Meat and Medicine: Regenerative programming of animal and human stem cells for engineered skeletal muscle

肉类和医学：动物和人类干细胞的再生编程用于工程骨骼肌

• 批准号: 2434424
• 金额: --
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2434424
• 关键词: Meat; Medicine; Regenerative; programming; animal

Background: Pluripotent (PSC) and multipotent (such as mesenchymal; MSCs) stem cells hold great promise for the treatment of human skeletal muscle trauma and diseases, but also for the generation of new approaches to generate tissue for food consumption. However, it remains challenging to convert PSCs or MSCs to skeletal muscle cells, and the mechanisms by which the master regulatory transcription factors, promotes muscle stem (satellite) cell identity are not yet fully understood. However it is clear that viral or transgenic expression of transcription factors (TFs) such as Pax7 or MyoD can effectively induce the muscle program in these cell types.Approach: We have developed a non-viral method to deliver genes to tissues, cells or stem cells. This will be the focus of the PhD, to deliver TF genes to stem cells in order to mediate there programming into myogenic precursors and to expand and use those cells to generate tissue of regenerative grafts and as meat-substitutes. You will use PSC and MSC lines as proof of concept, employing iPSCs from pig, sheep and cow to generate species specific muscle tissue for food applications. For clinical use you will employ human iPSCs, MSCs and bone marrow and liposuction aspirates to program these tissue stem cells into muscle for with cell-therapies or for tissue engineering and implant applications. The genes, duration of expression, if serial dosages are required and the culture conditions will require optimisation and assessment. The seeding of scaffolds with generated progenitors or directly programming in 3D will be needed to generate tissue which will also be studied. Interestingly we have electrical stimulation set ups that can be used to physiologically testcontraction of engineered muscle and there is potential to move to in vivo testing depending on the project with collaborators at the RCSI (Dublin, Ireland) and JHU (Baltimore, USA).Location: The project will be carried out in the new state-of-the-art Biodiscovery institute at University Park (James Dixon) and the School of Biosciences at Sutton Bonington Campus (Ramiro Alberio). The use of human and animal cells of this application will be transformative and is likely to have significant commercial value and interaction with stakeholders. Presently the MoD (through the DSTL) are funding bone and fat reconstructive technologies using gene delivery in the Dixon group and this PhD could add impact and synergy to these projects reciprocally.Techniques and opportunities: The project will use cell/stem cell culture, QPCR, immunolabelling, flow cytometry, single cell RNAseq, vector design and construction, gene delivery, scaffolds, tissue engineering, electrical stimulation, in vivo translation.References to learn more:1. Lilja et al. Pax7 remodels the chromatin landscape in skeletal muscle stem cells. PLoS ONE 12(4):e0176190.2. Darabi R and Perlingueiro R (2016). Derivation of Skeletal Myogenic Precursors from Human Pluripotent Stem Cells Using Conditional Expression of PAX7. Methods in Mol Biol 1357:423-439

背景：多能（PSC）和多能（例如间充质；MSC）干细胞在治疗人类骨骼肌创伤和疾病方面具有巨大的前景，而且还可以用于产生用于生成食用组织的新方法。然而，将 PSC 或 MSC 转化为骨骼肌细胞仍然具有挑战性，并且主调控转录因子促进肌肉干（卫星）细胞身份的机制尚未完全了解。然而，很明显，Pax7 或 MyoD 等转录因子 (TF) 的病毒或转基因表达可以有效诱导这些细胞类型中的肌肉程序。方法：我们开发了一种非病毒方法，将基因传递到组织、细胞或干细胞。这将是博士的重点，将 TF 基因传递给干细胞，以介导其编程为肌源性前体细胞，并扩展和使用这些细胞来产生再生移植组织并作为肉类替代品。您将使用 PSC 和 MSC 系作为概念验证，利用猪、羊和牛的 iPSC 生成用于食品应用的物种特异性肌肉组织。在临床应用中，您将使用人类 iPSC、MSC 和骨髓以及抽脂抽吸物将这些组织干细胞编程到肌肉中，用于细胞疗法或组织工程和植入应用。基因、表达持续时间、是否需要连续剂量以及培养条件都需要优化和评估。需要用生成的祖细胞接种支架或直接进行 3D 编程来生成也将进行研究的组织。有趣的是，我们有电刺激装置，可用于对工程肌肉的收缩进行生理测试，并且根据与 RCSI（爱尔兰都柏林）和 JHU（美国巴尔的摩）合作者的项目，有可能转向体内测试。地点：该项目将在大学公园（詹姆斯迪克森）和学院的最先进的新生物发现研究所进行 萨顿博宁顿校区（拉米罗阿尔贝里奥）的生物科学。该应用对人类和动物细胞的使用将是变革性的，并且可能具有重大的商业价值以及与利益相关者的互动。目前，国防部（通过 DSTL）正在资助 Dixon 小组使用基因传递的骨骼和脂肪重建技术，该博士可以相互增加这些项目的影响和协同作用。技术和机会：该项目将使用细胞/干细胞培养、QPCR、免疫标记、流式细胞术、单细胞 RNAseq、载体设计和构建、基因传递、支架、组织工程、电学 刺激，体内翻译。了解更多参考文献：1．莉莉娅等人。 Pax7 重塑骨骼肌干细胞中的染色质景观。 《公共科学图书馆 ONE》12(4)：e0176190.2。 Darabi R 和 Perlingueiro R (2016)。使用 PAX7 的条件表达从人多能干细胞中衍生骨骼肌源前体。 Mol Biol 中的方法 1357:423-439