3D differentiation conditions for the production of cultured meat from livestock stem cells

全球对肉类的需求正在稳步增长（3%/年），人口增长进一步加剧了这一趋势，预计到2050年将达到100亿。畜牧业肉类生产使用了全球约70%的耕地，导致温室气体（牛肉）和大量用水的增加。根据全球需求的预期增长，到世纪中期，我们将没有足够的地球资源为世界人口提供肉类。因此，迫切需要生产营养、美味和可持续肉类的替代方法。细胞农业部门最近成为实验室肉类生产的可能解决方案，但用于生产培养肉类的生物工程解决方案仍处于起步阶段。在这个项目中，我们将利用我们在干细胞生物学（RA），生物材料/基质生物学（CM）和合成化学（NRT）方面的互补专业知识，开发一种技术创新的方法来生产培养肉。肉是一种复杂的组织，肌肉细胞是主要的细胞成分，并补充了较小比例的脂肪和结缔组织。开发用于生产在体外重现骨骼肌组织的肌细胞的方法是创造替代肉类来源的基础，然后可以进一步工程化以包含其他细胞类型并提供营养和美味的产品。我们将测试这一假设，即细胞外环境的调制可以有一个积极的影响肌肉和脂肪细胞的命运specification.During胚胎发育，组织规格是细胞和细胞外基质（ECM）之间的复杂的相互作用的结果包围他们。该基质由以不同形式存在的蛋白质和聚糖组成，其确切组合赋予每个组织在机械和生物化学性质方面的独特特征。细胞对这些信号做出反应，因为它们形成了专门的组织，如肌肉和脂肪，通常会重新建模基质。在体外，干细胞分化传统上利用动物来源的基质制剂Matrigel。然而，该产品具有显著的局限性，包括缺乏再现性、材料的难处理的复杂性以及在尝试创建无动物成分的产品时的3R限制。使用我们小组先前优化的用于培养小鼠和人类多能干细胞的肽水凝胶，我们可以选择性地引入基质成分，以测试这些成分对动物干细胞分化为肌肉和脂肪的影响。重要的是，可以独立地进行基质蛋白或聚糖的添加以改变水凝胶的机械性质，从而允许单独评估这两个关键因素的影响。

Global demand for meat is steadily increasing (3%/year), and is further exacerbated by population growth, which is estimated to reach 10 billion by 2050. Livestock meat production uses ~70% of global arable land, contributes to increases in greenhouse gases (beef) and considerable water use. Based on the anticipated increase in global demand we will have insufficient planetary resources to provide meat to the world population by the middle of the century. Thus, alternative methods for the production of nutritious, palatable and sustainable meat are urgently needed. The cellular agriculture sector has recently emerged as a possible solution to the production of meat in the laboratory, however bioengineering solutions for the production of cultured meat are still in their infancy. In this project we will take advantage of our complementary expertise in stem cell biology (RA), biomaterials/matrix biology (CM) and synthetic chemistry (NRT) to develop a technologically innovative approach to the production of cultured meat. Meat is a complex tissue, with muscle cells representing the main cellular component, and complemented with smaller proportions of adipose and connective tissue. Developing methods for producing muscle cells that recapitulate the organisation of skeletal muscle in vitro is the foundation for the creation of an alternative source of meat that could then be further engineered to contain other cell types and deliver a nutritious and palatable product. We will test the hypothesis that modulation of the extracellular environment can have a positive effect on muscle and fat cell fate specification.During embryonic development, tissue specification is the result of a complex interplay between cells and the extracellular matrix (ECM) that surrounds them. That matrix is composed of proteins and glycans that exist in different forms, the exact combination of which gives each tissue unique characteristics with regard to both mechanical and biochemical properties. Cells respond to these signals as they form specialised tissues such as muscle and fat, often re-modelling the matrix as they do so. In vitro, stem cell differentiation has traditionally made use of the animal derived matrix preparation, Matrigel. However, this product has significant limitations including a lack of reproducibility, the intractable complexity of the material and the 3Rs limitations when trying to create an animal-component free product. Using a peptide hydrogel previously optimised by our group for culture of mouse and human pluripotent stem cells, we can selectively introduce matrix components to test the impact these have on the differentiation of animal stem cells to muscle and fat. Importantly, the addition of matrix proteins or glycans can be carried out independently to altering the mechanical properties of the hydrogel, allowing separate evaluation of the impact of both of these critical factors.