most recent scientific contributions regarding novel plant-based products, and insect-based foodstuffs as well as cultivated meat, in the attempt of framing the main socio-economic drivers and challenges for each product with respect to consumers' perception and acceptance. Novel plant-based foods and edible insects represent the most viable alternatives in the nearest future, as already available (or very close to being) to the Italian consumer and more appealing to vegetarians and vegans. Nevertheless, some intrinsic characteristics of these products have yet to be accepted. The cultivated meat represents an animal source of protein, hence undesirable for some segments of consumers, although described as a good potential substitute for conventional meat. Nevertheless, institutional, technological and cultural barriers are yet to be solved before paving the way for its presence on Italian supermarket shelves., Competing Interests: Declaration of Competing Interest None., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The growth and activity of adherent cells can be enabled or enhanced through attachment to a solid surface. For food and beverage production processes, these solid supports should be food-grade, low-cost, and biocompatible with the cell of interest. Solid supports that are edible can be a part of the final product, thus simplifying downstream operations in the production of fermented beverages and lab grown meat. We provide proof of concept that edible filamentous fungal pellets can function as a solid support by assessing the attachment and growth of two model cell types: yeast, and myoblast cells. The filamentous fungus Aspergillus oryzae was cultured to produce pellets with 0.9 mm diameter. These fugal pellets were inactivated by heat or chemical methods and characterized physicochemically. Chemically inactivated pellets had the lowest dry mass and were the most hydrophobic. Scanning electron microscope images showed that both yeast and myoblast cells naturally adhered to the fungal pellets. Over 48 h of incubation, immobilized yeast increased five-fold on active pellets and six-fold on heat-inactivated pellets. Myoblast cells proliferated best on heat-treated pellets, where viable cell activity increased almost two-fold, whereas on chemically inactivated pellets myoblasts did not increase in the cell mass. These results support the use of filamentous fungi as a novel cell immobilization biomaterial for food technology applications.

The rapid development of technologies for cultured meat production has led to new challenges for producers regarding appropriate communication with future customers in order to deliver products to a viable market. Communication analysis of social media enables the identification of the key characteristics of the monitored topic, as well as the main areas of communication by individual users based on active digital footprints. This study aimed to identify the key characteristics of cultured meat based on communication analysis of the social network Twitter. Communication analysis was performed based on 36,356 Tweets posted by 4128 individual users. This analysis identified the following main communicated characteristics: clean meat, future meat, and sustainable meat. Latent Dittrich allocation identified five communication topics: (1) clean and sustainable products, (2) comparisons with plant-based protein and the impact on agribusiness, (3) positive environmental aspects, (4) cultured meat as an alternative protein, and (5) the regulation of cultured meat.

Biomaterial scaffolds are critical components in cultivated meat production for enabling cell adhesion, proliferation, differentiation and orientation. Currently, there is limited information on the fabrication of edible/biodegradable scaffolds for cultivated meat applications. In the present work, several abundant, naturally derived biomaterials (gelatin, soy, glutenin, zein, cellulose, alginate, konjac, chitosan) were fabricated into films without toxic cross-linking or stabilizing agents. These films were investigated for support of the adhesion, proliferation and differentiation of murine and bovine myoblasts. These biomaterials supported cell viability, and the protein-based films showed better cell adhesion than the polysaccharide-based films. Surface patterns induced cell alignment and guided myoblast differentiation and organization on the glutenin and zein films. The mechanical properties of the protein films were also assessed and suggested that a range of properties can be achieved to meet food-related goals. Overall, based on adherence, proliferation, differentiation, mechanics, and material availability, protein-based films, particularly glutenin and zein, showed the most promise for cultivated meat applications. Ultimately, this work presents a comparison of suitable biomaterials for cultivated meat applications and suggests future efforts to optimize scaffolds for efficacy and cost., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The cultured meat market has been growing at an accelerated space since the first creation of cultured meat burger back in 2013. Substantial efforts have been made to reduce costs by eliminating serum in growth media and improving process efficiency by employing bioreactors. In parallel, efforts are also being made on scaffolding innovations to offer better cells proliferation, differentiation and tissue development. So far, scaffolds used in cultured meat research are predominantly collagen and gelatin, which are animal-derived. To align with cell-based meat vision i.e. environment conservation and animal welfare, plant-derived biomaterials for scaffolding are being intensively explored. This paper reviews and discusses the advantages and disadvantages of scaffold materials and potential scaffolding related to scale-up solution for the production of cultured meat.

Cultured meat (also referred to as cultivated meat or cell-based meat)-CM-is fabricated through the process of cellular agriculture (CA), which entails application of bioengineering, i.e., tissue engineering (TE) principles to the production of food. The main TE principles include usage of cells, grown in a controlled environment provided by bioreactors and cultivation media supplemented with growth factors and other needed nutrients and signaling molecules, and seeded onto the immobilization elements-microcarriers and scaffolds that provide the adhesion surfaces necessary for anchor-dependent cells and offer 3D organization for multiple cell types. Theoretically, many solutions from regenerative medicine and biomedical engineering can be applied in CM-TE, i.e., CA. However, in practice, there are a number of specificities regarding fabrication of a CM product that needs to fulfill not only the majority of functional criteria of muscle and fat TE, but also has to possess the sensory and nutritional qualities of a traditional food component, i.e., the meat it aims to replace. This is the reason that bioengineering aimed at CM production needs to be regarded as a specific scientific discipline of a multidisciplinary nature, integrating principles from biomedical engineering as well as from food manufacturing, design and development, i.e., food engineering. An important requirement is also the need to use as little as possible of animal-derived components in the whole CM bioprocess. In this review, we aim to present the current knowledge on different bioengineering aspects, pertinent to different current scientific disciplines but all relevant for CM engineering, relevant for muscle TE, including different cell sources, bioreactor types, media requirements, bioprocess monitoring and kinetics and their modifications for use in CA, all in view of their potential for efficient CM bioprocess scale-up. We believe such a review will offer a good overview of different bioengineering strategies for CM production and will be useful to a range of interested stakeholders, from students just entering the CA field to experienced researchers looking for the latest innovations in the field.