Optimisation and scale-up of nanovibrational stimulation to control the differentiation of adult stem cells

纳米振动刺激控制成体干细胞分化的优化和放大

• 批准号: 2442032
• 金额: --
• 依托单位: University of Strathclyde
• 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: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2442032
• 关键词: Optimisation; scale; up; nanovibrational; stimulation

Nano-amplitude vibration (dubbed 'Nanokicking') has previously been shown to stimulate osteogenic differentiation of mesenchymal stem cells (MSCs). Specifically 1 kHz vibration with 30 nm+ vibration is sufficient to drive osteogenesis and mineralisation in vitro. Bone is the second most transplanted tissue (behind blood) and now these cells are currently being developed as a potential therapy for bone repair due to the clinical need for off-the-shelf replacements for bone autograft. The current bioreactor system capable of stimulating 40-50 million cells per batch but supplying future clinical trials will require billions of cells per batch. This project will examine whether nanovibration can be scaled up and whether this method of stimulation can be used to generate other differentiated cell types. Optimising the vibration parameters (frequency, amplitude, force on cells) will enable these questions to be explored and further progress towards new clinical cell therapies to be made. The aim of the project is to determine optimal values for specific MSC differentiation phenotypes and test if these conditions can be delivered at scale to enable technology transfer from the lab to a clinical environment.The existing bioreactor is focused on 1 kHz, 30 nm vibration but it is not well understood how these parameters tie into mechanotransductive signalling and whether other MSC progenitors could be generated by altering the vibration conditions (e.g. chondrocytes and myocytes). During this project a new vibration bioreactor will be developed to accurately test the differentiation potential of MSCs under varying mechanical loading. In vitro analysis of MSCs, and related mechanosensitive cell types, will investigate how frequency relates to induced cellular tension and stiffness whilst examining the upregulation of cellular markers for each MSC lineage. If new phenotypes can be produced then the optimal vibration conditions will be applied and tested in new scalable cell culture systems such as hollow fibre bioreactors and cell stacks to demonstrate potential for therapeutic cell manufacture.

纳米振幅振动（称为“Nanokicking”）先前已被证明可以刺激间充质干细胞（MSC）的成骨分化。具体而言，1 kHz 振动和 30 nm+ 振动足以驱动体外成骨和矿化。骨骼是第二大移植组织（仅次于血液），由于临床需要现成的自体移植骨替代品，因此这些细胞目前正在开发作为骨修复的潜在疗法。目前的生物反应器系统每批能够刺激 40-5000 万个细胞，但供应未来的临床试验将需要每批数十亿个细胞。该项目将研究纳米振动是否可以扩大规模，以及这种刺激方法是否可以用于产生其他分化的细胞类型。优化振动参数（频率、振幅、细胞力）将使这些问题得到探索，并在新的临床细胞疗法方面取得进一步进展。该项目的目的是确定特定 MSC 分化表型的最佳值，并测试这些条件是否可以大规模提供，从而使技术从实验室转移到临床环境。现有的生物反应器专注于 1 kHz、30 nm 振动，但目前尚不清楚这些参数如何与机械传导信号联系在一起，以及是否可以通过改变振动条件（例如，振动条件）产生其他 MSC 祖细胞。 软骨细胞和肌细胞）。在该项目期间，将开发一种新的振动生物反应器，以准确测试 MSC 在不同机械负载下的分化潜力。对 MSC 和相关机械敏感细胞类型的体外分析将研究频率与诱导的细胞张力和硬度的关系，同时检查每个 MSC 谱系的细胞标记物的上调。如果可以产生新的表型，那么将在新的可扩展细胞培养系统（例如中空纤维生物反应器和细胞堆）中应用和测试最佳振动条件，以证明治疗性细胞制造的潜力。