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A key element for the cost-effective development of cultured meat is a cell line culturable in serum-free conditions to reduce production costs. Heme supplementation in cultured meat mimics the original meat flavor and color. This study introduced a bacterial extract generated from Corynebacterium that was selected for high-heme expression by directed evolution. A normal porcine cell line, PK15, was used to apply the bacterial heme extract as a supplement. Consistent with prior research, we observed the cytotoxicity of PK15 to the heme extract at 10 mM or higher. However, after long-term exposure, PK15 adapted to tolerate up to 40 mM of heme. An RNA-seq analysis of these heme-adapted PK15 cells (PK15H) revealed a set of altered genes, mainly involved in cell proliferation, metabolism, and inflammation. We found that cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1), lactoperoxidase (LPO), and glutathione peroxidase 5 (GPX5) were upregulated in the PK15H heme dose dependently. When we reduced serum serially from 2% to serum free, we derived the PK15H subpopulation that was transiently maintained with 5–10 mM heme extract. Altogether, our study reports a porcine cell culturable in high-heme media that can be maintained in serum-free conditions and proposes a marker gene that plays a critical role in this adaptation process. [ABSTRACT FROM AUTHOR]

This review article aims to provide an up-to-date overview of the main determinants of consumers' acceptance of novel foods (new foods and ingredients) in the EU with emphasis on product's intrinsic properties (sensory characteristics) and individual factors (socio-demographics, perceptive, psychological) by adopting a systematic approach following the PRISMA methodology. Case studies on terrestrial (i.e., insects, cultured meat and other animal origin products, plant-based food including mushrooms, plant-based analogues, pulses, and cereals) and aquatic systems (i.e., algae and jellyfish) are included focusing on age-related and cross-national differences in consumer acceptance of novel foods and ingredients. General trends have emerged that are common to all the novel foods analysed, regardless of their aquatic or terrestrial origin. Aspects such as food neophobia, unfamiliarity, and poor knowledge of the product are important barriers to the consumption of novel foods, while healthiness and environmental sustainability perception are drivers of acceptance. Sensory properties are challenging for more familiar ingredients such as plant-based food (e.g., novel food made by pulses, mushrooms, cereals and pseudocereals). Results are discussed in terms of feasibility of introducing these products in the EU food systems highlighting strategies that can encourage the use of new ingredients or novel foods. [ABSTRACT FROM AUTHOR]

Reducing production costs, known as scaling, is a significant obstacle in the advancement of cultivated meat. The cultivation process hinges on several key components, e.g., cells, media, scaffolds, and bioreactors. This study demonstrates an innovative approach, departing from traditional stainless steel or glass bioreactors, by integrating food-grade plant-based scaffolds and thermoplastic film bioreactors. While thermoplastic films are commonly used for constructing fluidic systems, conventional welding methods are cost-prohibitive and lack rapid prototyping capabilities, thus inflating research and development expenses. The developed laser welding technique facilitates contamination-free and leakproof sealing of polyethylene films, enabling the efficient fabrication of macrofluidic systems with various designs and dimensions. By incorporating food-grade plant-based scaffolds, such as rice seeded with bovine mesenchymal stem cells, into these bioreactors, this study demonstrates sterile cell proliferation on scaffolds within macrofluidic systems. This approach not only reduces bioreactor prototyping and construction costs but also addresses the need for scalable solutions in both research and industrial settings. Integrating single-use bioreactors with minimal shear forces and incorporating macro carriers such as puffed rice may further enhance biomass production in a scaled-out model. The use of food-grade plant-based scaffolds aligns with sustainable practices in tissue engineering and cultured-meat production, emphasizing its suitability for diverse applications. [ABSTRACT FROM AUTHOR]

Natural scaffolds have been the cornerstone of tissue engineering for decades, providing ideal environments for cell growth within extracellular matrices. Previous studies have favored animal-derived materials, including collagen, gelatin, and laminin, owing to their superior effects in promoting cell attachment, proliferation, and differentiation compared to non-animal scaffolds, and used immortalized cell lines. However, for cultured meat production, non-animal-derived scaffolds with edible cells are preferred. Our study represents the first research to describe plant-derived, film-type scaffolds to overcome limitations associated with previously reported thick, gel-type scaffolds completely devoid of animal-derived materials. This approach has been employed to address the difficulties of fostering bovine muscle cell survival, migration, and differentiation in three-dimensional co-cultures. Primary bovine myoblasts from Bos Taurus Coreanae were harvested and seeded on alginate (Algi) or corn-derived alginate (AlgiC) scaffolds. Scaffold functionalities, including biocompatibility and the promotion of cell proliferation and differentiation, were evaluated using cell viability assays, immunofluorescence staining, and reverse transcription-quantitative polymerase chain reaction. Our results reveal a statistically significant 71.7% decrease in production time using film-type scaffolds relative to that for gel-type scaffolds, which can be maintained for up to 7 days. Film-type scaffolds enhanced initial cell attachment owing to their flatness and thinness relative to gel-type scaffolds. Algi and AlgiC film-type scaffolds both demonstrated low cytotoxicity over seven days of cell culture. Our findings indicated that PAX7 expression increased 16.5-fold in alginate scaffolds and 22.8-fold in AlgiC from day 1 to day 3. Moreover, at the differentiation stage on day 7, MHC expression was elevated 41.8-fold (Algi) and 32.7-fold (AlgiC), providing initial confirmation of the differentiation potential of bovine muscle cells. These findings suggest that both Algi and AlgiC film scaffolds are advantageous for cultured meat production. [ABSTRACT FROM AUTHOR]

To investigate the influence of food culture on the acceptability of meat substitutes, in 2023 we conducted a questionnaire survey on men and women aged 18–69 years (N = 1681) in Japan, a country with a food culture that regards soya as a meat substitute, focusing on 'meaning' in the theory of practice. Analysis of three groups—a flexible group who accept foods processed from soya as meat, a strict group who do not accept such foods as meat, and an intermediate group—showed that the flexible group tended to be more positive about the consumption of meat substitutes than the other groups. However, the flexible group was not simply more accepting, as they expressed aversion and anxiety in response to other questions. Food culture, i.e., what is understood to be meat, influences the acceptability of alternative meats. When examining the social acceptability of meat alternatives in the future, it is also important to look at social practices and historical and cultural backgrounds in a multilayered way, in addition to factors that have been emphasized in previous studies, such as consumers' decision-making process. [ABSTRACT FROM AUTHOR]

The selection of an appropriate scaffold is imperative for the successful development of alternative animal protein in the form of cultured meat or lab-grown meat. Decellularized tissues have been suggested as a potential scaffold for cultured meat production owing to their capacity to support an optimal environment and niche conducive to cell proliferation and growth. This approach facilitates the systematic development of 3D tissues in the laboratory. Decellularized scaffold biomaterials have characteristics of high biocompatibility, biodegradation, and various bioactivities, which could potentially address the limitations associated with synthetic bio-scaffold materials. The present study involved the derivation and characterization of a decellularized scaffold from mushroom tissue following subsequent assessment of the scaffold's capacity to support myogenic differentiation. Mushroom sections were soaked in nuclease and detergent solution for 4 days. Furthermore, decellularization was confirmed by histology and DAPI staining, which showed the removal of cellular components and nuclei. Myoblast cells were seeded onto decellularized tissue, which exhibited excellent cytocompatibility and promoted myogenic growth and differentiation. The study's findings can serve as a foreground for the generation of an edible and natural scaffold for producing a safe and disease-free source of alternative animal protein, potentially reducing the burden on the health sector caused by conventional animal protein production and consumption. [ABSTRACT FROM AUTHOR]

The widespread commercialization of cultured meat, produced from animal stem cells grown in vitro, faces significant challenges related to technical, regulatory, and social acceptability constraints. Despite advancements in knowledge, the acceptance of this innovation remains uncertain. Understanding individuals' decision-making processes and interpretative patterns is crucial, with media framing playing a key role in shaping attitudes toward cultured meat adoption. This research, focusing on Twitter as a social media platform, examines the impact of media framing on consumer attitudes (cognitive, affective, and conative) regarding cultured meat. Qualitative (content analysis) and quantitative (MANOVA) analyses were conducted on 23,020 posts and 38,531 comments, selected based on media framing or containing relevant attitude components. This study reveals that media-framed posts significantly influence consumer attitudes compared to non-media-framed posts. While different types of media framing (ethical, intrinsic, informational, and belief) exhibit varying impacts on attitude components, posts combining ethical, intrinsic, and informational frames have a more substantial effect on cultured meat acceptability. The belief frame, particularly for the behavioral component, is equally influential. Consumer attitudes toward cultured meat are found to be ambivalent, considering the associated benefits and risks. Nevertheless, the affective component of attitude is notably influenced by posts featuring informational and ethical media frames. This study suggests implications for authorities and businesses, emphasizing the importance of differentiated education and marketing strategies. Advertising messages that combine ethical, intrinsic, and informational frames are recommended. Additionally, this study advocates for regulatory measures governing the production, marketing, and consumption of cultured meat to instill consumer confidence in the industry. By highlighting the significance of beliefs in cultured meat consumption behavior, this research points toward potential exploration of cultural and religious influences in future studies. [ABSTRACT FROM AUTHOR]

Cultured meat presents a possible alternative to conventional meat products and may be used to address growing food demands attributable to global population growth. Thus, a comprehensive technological prospection of the scientific literature related to cultured meat produced by 3D bioprinting is of great interest to researchers. The purpose of this article is to review and analyze published studies related to the biofabrication of cultured meat using 3D bioprinting techniques. The growing number of related publications in recent years highlights that cultured meat has gained traction in the scientific community. Furthermore, private companies and startups have contributed to advancements in the biofabrication of cultured meat for consumption, illustrating that cultured meat as a conventional meat substitute is already becoming reality. However, like any scientific advance, 3D bioprinting of cultured meat faces challenges involving regulation, acceptance, the selection of ideal biomaterials and cell lines, the replacement of fetal bovine serum (FBS), and attaining a texture and nutritional value similar to those of conventional meat. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Cultured meat is one of the meat substitutes produced through tissue engineering and other technologies. Large-scale cell culture is the key for cultured meat products to enter the market. Therefore, this study is aimed to explore the effect of long-term passage in vitro on smooth muscle cells (SMCs) and the effect of transforming growth factor-β1 (TGF-β1) on SMCs in the late passage. Multiple passages lead to the decline of the proliferation rate of SMCs in the proliferation stage and the differentiation ability in the differentiation stage. Transcriptome results showed that the ECM pathway and aging-related signaling pathways were significantly up-regulated in the late passage period. TGF-β1 did not promote SMCs of late passage proliferation at the proliferation stage but promoted the gene and protein expression of collagen as the main protein of the extracellular matrix proteins at the differentiation stage. In addition, proteomic analysis revealed that TGF-β1 promoted the expression of cell adhesion molecules which activate the Hippo signaling pathway and the HIF-1 signaling pathway and further promoted the production of collagen-containing extracellular matrix proteins. This could provide ideas for large-scale production of cultured meat products using SMCs. [ABSTRACT FROM AUTHOR]

As with every new technology, safety demonstration is a critical component of bringing products to market and gaining public acceptance for cultured meat and seafood. This manuscript develops research priorities from the findings of a series of interviews and workshops with governmental scientists and regulators from food safety agencies in fifteen jurisdictions globally. The interviews and workshops aimed to identify the key safety questions and priority areas of research. Participants raised questions about which aspects of cultured meat and seafood production are novel, and the implications of the paucity of public information on the topic. Novel parameters and targets may require the development of new analytical methods or adaptation and validation of existing ones, including for a diversity of product types and processes. Participants emphasized that data sharing of these efforts would be valuable, similar to those already developed and used in the food and pharmaceutical fields. Contributions to such databases from the private and public sectors would speed general understanding as well as efforts to make evaluations more efficient. In turn, these resources, combined with transparent risk assessment, will be critical elements of building consumer trust in cultured meat and seafood products. [ABSTRACT FROM AUTHOR]

Growing research and technological development is making the commercial production of cultured meat as a sustainable alternative to livestock-derived meat an increasing reality. However, to competitively position cultured meat on the food market, appropriate marketing and communication tailored to specific demographics is required. We aimed to define the motives that influence the willingness to include cultured meat in consumption based on age, specifically in Generation Z and Generation Y. To achieve this, data from a questionnaire survey that asked about ethical, ecological and health and safety factors around cultured meat was collected from 740 respondents (301 Generation Z and 439 Generation Y) and analyzed using the Mann–Whitney test and structural equation modeling. Generation Z were significantly more likely than Generation Y (p < 0.05) to consider cultured meat healthier than conventional meat because of the possibility of adjusting the composition and nutrient content. Generation Z were also significantly less concerned than Generation Y (p < 0.05) about the consequences that consuming cultured meat might have on human health. In Generation Z, ethical, ecological and health and safety factors significantly influenced their willingness to consume cultured meat (all p < 0.01). In conclusion, we confirmed the influence of ecological and ethical awareness, as well as health and safety, on willingness to include cultured meat in consumption; these areas could be targeted when marketing cultured meat. [ABSTRACT FROM AUTHOR]

Cultured meat is a new type of green, safe, healthy, and sustainable alternative to traditional meat that will potentially alleviate the environmental impact of animal farming and reduce the requirement for animal slaughter. However, the cultured meat structures that have been prepared lack sufficient tissue alignment. To create a product that is similar in texture and taste to traditional animal meat, muscle stem cells must be organized in a way that imitates the natural structure of animal tissue. Recently, various scaffold technologies and biomaterials have been developed to support the three-dimensional (3D) cultivation and organization of muscle stem cells. Hence, we propose an overview of the latest advancements and challenges in creating three-dimensional scaffolds for the biomanufacturing of cultured meat. [ABSTRACT FROM AUTHOR]

New food technologies such as cultured meat, precision fermentation, and plant-based alternatives may one day supplant traditional modes of animal farming. Nonetheless, very little is known about how traditional animal farmers perceive these new products, despite being directly impacted by their advance. The present study explored the views of livestock farmers regarding emerging protein alternatives. We used a comparison group of omnivorous non-farmers as a frame of reference. Forty-five UK-based livestock farmers and fifty-three non-farmers read an informative vignette about emerging food technologies that reviewed their advantages vis-à-vis intensive animal agriculture. Afterwards, participants rated four products (plant-based burgers; plant-based milk alternatives; cultured beef; animal-free dairy milk) in terms of their personal appeal and how much they represented a positive change to the market. Participants furthermore voiced their agreement or disagreement towards 26 statements representing potential facilitators or barriers to product acceptance. Overall, farmers rated the four products less appealing and less beneficial to the industry compared to non-farmers. Positive change ratings tended to be higher than personal appeal ratings for all products. Both groups tended to agree that the alternatives offered advantages, particularly for the environment, resource use, food security, and animal treatment, though agreement rates were lower for farmers. Farmers tended to perceive more barriers to acceptance than non-farmers, with 'threat to farmers' and 'lack of support to local farmers' of paramount concern to both groups. These findings highlight how farmers' attitudes towards alternative proteins are mixed and, ultimately, shaped by the perceived vulnerability of farming communities. [ABSTRACT FROM AUTHOR]

Simple Summary: The present study examined the potential benefits of male specific-pathogen-free (SPF) White Leghorn embryos in cellular agriculture for sustainable and ethical poultry meat production—addressing traditional farming challenges, including disease outbreaks of Salmonella and Avian influenza. We generated myogenic precursor cells (MPCs) from the thigh muscles of embryos from SPF White Leghorns that tested negative for Salmonella. When male and female embryonic MPCs were cultured in vitro, it was confirmed that genetic sex did not affect the expression of factors related to myogenic regulation factors. Therefore, MPCs from male SPF-laying chicken embryos are promising for developing clean animal-cell-derived protein sources via resource recycling. This study examined the potential benefits of male specific-pathogen-free (SPF) White Leghorn embryos in cellular agriculture for sustainable and ethical poultry meat production—addressing traditional farming challenges, including disease outbreaks of Salmonella and Avian influenza. We isolated myogenic precursor cells (MPCs) from the thigh muscles (Musculus femoris) of 12.5-day-old embryos from 10 SPF White Leghorns that tested negative for Salmonella. We randomly selected MPCs from three males and three females, isolated them using a modified pre-plating (pp) method, and compared their in vitro development. After 1 h (pp1) and 2 h (pp2) of incubation, they were transferred to a new dish to remove fast-adhering cells and cultured (pp3). Isolated MPCs had a 69% positive reaction to Pax7. During proliferation, no differences were observed in PAX7, MYF5, or MYOD expression between the male and female MPCs. However, after five days of differentiation, the expression of late myogenic factors—MYOG and MYF6—significantly increased in all MPCs. Notably, MYOG expression was 1.9 times higher in female than in male MPCs. This impacted MYMK's expression pattern. Despite this, the myotube fusion index did not differ between the sexes. Muscle cells from male SPF-laying chicken embryos are promising for developing clean animal-cell-derived protein sources via resource recycling. [ABSTRACT FROM AUTHOR]

In recent years, a great deal of research has been conducted on consumers' attitudes and beliefs in favor and against alternative proteins (AP). By contrast, a much more limited research effort has been devoted to understanding stakeholders' point of view. The current work provides a first systematic review of the existing literature on stakeholders' beliefs towards alternative protein sources. Moreover, a secondary content analysis was carried out on the selected studies, providing an overview of the major themes emerging from the existing literature in relation to utilitarian, normative, and control beliefs that stakeholders hold towards AP. Results showed that stakeholders' beliefs are very different from those that emerged from previous research on consumers' views. Overall, stakeholders appear much more aware, compared to consumers, of the implications of using alternative proteins in relation to the main pillars of sustainability (economic, environmental, social). Stakeholders' beliefs were grouped into 13 categories, such as "economics", "consumers", and "rules". With respect to future scenarios, they attribute an important role to political institutions, with respect to both economic and communication aspects, which they consider crucial to overcome persistent consumer skepticism. [ABSTRACT FROM AUTHOR]

Cell-cultured meat, which is artificial meat made by in vitro cultivation of animal-derived cells, has attracted a lot of interest as a potential source of protein in the future. Porous hydrogels are crucial components that can be used as an artificial extracellular matrix (ECM) to provide cell growth for generating cultured meat. In this study, we highlight the effects of grape seed extract (proanthocyanidins, PC) on the physicochemical and biological functions (bovine satellite muscle cell (BSC) growth and adhesion) of an edible gelatin (GL)-based hydrogel. The freeze-dried hydrogels had good compressive characteristics with pore sizes ranging from 100 to 300 μm. BSCs were able to grow and attach to porous GL-PC hydrogels. These studies suggested that the developed hydrogels using edible materials and made by employing a low-cost method may serve in the cell growth of muscle cells for cultured meat applications. [ABSTRACT FROM AUTHOR]

Cultured meat is an emerging technology for manufacturing meat through cell culture rather than animal rearing. Under most existing culture systems, the content and maturity of in vitro generated myotubes are insufficient, limiting the application and public acceptance of cultured meat. Here we demonstrated that a natural compound, naringenin (NAR), promoted myogenic differentiation of porcine satellite cells (PSCs) in vitro and increased the content and maturity of generated myotubes, especially for PSCs that had undergone extensive expansion. Mechanistically, NAR upregulated the IGF-1/AKT/mTOR anabolic pathway during the myogenesis of PSCs by activating the estrogen receptor β. Moreover, PSCs were mixed with hydrogels and cultured in a mold with parallel micro-channels to manufacture cultured pork samples. More mature myosin was detected, and obvious sarcomere was observed when the differentiation medium was supplemented with NAR. Taken together, these findings suggested that NAR induced the differentiation of PSCs and generation of mature myotubes through upregulation of the IGF-1 signaling, contributing to the development of efficient and innovative cultured meat production systems. [ABSTRACT FROM AUTHOR]

Meat culturing technology goes beyond laboratory research and materialises in the market. Nonetheless, this technology has raised concerns among Muslim consumers worldwide due to its medium, especially foetal bovine serum (FBS), which originates from blood. Thus, the aim of this research was to determine the halal status of cultured meat by detecting species-specific DNA of bovine serum as one of the media used during meat production. Polymerase chain reaction (PCR) analysis was conducted by targeting mitochondrial cytochrome oxidase II (COII) gene sequences, producing a 165 bp amplicon. The sequences of the primers used were Bovine-F, 5′-CAT CAT AGC AAT TGC CAT AGT CC-3′ and Bovine-R, 5′-GTA CTA GTA GTA TTA GAG CTA GAA TTA G-3′. DNA extraction was conducted using a QIAGEN Blood and Tissue™ commercial kit. The presence study also included a literature review on the Istihalah (transformation) concept in order to determine the halal status of cultured meat. The results revealed that bovine DNA was detected in all samples tested using PCR analysis. Therefore, Istihalah tammah (perfect transformation) does not occur due to the ability of PCR analysis to detect bovine DNA in FBS and is prohibited according to Shariah law. [ABSTRACT FROM AUTHOR]

Simple Summary: Existing food production systems and dietary behaviours are not healthy nor sustainable due to the higher environmental footprint of animal-derived foods. Alternative dietary choices could provide better outcomes. We investigated Generation Z (Gen Z)'s alternative food preferences within a planetary health framework which combines human and ecological well-being. The paper is based on data from a 2021 cross-national survey conducted in the main Australian cities, which explored Gen Z's attitudes towards meat consumption in relation to climate change and alternatives to animal-based proteins. Climate change is seen as a result of human activities by 86% of the survey participants. More than a third (38%) of them believe that livestock production and the consumption of animal-sourced foods are contributing significantly to climate change and environmental deterioration. The remaining majority (62%), however, has a low awareness and understanding of the food systems' impacts, disbelieving that diet is a major contributor to climate change. We discuss how these findings will shape Australia's Gen Z, climate change and the future of alternative proteins. Scientific evidence shows that current food systems are impacting the planet in ways that are unsustainable and detrimental to human health. Various technological advances have been made in response, one of them being the development of new food products known as novel alternative proteins, including cultured meat, plant-based meat analogues, algae- and insect-based foods. The future of these alternative proteins to a large extent depends on consumer acceptance from young people. This study investigates the attitudes of Australia's adult Generation Z (Gen Z), born between 1995 and 2003, regarding climate change and more sustainable food choices. Gen Z is a diverse, important and trendsetting group known for organising globally on causes related to climate, social justice and health. The study of Australia's Gen Z is based on a 2021 cross-national survey in the main Australian cities. It shows that, although 86% of the participants perceive climate change as anthropogenic, only 38% believe that livestock-based foods are contributing significantly to global warming and environmental deterioration. The paper discusses the implications for Gen Z and novel alternative proteins given that the majority of Australia's young people has low awareness of the environmental impacts of food systems and dietary choices. [ABSTRACT FROM AUTHOR]

Cultured meat, or the practice of growing meat from cell culture, has been experiencing rapid advances in research and technology as the field of biotechnology attempts to answer the call to fight climate change and feed a growing global population. A major hurdle for cell-based meat products entering the market in the near-future is their price. The complex production facilities required to make such products will require advanced bioreactor systems, resources such as energy and water, and a skilled labor force, among other factors. The use of insect cells in this process is hypothesized to address some of these costs due to the characteristics that make them more resilient in cell culture when compared to traditional livestock-derived cells. To address the potential for cost savings by utilizing insect cells in the cultivation of protein-enriched foods, here we utilized a techno-economic assessment model. Three different insect cell lines were used in the model. The results indicate that insect cell lines offer potential to significantly reduce the cost per kilogram of cell cultivated meat, along with further opportunities to optimize production processes through technological advances and scaling. [ABSTRACT FROM AUTHOR]

Cultivated meat is an innovative product that promises to provide a consumption experience and nutritional value equivalent to conventional meat, with significantly lower negative impacts. The aim of this study is to analyze the factors that influence the purchase intention of cultivated meat. A quantitative study was conducted with 304 Brazilian consumers. The data were analyzed using structural equation modeling. The study proposed a purchase intention model with aspects related to clean production, animal welfare, food safety, naturalness and the moderating effect of food neophobia. The results showed that clean production and food safety had a positive relationship with the intention to purchase cultivated meat. The naturalness of food, on the other hand, had a negative relationship with purchase intent. Furthermore, the higher the level of food neophobia, the less important the influence of clean production became with regard to cultivated meat purchase intentions. [ABSTRACT FROM AUTHOR]

Cultured meat refers to the production of animal tissue by utilizing the same techniques as tissue engineering through cell culture. Various biomaterials have been designed to serve as in vitro supports for cell viability, growth, and migration. In this study, visible light and dual-crosslinked alginate hydrogels were designed to enable control of the physical and mechanical properties needed for the fabrication of cultured meat scaffolds. We hypothesized that a difference in hydrogel stiffness would influence cell behavior, indicating the efficacy of our processing methods to benefit the cultured meat field. Herein, we synthesized and created: (1) methacrylated alginate (AlgMA) to enable covalent crosslinking via visible light exposure, (2) Methacrylated alginate and arginyl-glycyl-aspartic acid RGD conjugates (AlgMA-RGD), using carbodiimide chemistries to provide cell-binding sites on the material, and (3) designer hydrogels incorporating different crosslinking techniques. The material and mechanical properties were evaluated to determine the structural integrity of the hydrogels, and in vitro cell assays were conducted to verify cytocompatibility and cell adhesion. Gelation, swell ratio, and weight loss calculations revealed longer gelation times for the AlgMA scaffolds and similar physical properties for all hydrogel groups. We showed that by adjusting the polymer concentration and the crosslinking methodology, the scaffold's mechanical properties can be controlled and optimized within physiological ranges. Incorporating dual crosslinking significantly increased the compressive moduli of the AlgMA hydrogels, compared to visible-light crosslinking alone. Moreover, the muscle satellite cells responded favorably to the AlgMA scaffolds, with clear differences in cell density when cultured on materials with significantly different mechanical properties. Our results indicate the usefulness of the dual-crosslinking alginate hydrogel system to support in vitro meat growth. [ABSTRACT FROM AUTHOR]

Environmentally intense and negative consequences relateing to conventional meat production systems have induced some actors to suggest alternative meat sources. Diseases carried by animals, human perception of cruelty to animals, and public health concerns about cardiovascular diseases have provided the basis for the development of cultured meat. The current market is influenced by many factors, including regulators, affordability, religion, and media perception. The existing cultured meat market is also regulated by legislatures, affordability, consumer religion, and the media. Consumer perception is distributed across various aspects, including ethical priorities, nutritional profile of the meat consumed, age-based acceptance, gender differentiation, political orientation, land-based attitude, education status, socioeconomic factors, and familiarity factor with the existing product in the market. Inhibiting barriers reported among consumers—including low naturalness, safety, nutritional concerns, trust, neophobia, economic, and ethical approaches—should be employed as marketing tactics directly to address their respective concerns. Tissue culture, starter cells, printing, and 3D printing are some of the methods currently being used for the production of cultured meat. Similarly, many hybrid technologies are also being used to produce meat-like products to increase consumer familiarity along and market presence. Existing research frameworks have improved the previous mindset of consumers with media coverage, educational frameworks, and the textural attributes of cultured meat. Additional benefits of CUME may include being environmentally friendly with less production of greenhouse gases. However, consumer trust, affordability, improving nutritional status, and widescale adoption are just a few of the parameters that need to be addressed to enhance consumer acceptability of these products. The aim of this article was to analyze the current state of cultured meat and the marketing content challenges and strategies used to advance public acceptance of cultured meat. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Many studies have already addressed the perception of cultured meat, but the segmentation of potential consumers has not been adequately investigated. Based on an online survey in Croatia with 411 valid responses, four socio-demographically characterized segments were identified. The key differentiators between the segments relate to consumers' moral and ethical concerns about cultured meat and their perceptions of the impact of cultured meat production on the economy. The multinomial logistic regression used to describe the segments found that the segments differed from the reference segment (Concerned) in terms of support for public research and funding of cultured meat, the willingness to consume it, and religiosity. These results help provide deeper insight into the profiles of potential consumers of cultured meat. [ABSTRACT FROM AUTHOR]

Featured Application: Decellularized plant-based scaffolds can serve as a key component in affordable scaled cultured meat production systems. They are an affordable, natural, and edible cell carrier and their use in suspension cultures would contribute to nutritional properties of a final meat product. Rapidly expanding skeletal muscle satellite cells with cost-effective methods have been presented as a solution for meeting the growing global demand for meat. A common strategy for scaling cell proliferation employs microcarriers, small beads designed to support anchorage-dependent cells in suspension-style bioreactors. No carrier has yet been marketed for the cultivation of lab-grown meat. The objective of this study was to demonstrate a rapid, food safe, decellularization procedure to yield cell-free extracellular matrix scaffolds and evaluate them as cell carriers for lab grown meat. Broccoli florets were soaked in SDS, Tween-20, and bleach for 48 h. The decellularization process was confirmed via histology, which showed an absence of cell nuclei, and DNA quantification (0.0037 ± 0.00961 μg DNA/mg tissue). Decellularized carriers were sorted by cross sectional area (7.07 ± 1.74 mm2, 3.03 ± 1.15 mm2, and 0.49 ± 0.3 mm2) measured for eccentricity (0.73 ± 0.16). Density measurements of decellularized carriers (1.01 ± 0.01 g/cm) were comparable to traditional microcarriers. Primary bovine satellite cells were inoculated into and cultured within a reactor containing decellularized carriers. Cell adhesion was observed and cell death was limited to 2.55 ± 1.09%. These studies suggested that broccoli florets may serve as adequate edible carrier scaffolds for satellite cells. [ABSTRACT FROM AUTHOR]

Global food systems are under significant pressure to provide enough food, particularly protein-rich foods whose demand is on the rise in times of crisis and inflation, as presently existing due to post-COVID-19 pandemic effects and ongoing conflict in Ukraine and resulting in looming food insecurity, according to FAO. Cultivated meat (CM) and cultivated seafood (CS) are protein-rich alternatives for traditional meat and fish that are obtained via cellular agriculture (CA) i.e., tissue engineering for food applications. Stem and progenitor cells are the building blocks and starting point for any CA bioprocess. This review presents CA-relevant vertebrate cell types and procedures needed for their myogenic and adipogenic differentiation since muscle and fat tissue are the primary target tissues for CM/CS production. The review also describes existing challenges, such as a need for immortalized cell lines, or physical and biochemical parameters needed for enhanced meat/fat culture efficiency and ways to address them. [ABSTRACT FROM AUTHOR]

Most beef cattle in the United States start their lives on pasture and finish them in crowded feedlots, releasing hundreds of pounds of the greenhouse gases methane and nitrous oxide, before they are transported to a slaughterhouse, where they are killed and their bodies are sliced into steaks and ground into hamburgers. Until recently, the alternatives to this system were either meat produced in the less sustainable but more humane method of raising cattle solely on pasture and utilizing smaller-scale slaughterhouses or plant-based meat substitutes. The development of the first cultured beef burger in 2013, produced through tissue engineering, raised the possibility of a newer and better alternative. In this article, I use the example of cultured meat to argue that religion and technology are co-constitutive, that they shape and reshape each other, and that the intersection between religion and technology in meat production has had and continues to have a direct impact on animals raised for meat. Kosher meat, industrial or cultured, exemplifies the complexities in the relationship between religion, technology, and animals and will serve as the example throughout this article. [ABSTRACT FROM AUTHOR]

The world's growing population requires an adequate supply of protein to maintain food security, but animal protein production is limited by the finite resources of land, fresh water, and ocean capacity. Several meat substitutes offer protein alternatives that may improve food security in less-developed economies. However, perceptions of difference in the ethical risk associated with consumption of plant-based substitutes (PM) vs. cultured meat (CM) may affect purchases of these products. This study examined differences in ethical risk perception using online survey data gathered in 2020. An ordered logit technique yielded the probabilities of changes in ethical risk perception influenced by demographic attributes, views about the technology, and adequacy of industry regulations. The results show that consumers associated PM with low ethical risk. Educated consumers were more likely to agree that the ethical risks of CM are higher than PM and to regard PM products as safer than CM. Price sensitivity made consumers more likely to agree that the ethical risks related to CM are higher than those related to PM. Ingredient safety concerns increased the ethical risk perception of CM. Consumers perceiving the meat substitute classification to be unclear were more likely to assign a higher ethical risk to CM than PM. The perception of ethical risk associated with CM was greater than that associated with PM if meat substitute industry regulations were inadequate. The results suggest a need to provide verifiable information about each type of meat substitute as well as transparent and understandable standards and rules before these products can improve protein availability and food security. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Cultured meat production is an evolving method of producing animal meat using tissue engineering techniques. Cells, chemical factors, and suitable biomaterials that serve as scaffolds are all essential for the cultivation of muscle tissue. Scaffolding is essential for the development of organized meat products resembling steaks because it provides the mechanical stability needed by cells to attach, differentiate, and mature. In in vivo settings, extracellular matrix (ECM) ensures substrates and scaffolds are provided for cells. The ECM of skeletal muscle (SM) maintains tissue elasticity, creates adhesion points for cells, provides a three-dimensional (3D) environment, and regulates biological processes. Consequently, creating mimics of native ECM is a difficult task. Animal-derived polymers like collagen are often regarded as the gold standard for producing scaffolds with ECM-like properties. Animal-free scaffolds are being investigated as a potential source of stable, chemically defined, low-cost materials for cultured meat production. In this review, we explore the influence of ECM on myogenesis and its role as a scaffold and vital component to improve the efficacy of the culture media used to produce cultured meat. [ABSTRACT FROM AUTHOR]

Cultured meat is one of a number of alternative proteins which can help to reduce the demand for meat from animals in the future. As cultured meat nears commercialization, research on consumers' perceptions of the technology has proliferated. We build on our 2018 systematic review to identify 26 empirical studies on consumer acceptance of cultured meat published in peer-reviewed journals since then. We find support for many of the findings of our previous review, as well as novel insights into the market for cultured meat. We find evidence of a substantial market for cultured meat in many countries, as well as markets and demographics which are particularly open to the concept. Consumers mostly identified animal- and environment-related benefits, but there is plenty of potential to highlight personal benefits such as health and food safety. The safety of cultured meat and its nutritional qualities are intuitively seen as risks by some consumers, although some recognize potential benefits in these areas. Evidence suggests that acceptance can be increased with positive information, as well as frames which invoke more positive associations. We conclude by arguing that cultured meat will form one part of a varied landscape of future protein sources, each appealing to different groups of consumers to achieve an overall reduction in conventional meat consumption. We acknowledge a range of pro-cultured meat messaging strategies, and suggest that framing cultured meat as a solution to existing food safety problems may be an effective approach to increase acceptance. In the long-term, objections based in neophobia and norm violation will decrease, and widespread acceptance will depend in large part on the price and taste. [ABSTRACT FROM AUTHOR]

The selection of an appropriate scaffold is imperative for the successful development of alternative animal protein in the form of cultured meat or lab-grown meat. Decellularized tissues have been suggested as a potential scaffold for cultured meat production owing to their capacity to support an optimal environment and niche conducive to cell proliferation and growth. This approach facilitates the systematic development of 3D tissues in the laboratory. Decellularized scaffold biomaterials have characteristics of high biocompatibility, biodegradation, and various bioactivities, which could potentially address the limitations associated with synthetic bio-scaffold materials. The present study involved the derivation and characterization of a decellularized scaffold from mushroom tissue following subsequent assessment of the scaffold's capacity to support myogenic differentiation. Mushroom sections were soaked in nuclease and detergent solution for 4 days. Furthermore, decellularization was confirmed by histology and DAPI staining, which showed the removal of cellular components and nuclei. Myoblast cells were seeded onto decellularized tissue, which exhibited excellent cytocompatibility and promoted myogenic growth and differentiation. The study's findings can serve as a foreground for the generation of an edible and natural scaffold for producing a safe and disease-free source of alternative animal protein, potentially reducing the burden on the health sector caused by conventional animal protein production and consumption.

The food industry is constantly challenged to find new ideas to satisfy the increasingly specific consumer demand. However, innovative food products do not always become part of consumption habits or create a market. One of the major sources of resistance to novelty lies in the attitude of the consumer, who in many cases may be suspicious or hostile as a result of specific ideologies, overly attached to tradition, or affected by neophobia. This paper analyzes the construct of food neophobia (the "unwillingness to try new foods") in its phenomenology and its actual power to explain hostility to innovation in the agri-food sector. The limits of the concept, which is not always sufficient to shed light on the many reasons that could underlie the rejection of certain foods, will also be discussed. In addition, we review the recent literature on Europeans' attitude toward novel foods and innovation including Genetically modified organisms (GMOs), cultivated meat and insects as food. This literature reveals a number of paradoxes in consumers' behavior, and in the many complex conditions underpinning the success of innovation in food production. These conditions can only be understood by reconstructing the meanings consumers assign to food, and are often embedded in larger social and political frameworks. [ABSTRACT FROM AUTHOR]

This study examines the willingness to consume a cultured meat burger in Germany. Based on the theory of planned behaviour (TPB), we assessed attitudes, perceived behavioural control, and subjective norms via an online questionnaire. Attitudes were operationalized in this research as general attitudes towards cultured meat and specific attitudes towards a cultured meat burger. Furthermore, the TPB was extended with nutritional-psychological variables including food (technology) neophobia, food disgust, sensation seeking, and green consumption values. In total, 58.4% of the participants reported being willing to consume a cultured meat burger. Using a path model, the extended TPB accounted for 77.8% of the variance in willingness to consume a cultured meat burger. All components of the TPB were significant predictors except general attitudes. The influence of general attitudes was completely mediated by specific attitudes. All nutritional-psychological variables influenced general attitudes. Food technology neophobia was the strongest negative, and green consumption values were the strongest positive predictor of general attitudes. Marketing strategies should therefore target the attitudes of consumers by encouraging the natural perception of cultured meat, using a less technological product name, enabling transparency about the production, and creating a dialogue about both the fears and the environmental benefits of the new technology. [ABSTRACT FROM AUTHOR]

The application of chitosan (CS) and whey protein (WP) alone or in combination in 3D/4D printing has been well considered in previous studies. Although several excellent reviews on additive manufacturing discussed the properties and biomedical applications of CS and WP, there is a lack of a systemic review about CS and WP bio-inks for 3D/4D printing applications. Easily modified bio-ink with optimal printability is a key for additive manufacturing. CS, WP, and WP–CS complex hydrogel possess great potential in making bio-ink that can be broadly used for future 3D/4D printing, because CS is a functional polysaccharide with good biodegradability, biocompatibility, non-immunogenicity, and non-carcinogenicity, while CS–WP complex hydrogel has better printability and drug-delivery effectivity than WP hydrogel. The review summarizes the current advances of bio-ink preparation employing CS and/or WP to satisfy the requirements of 3D/4D printing and post-treatment of materials. The applications of CS/WP bio-ink mainly focus on 3D food printing with a few applications in cosmetics. The review also highlights the trends of CS/WP bio-inks as potential candidates in 4D printing. Some promising strategies for developing novel bio-inks based on CS and/or WP are introduced, aiming to provide new insights into the value-added development and commercial CS and WP utilization. [ABSTRACT FROM AUTHOR]

Cultured meat has been presented as an environmentally friendlier option to conventional meat, but due to the limited data, the studies related to its performance are scarce and based on hypothetical production processes. This work provides a short literature review of the published environmental assessments of cultured meat. The main findings of this critical analysis showed that the lack of real data related to cultured meat decreased the level of accuracy of each study. The missing environmental profile of the process itself, including the proliferation and differentiation phases in bioreactors, along with key ingredients such as growth factors and other recombinant proteins, increase the difficulty of achieving reliable conclusions. In order to bridge the highlighted gaps, a complete production system is modelled and analysed from an engineering and life-cycle perspective. Furthermore, an overview of the supply chains of different products used in the process is provided, together with recommendations on how they should be considered in future life-cycle assessments. In essence, this work provides a structured pathway for upcoming consistent environmental assessments in this field, with the objective of setting the basis to understand the potential of cultured meat. [ABSTRACT FROM AUTHOR]

The dietary protein requirements of almost 9.8 billion people need to be fulfilled in a healthy and sustainable manner by 2050. Meat consumption contributes to 35% of the total protein requirement of the Indian population. Meat intake needs to be sustainable and economical without causing food security and production issues. Consumption of meat in India is projected to rise with an increase in consumer incomes. Hence, novel alternative proteins, including cultured meat (CM) and plant-based meat (PBM), are being developed to satisfy the demand for meat-derived proteins in the diet. This involves the creation of novel PBM/CM products with a similar taste and texture as conventional animal meat with tailor-made nutritional attributes. In this article, we provide critical insights into the technical and business aspects of relevance to production and sustainability encountered by the Indian CM industry at a series of stages that can be termed the CM value chain comprising upstream and downstream processes. We shed light on the need for regulatory authorities and a framework. Consumer concerns towards CM products can be alleviated through effective scientific communication strategies, including prior familiarity, narrative building and transparency, and labelling aspects of CM products. [ABSTRACT FROM AUTHOR]

The need to produce immortal, food-relevant cell lines is one of the most pressing challenges of cellular agriculture, the field which seeks to produce meat and other animal products via tissue engineering and synthetic biology. Immortal cell lines have a long and complicated story, from the first recognized immortal human cell lines taken from Henrietta Lacks, to today, where they are used to assay toxicity and produce therapeutics, to the future, where they could be used to create meat without harming an animal. Although work in immortal cell lines began more than 50 years ago, there are few existing cell lines made of species and cell types appropriate for cultured meat. Cells in cultured meat will be eaten by consumers; therefore, cultured meat cell lines will also require unique attributes not selected for in other cell line applications. Specifically, cultured meat cell lines will need to be approved as safe for consumption as food, proliferate and differentiate efficiently at industrial scales, and have desirable taste, texture, and nutrition characteristics for consumers. This paper defines what cell lines are needed, the existing methods to produce new cell lines and their limitations, and the unique considerations of cell lines used in cultured meat. [ABSTRACT FROM AUTHOR]

Muscle stem cells (MuSCs) isolated ex vivo are essential original cells to produce cultured meat. Currently, one of the main obstacles for cultured meat production derives from the limited capacity of large-scale amplification of MuSCs, especially under high-density culture condition. Here, we show that at higher cell densities, proliferation and differentiation capacities of porcine MuSCs are impaired. We investigate the roles of Hippo-YAP signaling, which is important regulators in response to cell contact inhibition. Interestingly, abundant but not functional YAP proteins are accumulated in MuSCs seeded at high density. When treated with lysophosphatidic acid (LPA), the activator of YAP, porcine MuSCs exhibit increased proliferation and elevated differentiation potential compared with control cells. Moreover, constitutively active YAP with deactivated phosphorylation sites, but not intact YAP, promotes cell proliferation and stemness maintenance of MuSCs. Together, we reveal a potential molecular target that enables massive MuSCs expansion for large-scale cultured meat production under high-density condition. [ABSTRACT FROM AUTHOR]

The world's population continues to increase, meaning we require more consistent protein supply to meet demand. Despite the availability of plant-based protein alternatives, animal meat remains a popular, high-quality protein source. Research studies have focused on cultured meat (meat grown in vitro) as a safe and more efficient alternative to traditional meat. Cultured meat is produced by in vitro myogenesis, which involves the processing of muscle satellite and mature muscle cells. Meat culture efficiency is largely determined by the culture conditions, such as the cell type and cell culture medium used and the biomolecular composition. Protein production can be enhanced by providing the optimum biochemical and physical conditions for skeletal muscle cell growth, while myoblasts play important roles in skeletal muscle formation and growth. This review describes the cell types used to produce cultured meat and the biological effects of various myokines and cytokines, such as interleukin-6, leukemia inhibitory factor, interleukin-4, interleukin-15, and interleukin-1β, on skeletal muscle and myogenesis and their potential roles in cultured meat production. [ABSTRACT FROM AUTHOR]

Cultured meat is an emerging alternative food technology which aims to deliver a more ethical, sustainable, and healthy muscle-tissue-derived food item compared to conventional meat. As start-up companies are rapidly forming and accelerating this technology, many aspects of this multi-faceted science have still not been investigated in academia. In this study, we investigated if bovine satellite cells with the ability to proliferate and undergo myogenic differentiation could be isolated after extended tissue storage, for the purpose of increasing the practicality for cultured meat production. Proliferation of bovine satellite cells isolated on the day of arrival or after 2 and 5 days of tissue storage were analyzed by metabolic and DNA-based assays, while their myogenic characteristics were investigated using RT-qPCR and immunofluorescence. Extended tissue storage up to 5 days did not negatively affect proliferation nor the ability to undergo fusion and create myosin heavy chain-positive myotubes. The expression patterns of myogenic and muscle-specific genes were also not affected after tissue storage. In fact, the data indicated a positive trend in terms of myogenic potential after tissue storage, although it was non-significant. These results suggest that the timeframe of which viable myogenic satellite cells can be isolated and used for cultured meat production can be greatly extended by proper tissue storage. [ABSTRACT FROM AUTHOR]