In Vitro Meat

体外肉
  • 文章类型: Journal Article
    养殖肉正在成为一种新型食品,可以以可持续的方式提供动物蛋白质。许多先前的研究采用各种类型的支架来开发具有与屠宰肉相似特性的养殖肉。然而,没有讨论重要的特性,如风味,即使它们决定了食物的质量。风味特征根据在烹饪时通过美拉德反应产生挥发性化合物的氨基酸和糖的量和类型而显著变化。在这项研究中,开发了一种风味可切换的支架,仅在烹饪温度模仿屠宰肉类的美拉德反应时释放肉类风味化合物。通过将可转换风味化合物(SFC)引入明胶基水凝胶中,我们制造了一种功能性支架,可以增强养殖肉的芳香特性。温度响应性SFC在细胞培养期间稳定地保持在支架中,并且可以在烹饪温度下释放。令人惊讶的是,用这种风味可切换的支架制造的养殖肉表现出与牛肉相似的风味模式。这项研究提出了一种策略,通过开发一种功能性支架来开发具有增强的感官特征的培养肉,该支架可以模仿传统肉的天然烹饪风味。
    Cultured meat is emerging as a new type of food that can provide animal protein in a sustainable way. Many previous studies employed various types of scaffolds to develop cultured meat with similar properties to slaughtered meat. However, important properties such as flavor were not discussed, even though they determine the quality of food. Flavor characteristics vary dramatically depending on the amount and types of amino acids and sugars that produce volatile compounds through the Maillard reaction upon cooking. In this study, a flavor-switchable scaffold is developed to release meaty flavor compounds only upon cooking temperature mimicking the Maillard reaction of slaughtered meat. By introducing a switchable flavor compound (SFC) into a gelatin-based hydrogel, we fabricate a functional scaffold that can enhance the aromatic properties of cultured meat. The temperature-responsive SFC stably remains in the scaffold during the cell culture period and can be released at the cooking temperature. Surprisingly, cultured meat fabricated with this flavor-switchable scaffold exhibits a flavor pattern similar to that of beef. This research suggests a strategy to develop cultured meat with enhanced sensorial characteristics by developing a functional scaffold which can mimic the natural cooking flavors of conventional meat.
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  • 文章类型: Journal Article
    培养肉的成本有效开发的关键因素是可在无血清条件下培养的细胞系,以降低生产成本。培养肉中的血红素补充剂模仿原始肉的风味和颜色。这项研究引入了从棒状杆菌产生的细菌提取物,该细菌提取物通过定向进化被选择用于高血红素表达。正常的猪细胞系,PK15用于施用细菌血红素提取物作为补充剂。与先前的研究一致,我们观察到PK15对血红素提取物在10mM或更高的细胞毒性。然而,长期暴露后,PK15适于耐受高达40mM的血红素。对这些血红素适应的PK15细胞(PK15H)的RNA-seq分析揭示了一组改变的基因,主要参与细胞增殖,新陈代谢,和炎症。我们发现细胞色素P450家族1亚家族A,多肽1(CYP1A1),乳过氧化物酶(LPO),谷胱甘肽过氧化物酶5(GPX5)在PK15H血红素剂量依赖性上调。当我们将血清从2%连续降低到无血清时,我们得出了用5-10mM血红素提取物暂时维持的PK15H亚群。总之,我们的研究报告了一种可在高血红素培养基中培养的猪细胞,该细胞可在无血清条件下维持,并提出了在这一适应过程中起关键作用的标记基因.
    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.
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  • 文章类型: Journal Article
    作为细胞培养鱼的关键成分,基于鱼皮明胶(FSG)的细胞支架为细胞生长提供支撑结构,扩散,和差异化。然而,基于FSG的支架存在潜在的致敏性风险。在这项研究中,通过相分离法制备3D可食用支架,其接触角小于90°,这表明支架有利于细胞的粘附。此外,溶胀率大于200%,暗示着支持细胞生长的巨大潜力。序列同源性分析表明FSG易于与胶原类似物发生交叉反应。此外,建立了食物过敏模型,表明用鳕鱼FSG灌胃的小鼠表现出更高水平的特异性抗体,肥大细胞脱颗粒,血管通透性,和肠屏障损伤比那些用pangasius和tilapiasFSG灌胃的肠屏障损伤。其较高的变应原性可能归因于较高数量的耐消化线性表位。此外,较高的水解程度与线性表位的暴露有关,以促进与IgE的结合,这也是维持鳕鱼FSG较高的致敏性的原因。这项研究阐明了细胞培养鱼的致敏性风险,进一步的研究将集中在基于FSG的细胞支架的致敏性降低上。
    As a key component of cell-cultured fish, fish skin gelatin (FSG)-based cell scaffold provides support structures for cell growth, proliferation, and differentiation. However, there are potential allergenicity risks contained in FSG-based scaffolds. In this study, 3D edible scaffolds were prepared by phase separation method and showed a contact angle of less than 90°, which indicated that the scaffolds were favorable for cell adhesion. Besides, the swelling ratio was greater than 200%, implying a great potential to support cell growth. The sequence homology analysis indicated that FSG was prone to cross-reaction with collagen analogues. Additionally, a food allergic model was constructed and represented that mice gavaged with cod FSG exhibited higher levels of specific antibodies, mast cell degranulation, vascular permeability, and intestinal barrier impairment than those gavaged with pangasius and tilapias FSG. Its higher allergenicity might be attributed to a higher number of digestion-resistant linear epitopes. Moreover, the higher hydrolysis degree linked to the exposure of linear epitopes to promote the combination with IgE, which was also responsible for maintaining the higher allergenicity of cod FSG. This study clarifies allergenic risks in cell-cultured fish and further study will focus on the allergenicity reduction of FSG-based cell scaffolds.
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  • 文章类型: Journal Article
    在养殖肉(CM)生产中,支架通过帮助细胞粘附发挥重要作用,增长,分化,和对齐。结缔组织和肌肉组织中纤维微结构的存在引起了组织工程领域的极大兴趣,并引发了研究人员实施支架技术的兴趣。支架技术正在进行广泛的研究工作,以实现生物医学研究的真正肉类结构,并取代无血清CM的生产。由动物衍生的生物材料制成的支架在复制细胞外基质(ECM)方面是有效的,因此,应该把重点放在利用动物副产品上。正确识别和利用植物来源的支架生物材料可能有助于增加除动物来源外的多种选择,并通过支架降低CM生产成本。此外,像静电纺丝这样的技术,改进的静电纺丝和3D生物打印应专注于创建3D多孔支架,以模拟肌肉组织的ECM并形成真正的肉样结构。这篇综述讨论了与结构化CM生产相关的尖端支架技术和可食用生物材料的最新进展。
    In cultured meat (CM) production, Scaffolding plays an important role by aiding cell adhesion, growth, differentiation, and alignment. The existence of fibrous microstructure in connective and muscle tissues has attracted considerable interest in the realm of tissue engineering and triggered the interest of researchers to implement scaffolding techniques. A wide array of research efforts is ongoing in scaffolding technologies for achieving the real meat structure on the principality of biomedical research and to replace serum free CM production. Scaffolds made of animal-derived biomaterials are found efficient in replicating the extracellular matrix (ECM), thus focus should be paid to utilize animal byproducts for this purpose. Proper identification and utilization of plant-derived scaffolding biomaterial could be helpful to add diversified options in addition to animal derived sources and reduce in cost of CM production through scaffolds. Furthermore, techniques like electrospinning, modified electrospinning and 3D bioprinting should be focused on to create 3D porous scaffolds to mimic the ECM of the muscle tissue and form real meat-like structures. This review discusses recent advances in cutting edge scaffolding techniques and edible biomaterials related to structured CM production.
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  • 文章类型: Journal Article
    养殖肉,这包括在实验室里种植肉,而不是饲养动物,在可持续性方面提供了潜在的好处,健康,和动物福利相比,传统的肉类生产。然而,养殖肉的生产过程包括几个阶段,每个都有潜在危险,需要仔细监测和控制。从原始动物和周围环境收集的初始细胞中存在微生物污染风险。在细胞增殖过程中,危害可能包括来自培养基成分的化学残留物,如抗生素和生长因子,以及生物反应器灭菌不当导致的微生物问题。在分化阶段,细胞变成肌肉组织,潜在危害包括脚手架材料的残留物,微载体,和媒体组件。最终成熟和收获阶段可能会受到非无菌条件的环境污染,设备,或工人处理,如果不保持适当的无菌条件。这篇综述探讨了在培养肉类生产过程中必须监测和控制的关键微生物和化学危害。它描述了一些常规和新兴的新技术,可用于检测养殖肉类中的微生物和化学危害。该审查还概述了当前围绕养殖肉类不断发展的监管格局,并解释了如何通过先进的分析技术对微生物和化学危害进行彻底的检测和表征可以提供关键数据,以帮助开发强大的,专门为养殖肉类行业量身定制的循证食品安全法规。建议实施新的数字食品安全方法,以进一步研究灵敏有效地检测养殖肉类中的微生物和化学危害。
    Cultured meat, which involves growing meat in a laboratory rather than breeding animals, offers potential benefits in terms of sustainability, health, and animal welfare compared to conventional meat production. However, the cultured meat production process involves several stages, each with potential hazards requiring careful monitoring and control. Microbial contamination risks exist in the initial cell collection from source animals and the surrounding environment. During cell proliferation, hazards may include chemical residues from media components such as antibiotics and growth factors, as well as microbial issues from improper bioreactor sterilization. In the differentiation stage where cells become muscle tissue, potential hazards include residues from scaffolding materials, microcarriers, and media components. Final maturation and harvesting stages risk environmental contamination from nonsterile conditions, equipment, or worker handling if proper aseptic conditions are not maintained. This review examines the key microbiological and chemical hazards that must be monitored and controlled during the manufacturing process for cultured meats. It describes some conventional and emerging novel techniques that could be applied for the detection of microbial and chemical hazards in cultured meat. The review also outlines the current evolving regulatory landscape around cultured meat and explains how thorough detection and characterization of microbiological and chemical hazards through advanced analytical techniques can provide crucial data to help develop robust, evidence-based food safety regulations specifically tailored for the cultured meat industry. Implementing new digital food safety methods is recommended for further research on the sensitive and effective detection of microbiological and chemical hazards in cultured meat.
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  • 文章类型: Journal Article
    如今,由于全球变暖的同时加剧,食品工业正面临挑战,人口,和食物消费。作为合成生物学与食品科学的结合,为了解决这些问题,新型合成食品得到了高度的关注。然而,这些新颖的食物可能会引起与人类健康有关的潜在风险。四种新型合成食品,包括植物性食品,养殖肉,发酵食品,和以微藻为基础的食物,在研究中进行了综述。原始食物来源,消费者接受度,这些食物的优点和缺点进行了讨论。此外,潜在风险因素,比如营养,生物,和化学危险因素,对与这些食物相关的食物进行了描述和分析。此外,当前的检测方法(例如,酶联免疫吸附测定,生物传感器,色谱,聚合酶链反应,等温扩增,和微流体技术)和处理技术(例如,微波处理,欧姆加热,蒸汽爆炸,高静水压力,超声,冷等离子体,和超临界二氧化碳)进行了严格的审查和讨论。尽管如此,继续创新和开发新的检测和加工技术,以有效评估这些新型合成食品并确保其安全至关重要。最后,简要介绍了提高这些食品质量的方法。它将为食品工业中新型合成食品的开发和管理提供见解。
    Nowadays, the food industry is facing challenges due to the simultaneous rise in global warming, population, and food consumption. As the integration of synthetic biology and food science, novel synthetic foods have obtained high attention to address these issues. However, these novel foods may cause potential risks related to human health. Four types of novel synthetic foods, including plant-based foods, cultured meat, fermented foods, and microalgae-based foods, were reviewed in the study. The original food sources, consumer acceptance, advantages and disadvantages of these foods were discussed. Furthermore, potential risk factors, such as nutritional, biological, and chemical risk factors, associated with these foods were described and analyzed. Additionally, the current detection methods (e.g., enzyme-linked immunosorbent assay, biosensors, chromatography, polymerase chain reaction, isothermal amplification, and microfluidic technology) and processing technologies (e.g., microwave treatment, ohmic heating, steam explosion, high hydrostatic pressure, ultrasound, cold plasma, and supercritical carbon dioxide) were reviewed and discussed critically. Nonetheless, it is crucial to continue innovating and developing new detection and processing technologies to effectively evaluate these novel synthetic foods and ensure their safety. Finally, approaches to enhance the quality of these foods were briefly presented. It will provide insights into the development and management of novel synthetic foods for food industry.
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  • 文章类型: Journal Article
    种子细胞的大量增殖和肉类结构的模仿对于细胞培养的肉类生产仍然具有挑战性。具有优异的生物相容性,高含水量和孔隙率,水凝胶是生物技术应用中经常研究的锚定依赖性细胞支架材料。在这里,一种基于明胶/藻酸盐/ε-聚-1-赖氨酸(GAL)水凝胶的支架被开发用于骨骼肌细胞,在细胞培养肉生产中具有很大的应用前景。在这项工作中,水凝胶GAL-4:1,由明胶(5%,w/v),海藻酸盐(5%,w/v)和ε-聚-1-赖氨酸(摩尔比与藻酸盐:4:1)根据11.29±1.94kPa的杨氏模量选择作为细胞支架,令人满意的剪切稀化性能和合适的多孔组织结构。市售的C2C12小鼠骨骼肌成肌细胞和猪肌肉干细胞(PMuSCs),在3D打印的支架中培养。细胞表现出很强的附着能力,诱导后的增殖和分化,显示出高生物相容性。此外,用GAL-4:1水凝胶和新鲜提取的PSuSC进行细胞生物打印。提取的PMuSCs在3D支架上表现出高活力并显示早期成肌(结蛋白),表明GAL水凝胶作为3D细胞构建支架的巨大潜力。总的来说,我们开发了一种新型的GAL水凝胶作为培养肉研究的3D打印生物活性平台。
    The mass proliferation of seed cells and imitation of meat structures remain challenging for cell-cultured meat production. With excellent biocompatibility, high water content and porosity, hydrogels are frequently-studied materials for anchorage-dependent cell scaffolds in biotechnology applications. Herein, a scaffold based on gelatin/alginate/ε-Poly-l-lysine (GAL) hydrogel is developed for skeletal muscle cells, which has a great prospect in cell-cultured meat production. In this work, the hydrogel GAL-4:1, composed of gelatin (5 %, w/v), alginate (5 %, w/v) and ε-Poly-l-lysine (molar ratio vs. alginate: 4:1) is selected as cell scaffold based on Young\'s modulus of 11.29 ± 1.94 kPa, satisfactory shear-thinning property and suitable porous organized structure. The commercially available C2C12 mouse skeletal myoblasts and porcine muscle stem cells (PMuSCs), are cultured in the 3D-printed scaffold. The cells show strong ability of attachment, proliferation and differentiation after induction, showing high biocompatibility. Furthermore, the cellular bioprinting is performed with GAL-4:1 hydrogel and freshly extracted PMuSCs. The extracted PMuSCs exhibit high viability and display early myogenesis (desmin) on the 3D scaffold, suggesting the great potential of GAL hydrogel as 3D cellular constructs scaffolds. Overall, we develop a novel GAL hydrogel as a 3D-printed bioactive platform for cultured meat research.
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  • 文章类型: Journal Article
    一种新的协议可以通过控制肌肉细胞之间沉积的脂肪水平来定制实验室培养的肉的味道。
    A new protocol can customize the flavor of lab-grown meat by controlling the level of fat deposited between muscle cells.
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  • 文章类型: Journal Article
    栽培肉的出现为传统动物农业提供了可持续和道德的替代方案,强调其在食品工业中的重要性。生物材料支架是培养肉生产中的关键组件,可实现细胞粘附。扩散,分化,和取向。虽然对支架生物材料有广泛的研究,将它们应用于种植肉类生产会带来明显的挑战,每种材料都有自己的优点和缺点。这篇综述总结了最近五年用于细胞培养肉的支架生物材料,详细说明它们各自的优缺点。我们建议未来的研究方向,并为支持可扩展的支架提供建议,成本效益高,和安全的优质肉类生产。此外,我们强调种植肉类面临的商业挑战,包括生物反应器设计,细胞培养基,以及监管和食品安全问题。总之,这篇综述为栽培肉类生产领域的研究人员和公司提供了全面的指导和有价值的见解。
    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.
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  • 文章类型: Journal Article
    目前关于养殖肉的研究主要集中在离体肌肉组织重建,但是缺乏肌肉内脂肪的形成,这是决定口味的关键因素,纹理,和营养成分。因此,将脂肪掺入养殖肉类具有优越的价值。在这项研究中,我们利用鸡成纤维细胞在3D中的成肌/脂肪转分化来产生肌肉质量并将脂肪沉积到相同的细胞中,而无需共培养或混合不同的细胞或脂肪物质。将永生化的鸡胚胎成纤维细胞植入到水凝胶支架中,并在3D中进行细胞增殖和成肌转分化以产生全切肉模拟物。与2D相比,在3D基质中生长的细胞显示出升高的肌生成和胶原蛋白产生。我们进一步诱导了转分化肌细胞中的脂肪沉积,并且可以控制甘油三酸酯的含量以匹配并超过鸡肉的水平。基因表达分析表明,谱系特异性和多功能信号都可能有助于肌肉/脂肪基质的产生。总的来说,我们能够精确地调节肌肉,脂肪,和细胞外基质含量根据平衡或专门的肉类偏好。这些发现为定制的培养肉生产提供了新的途径,具有所需的肌内脂肪含量,可以定制以满足消费者的不同需求。
    Current studies on cultured meat mainly focus on the muscle tissue reconstruction in vitro, but lack the formation of intramuscular fat, which is a crucial factor in determining taste, texture, and nutritional contents. Therefore, incorporating fat into cultured meat is of superior value. In this study, we employed the myogenic/lipogenic transdifferentiation of chicken fibroblasts in 3D to produce muscle mass and deposit fat into the same cells without the co-culture or mixture of different cells or fat substances. The immortalized chicken embryonic fibroblasts were implanted into the hydrogel scaffold, and the cell proliferation and myogenic transdifferentiation were conducted in 3D to produce the whole-cut meat mimics. Compared to 2D, cells grown in 3D matrix showed elevated myogenesis and collagen production. We further induced fat deposition in the transdifferentiated muscle cells and the triglyceride content could be manipulated to match and exceed the levels of chicken meat. The gene expression analysis indicated that both lineage-specific and multifunctional signalings could contribute to the generation of muscle/fat matrix. Overall, we were able to precisely modulate muscle, fat, and extracellular matrix contents according to balanced or specialized meat preferences. These findings provide new avenues for customized cultured meat production with desired intramuscular fat contents that can be tailored to meet the diverse demands of consumers.
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