Development of a Multi-Modal StimulationPlatform for the 3D Culture of Wholly EdibleSkeletal Muscle Tissue

该项目属于EPSRC再生医学创新技术研究领域，属于医疗保健技术主题。工业畜牧业约占全球温室气体排放量的14.5%。随着越来越多的国家承诺到2050年实现净零碳目标，覆盖全球总排放量的88%，减少肉类生产的影响对于实现这些目标至关重要。为了满足日益增长的需求，已经采取了大范围砍伐森林的做法。在2022年的前6个月，超过1200平方公里的巴西亚马逊地区被清理，其中36%的农业砍伐森林仅被分配给牧场，这是被清理土地最大的单一利用案例。这对当地的生态系统和生物多样性产生了灾难性的影响，并消除了全球最大的碳汇之一。此外，饲养和屠宰供人食用的动物存在严重的伦理困境，每年有超过700亿只动物被杀害。提出的解决与肉类生产相关的环境和伦理问题的方法之一是发展养殖肉类。培养的肉是从活的动物身上取来的活检组织，利用组织工程技术，这些细胞被培养成一种产品，复制传统的肉类选择。常春藤农场是这项研究的前沿公司，也是该项目的工业合作伙伴。该项目建议为这一应用部署中空纤维膜生物反应器。与3D生物打印替代品相比，这些技术为养殖肉类生产提供了一个可扩展、低成本的平台。此外，搅拌槽式生物反应器用于大量细胞生产，但缺乏其他生物反应器设计的分化潜力，使其不太适合复制肉类的复杂结构和质地。该项目的总体目标是设计一种中空纤维膜生物反应器平台，该平台利用完全可食用的内部结构，只需最少的下游加工即可适用于养殖肉类产品。中空纤维膜生物反应器非常适合这种应用，因为它们便于体内环境的复制。中空纤维起到血管的作用，运输营养和氧气，同时提供一个支架，细胞可以在上面种植。多种细胞刺激模式，如电信号，将被用来促进细胞分化，以获得所需的最终结构和质地。这些技术在养殖肉类生产中的应用是非常新颖的，将与常春藤农场一起从无到有地发展。核心步骤被视为中空纤维膜的不同材料和制造技术的评估。必须将这些参数与一组最能代表真实动物组织性能的参数进行比较。这对养殖肉类也是一个独特的挑战，因为严格的生物医学应用并不要求考虑质地和味道。因此，生物反应器的设计和制造可以开始，考虑到刺激方法，以最大限度地促进差异化。此外，将有一个重要的数学建模方面，使未来的设计迭代得以优化。在合作者开发的质量传输模型的基础上，这些模型将应用于每个生物反应器内的特定条件，并进行实验验证。此外，该项目与牛津人形生物反应器实验室小组密切相关。这不仅带来了体外建模技术领域的专业知识，还与生物医学行业建立了密切的联系，这意味着可以探索新疗法开发的任何潜在应用。

This project falls within the EPSRC Innovative technologies for regenerative medicine research area, within the Healthcare Technologies Theme.ndustrial livestock farming is responsible for approximately 14.5 % of global greenhouse gas emissions.With an increasing number of nations committing to net zero carbon targets by 2050, covering 88 % of totalglobal emissions, reducing the impact of meat production is critical to meeting these aims. For productionto match the ramping demand, widespread deforestation has been deployed. In the first 6 months of 2022over 1200 square kilometers of the Brazilian Amazon were cleared, with 36 % of agricultural deforestationallotted to cattle pastures alone, the largest single utilisation case for the cleared land. This has disastrouseffects on local ecosystems and biodiversity as well as removing one of the largest global carbon sinks.Additionally, there are significant ethical dilemmas attached to the rearing and slaughter of animals forhuman consumption, with over 70 billion animals killed each year.One of the methods proposed to tackle the environmental and ethical issues associated with meat production is the development of cultured meat. Cultured meat is produced by taking a biopsy from a livinganimal and, using tissue engineering techniques, these cells are cultivated to form a product which replicates traditionally available meat options. Ivy Farm are a company at the forefront of this research andare the industrial partner for this project.This project proposes that hollow fibre membrane bioreactors are deployed for this application. Theseoffer a scalable, low-cost platform for cultured meat production when compared to 3D bioprinting alternatives. Additionally, stirred tank bioreactors are used for large volume cell production, but lack the differentiation potential of other bioreactor designs, making them less well suited to replicating the complexstructures and textures of meat.The overarching aim for the project is to design a hollow fibre membrane bioreactor platform which utilisesa wholly edible internal structure and requires minimal downstream processing before it is suitable fora cultured meat product. Hollow fibre membrane bioreactors are ideally suited to this application asthey facilitate the replication of the in-vivo environment. Hollow fibres serve the role of blood vessels for1nutrient and oxygen transport, while providing a scaffold onto which cells can be seeded.Multiple models of cell stimulation, such as electrical signalling, will be utilised to promote cell differentiation to achieve the desired final structure and texture. The application of these techniques to culturedmeat production is highly novel and will be developed from the ground up alongside Ivy Farm.The central steps are seen as the evaluation of different materials and manufacturing techniques for thehollow fibre membranes. These must be compared against a set of parameters that best represent theperformance of genuine animal tissue. This is also a unique challenge to cultured meat, as considerationsof texture and flavour are not required for strictly biomedical applications. Consequently, bioreactor designand manufacture can be commenced, factoring in stimulation methods to best facilitate differentiation.Furthermore, there will be a significant mathematical modelling aspect, enabling future design iterationsto optimised. Building on mass transport models developed by collaborators, these will be applied tothe specific conditions within each bioreactor, and validated experimentally. Additionally, this project isclosely linked to the Oxford humanoid bioreactor lab group. Not only does this bring expertise in the fieldof in-vitro modelling techniques, but also a close link to the biomedical industry, meaning any potentialapplications to novel therapy development can be explored.