Engineering extracellular vesicles derived from human mesenchymal stem cells

工程化源自人间充质干细胞的细胞外囊泡

• 批准号: 2113032
• 金额: --
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2113032
• 关键词: Engineering; extracellular; vesicles; derived; human

Engineering extracellular vesicles derived from human mesenchymal stemcellsExtracellular vesicles (EVs) are small, membranous, secreted carriers of proteinsand nucleic acids that enable cell-cell communication. They are evolutionarilyconserved (from yeast to human) and there is a growing interest in the use ofEVs in diverse biotechnological applications, particularly those secreted byintrinsically regenerative cell types such as adult stem cells. We haveengineered several immortalised human clonal mesenchymal stem cell (MSC)lines and demonstrated that they produce different types of EVs. In theorytherefore, these immortalised human cell lines may be used to produce aninexhaustible supply of MSC-EVs. However, being able to control the efficiencyof EV generation, the EV cargo and target cell type would have distinctbiotechnological advantages. This project will build on the mechanisticknowledge in yeast genetics and EV biology to develop bioengineered MSC-EVproduction platforms.Specific objectives are to:-Characterise native MSC-EVs and optimise their isolation.-Determine the underlying mechanisms to help direct and/or enrich cargo-loading in MSC-EVs-Engineer MSC-EVs to target specific cell types.The project will be co-supervised by Professor Paul Genever and Dr. ChrisMacDonald who have broad expertise in MSC biology, EV production, stem celland yeast model systems, endosomal trafficking and genetic engineering. Keytechniques will include cell culture, lentiviral transduction and over-expressionanalyses, transcriptomics, proteomics, bioinformatics and in vitro assays of EVfunction.

工程化源自人间充质干细胞的细胞外囊泡细胞外囊泡 (EV) 是一种小型膜状分泌性蛋白质和核酸载体，可实现细胞间通讯。它们在进化上是保守的（从酵母到人类），并且人们对在各种生物技术应用中使用电动汽车越来越感兴趣，特别是那些由内在再生细胞类型（例如成体干细胞）分泌的电动汽车。我们设计了几种永生化人类克隆间充质干细胞 (MSC) 系，并证明它们能够产生不同类型的 EV。因此，从理论上讲，这些永生化的人类细胞系可用于生产取之不尽的 MSC-EV。然而，如果能够控制电动汽车的发电效率，电动汽车货物和目标细胞类型将具有独特的生物技术优势。该项目将建立在酵母遗传学和 EV 生物学的机械知识基础上，开发生物工程 MSC-EV 生产平台。具体目标是：-表征天然 MSC-EV 并优化其分离。-确定帮助指导和/或丰富 MSC-EV 中的货物装载的基本机制-设计 MSC-EV 以针对特定细胞类型。该项目将是 由 Paul Genever 教授和 ChrisMacDonald 博士共同指导，他们在 MSC 生物学、EV 生产、干细胞和酵母模型系统、内体运输和基因工程方面拥有广泛的专业知识。关键技术将包括细胞培养、慢病毒转导和过表达分析、转录组学、蛋白质组学、生物信息学和 EV 功能体外测定。