Project Objectives

• Develop neural progenitor cells engineered to be immune‑compatible and equipped with a dual bioluminescent/fluorescent reporter, enabling non‑invasive, longitudinal tracking after transplantation in mice.
• Establish a deep‑learning‑based behavioural analysis pipeline that can sensitively detect motor recovery after stroke, without the need for painful physical markers or extended testing sessions.
• Integrate electrophysiology, transcriptomics, and neuronal morphology techniques to allow detailed characterisation of graft function and host–graft interactions within the same animal.
• Benchmark the new toolkit by comparing advanced cell therapy, conventional approaches, and placebo conditions to evaluate improvements in precision, reproducibility, and overall scientific value.

3Rs Impact

• Enables repeated, non‑invasive monitoring of transplanted cells, reducing the need for multiple cohorts and lowering the total number of mice required per study.
• Allows multiple levels of analysis — molecular, anatomical, and functional — to be collected from the same animal.
• Replaces invasive and stressful procedures such as immunosuppression, serial surgeries, and joint tattooing.
• Provides more sensitive and reproducible behavioural assessments, improving data quality and decreasing the number of animals needed to detect meaningful effects.
• Overall, this approach could reduce required animal numbers by more than 80% compared with traditional stroke cell-therapy studies.

BACKGROUND

Cell-based therapies are an emerging approach for repairing neurological damage, including deficits caused by stroke. However, current preclinical testing relies heavily on animal models that are invasive, highly variable, and require large numbers of mice. Monitoring transplanted cells often depends on repeated surgical procedures, immunosuppression, or end-point histology at multiple time points, increasing both variability and animal distress. Traditional behavioural tests also have limited sensitivity and reproducibility, making it difficult to accurately assess functional recovery.

Recent technological developments provide opportunities to improve the evaluation of cell therapies. Advances in stem-cell engineering now allow the creation of immune-compatible neural progenitor cells that can survive in mice without the need for immunosuppressive drugs. These cells can also be equipped with bioluminescent reporters, enabling their survival and distribution to be monitored non-invasively through the intact skull. In parallel, deep-learning-based behavioural tools offer more detailed assessment of motor recovery that reduces handling stress. By combining these innovations, this project establishes an integrated, more humane, and scientifically robust system for testing regenerative cell therapies in vivo.