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

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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.