High throughput and effective transfection of cells using innovative methods and materials

Cell transfection is one of the most powerful techniques within cell therapy, regenerative medicine, cell biology and the life sciences. The most effective gene transfer protocols require viral vectors or electroporation, which have major drawbacks such as safety concerns and low-throughput, respectively, limiting their range of applicability as well as clinical impact. Replacing the viral systems used in CAR-T cell therapies for patients is a major goal in the field that would transform the emerging potential of cell-based therapies. Optimising transfection protocols for hard-to-transfect cell types will impact research and life sciences fields, opening a milieu of new applications and possibilities involving cell-based products.
Within this Enterprise Doctoral Training Partnership MRC iCASE PhD studentship, a PhD student will use a combination of state-of-the-art material technologies to design and optimise transfection assay protocols for cells such as cardiac cells and immune cells. The studentship will be under the supervision of Prof Molly Stevens, Professor of Biomedical Materials at Imperial College London, within the Department of Materials, Department of Bioengineering and Institute for Biomedical Engineering. Stevens has been recognised by more than 25 major international awards, including the 2016 Clemson Award from the Society for Biomaterials and the 2014 European Group of the Year from the Life Science Awards. The Stevens Group performs multidisciplinary research on bio-inspired materials for applications in regenerative medicine and biosensing. This studentship will be co-supervised by Prof Sian Harding, Professor of Cardiac Pharmacology and Director of Imperial's British Heart Foundation Centre of Excellence. This studentship is in partnership with TTP plc, a SME in the UK that develops new technologies and research instrumentation.
Increasing evidence is proving that mechanical deformation and stress on cells can impact and stimulate transfection. Recently, Stevens published a new nanotechnological platform called "nanoneedles", which could deliver sensitive bio-cargoes such as nucleic acids intramuscularly. These "nanoneedles" can deliver or extract intracellular cargoes within seconds without inducing detrimental perturbations. This research was published in Nature Materials (Front Cover feature), Advanced Materials and ACS Nano. Recently, Stevens published a study in Advanced Materials on the biomechanical forces the "nanoneedles" induce in cells when delivering cargoes, specifically by concurrently activating of caveolae- and clathrin-mediated endocytosis, alongside micropinocytosis, using electron and scanning ion conductance microscopy and molecular biology techniques. The ease of use, elevated biocompatibility and efficient delivery capabilities of "nanoneedles" enable them as viable material platforms to induce transfection, when compared to electroporation. However, here we aim to maximise and support the "nanoneedles" as a high throughput system. For this, Stevens is currently collaborating with TTP plc, who have designed a piezoelectric system, comprising micron-sized arrays of holes, originally designed for aerosolising droplets, but more recently demonstrated for bioprinting and cell manipulation (subject of patent application EP2997125 Method and Device for deformation and/or fragmentation). We have already obtained preliminary data showing improving transfection capabilities when using the piezoelectric system on human mesenchymal stem cells (Figure 1). Within this PhD studentship, we aim to combine these two technologies towards establishing optimal transfection strategies on several cell types, including T-cells as well as cardiac cells, provided by Prof Sian Harding within the National Heart & Lung Institute. This PhD studentship aims to create major impacts in the life sciences, materials science as well as outputting translatable prototypes with improved transfection capabilities.