Scaling up the production of cancer organoids for use in drug discovery

类器官在药物开发方面提供了一条潜在的途径；允许减少动物试验，提供更接近人类解剖功能的模型，并提供更复杂的测试机会bbb。在类器官生长中，需要一个允许3D培养的框架。当前的培养程序使用矩阵形式的支持矩阵。Matrigel是由小鼠肉瘤细胞产生的。由于成本高，并且由于小鼠肉瘤不容易在一致的庄园中获得，因此在工业规模上使用这种方法具有挑战性。由于矩阵是一种自然产物，它是内在可变性的来源。该项目将专注于开发和测试一种完全或部分替代的合成基质，以取代或与Matrigel结合使用，为类器官的生长提供更一致的环境。所述合成替代生物材料在类器官生产效率方面至少需要与Matrigel相匹配。使用合成替代品在一致性方面带来好处，减少了批次之间的可变性。合成矩阵可以允许更简单的存储和处理条件，以允许增加应用和使用。这反过来又会增加可调性，因为更多的条件将是可行的。随着合成基质的成功使用，类器官在成熟过程中可以提供更好的结构和力学性能。单独的矩阵不能形成自支撑结构，添加一个合成矩阵可以提供这种能力。自支撑结构也将增加类器官生产的可扩展性，提供适合生物反应器[5]的生长基础。预期的结果是开发一种将被测试和优化的新型水凝胶；从而在工业规模上生产的类器官的一致性和质量都有所提高的基质。在研究中，将研究优化类器官生长的其他途径，包括：测试乳酸对类器官生长的影响；并测试缺氧条件的影响。这些将与合成基质的优化一起，以增加在工业规模上使用类器官的便利性。这些发现有望最大限度地促进类器官的生长。这项研究的好处在于测试新的潜在药物，即治疗不同组织内的癌症。这种检测方式为个性化医疗提供了机会；更高质量的测试；降低开发成本；减少临床试验中的问题；减少动物试验提供了伦理利益，并通过使用更接近现实的人体组织提高了结果的可靠性。通过专注于药物发现的医疗保健优化，这些潜在的好处与EPSRC优先领域保持一致，同时提供更高的可负担性，从而实现更具包容性的医疗保健。a . Lancaster和J. a . Knoblich，“培养皿中的器官发生：使用类器官技术模拟发育和疾病”，《科学》，第345卷，第345期。2014年7月，doi: 10.1126/science.1247125.[2]。Fatehullah， S. H. Tan和N. Barker，“类器官作为人类发育和疾病的体外模型”，Nat. Cell Biol。，第18卷，no。3、艺术。否。3 . 2016年3月，doi: 10.1038/ nccb3312。[3]“生物材料和组织工程- EPSRC网站”。https://epsrc.ukri.org/research/ourportfolio/researchareas/biomaterials/（2020年9月30日访问）。C. Kibbey，“EHS肉瘤的维持和基质制备”，J. Tissue Cult。方法，第16卷，第6期。3, pp. 227-230, sept . 1994, doi: 10.1007/BF01540656.[5]N。Gjorevski等人，“肠道干细胞和类器官培养的设计基质”，《自然》，第539卷，第539期。7630年,艺术。否。7630,2016年11月，doi: 10.1038/nature20168.[6]K。Daniel，《EPSRC医疗技术战略总结》，第12页

Organoids pose a potential route in terms of drug development; allowing a reduction in animal testing, provision of a model more closely resembling human anatomical function, and presenting more complex testing opportunities[1].In organoid growth a framework to allow 3D culture is required. Current culture procedure uses a supporting matrix in the form of Matrigel. Matrigel is produced from mouse sarcoma cells. The use of this at an industrial scale is challenging due to high cost, and low availability as mouse sarcoma is not easily sourced in a consistent manor.As Matrigel is a natural product it is a source of inherent variability. This project will focus on the development and testing of a full or partial replacement synthetic matrix to be used in place of or in combination with Matrigel to provide a more consistent environment for organoid growth. Said synthetic alternative biomaterial will be required to at least match Matrigel alone in efficiency of organoid production[2], [3].Use of a synthetic alternative brings benefits in terms of consistency with reduced batch to batch variability. A synthetic matrix could allow more simplistic storage and handling conditions to allow increased application, and usage. In turn could increase tunability as more conditions would be viable[4].With the successful use of synthetic matrix, increased structural and mechanical properties could be provided to the organoids during maturation. Matrigel alone cannot form a self-supporting structure, the addition of a synthetic matrix could provide this ability. A self-supporting structure would also increase the scalability of organoid production providing a growth foundation which would be suitable within a bioreactor[5].The intended result is development of a novel hydrogel which will be tested and optimised; resulting in a matrix providing an increase in both consistency, and quality of organoids produced on an industrial scale. Within the research other avenues of optimisation of organoid growth will be investigated including: testing the effect of lactate on organoid growth; and testing the effect of hypoxic conditions. These will sit alongside the optimisation of a synthetic matrix to increase the ease of organoid use on an industrial scale.These finding will hope to maximise organoid growth. Benefits of this research lie in testing of new potential drugs namely for the treatment of cancers within differing tissues. Testing this way provides opportunities in personalisation of medicine; higher quality testing; reduction in development costs; decreased issues in clinical trials; and reduction in animal testing which provides ethical benefits as well as increasing the reliability of results through the use of human tissue which provides closer representation to actuality. These potential benefits align with the EPSRC priority areas through healthcare optimisation focusing on drug discovery, alongside the intention to provide increased affordability resulting in more inclusive healthcare[6].[1]M. A. Lancaster and J. A. Knoblich, 'Organogenesis in a dish: Modeling development and disease using organoid technologies', Science, vol. 345, no. 6194, Jul. 2014, doi: 10.1126/science.1247125.[2]A. Fatehullah, S. H. Tan, and N. Barker, 'Organoids as an in vitro model of human development and disease', Nat. Cell Biol., vol. 18, no. 3, Art. no. 3, Mar. 2016, doi: 10.1038/ncb3312.[3]'Biomaterials and tissue engineering - EPSRC website'. https://epsrc.ukri.org/research/ourportfolio/researchareas/biomaterials/ (accessed Sep. 30, 2020).[4]M. C. Kibbey, 'Maintenance of the EHS sarcoma and Matrigel preparation', J. Tissue Cult. Methods, vol. 16, no. 3, pp. 227-230, Sep. 1994, doi: 10.1007/BF01540656.[5]N. Gjorevski et al., 'Designer matrices for intestinal stem cell and organoid culture', Nature, vol. 539, no. 7630, Art. no. 7630, Nov. 2016, doi: 10.1038/nature20168.[6]K. Daniel, 'EPSRC Healthcare Technologies Strategy Summary', p.12