The project will develop 96 well microplates with fixed electrospun nanofibre scaffolds for use in 3D cell-based assays. There is strong demand in the pharmaceutical industry for more predictive efficacy and toxicology in vitro assays to reduce both the number of costly drug failures in clinical trials and the number of animals used in pre-clinical testing. Despite significant interest in the use of 3D cell-based assays, 3D techniques have not yet been adopted into mainstream research programmes because current formats do not meet industry requirements for highly consistent, off-the-shelf, easy to use products that are compatible with standard automated handling and imaging equipment. The Electrospinning Company (TECL) has developed an innovative microplate into which is laser welded a fibre scaffold, and which has been shown to support the growth of a model breast cancer cell line in 3D. This project will improve this prototype to meet pharmaceutical industry specifications. Cell loading efficiency will be increased to over 90% by testing scaffolds of different fibre specifications, by coating with proteins that increase cell-scaffold anchorage and by pre-loading with denser medium. The optical properties of the plate will be optimised for use on confocal high content analysers by incorporating a glass base plate of appropriate refractive index, and we will reengineer the fabrication process to deliver this cost-effectively. A protocol to sterilise coated scaffolds without damaging the proteins will be developed. The results of these investigations will be used to design a 96 well microplate which will be validated with cell-based assays to prove the concept that it can support the growth of cells in 3D with high reproducibility batch to batch and well-to-well and with the benefits of 3D cell culture. TECL will work with UK SMEs Avanticell, Orla Protein Technologies and 4titude to access expertise in cell-based assays, protein coating and design and manufacture.