CPNs™ in Regenerative Medicine and Stem Cell Tracking

As part of our ongoing BBSRC-funded iCASE studentship with the University of Liverpool, we are exploring the use of CPNs™ for in vivo live cell imaging in regenerative medicine for stem cell tracking. The goal of the project is to use CPNs™ to better understand cell activity, which is crucial in the development of cell-based therapies.

We will exploit the multimodal imaging capabilities and enhanced stability of CPNs™ for single cell tracking over 14 days to improve understanding of cell activity which may then lead to development of improved cell-based therapies. Thanks to their powerful brightness, magnetic core and availability in a range of colours, CPNs™ could in the future be used in a variety of key imaging techniques. In addition, their stability means they can be stored for long periods with no loss of brightness.

A. Still images from 3D view of a U87 Glioblastoma spheroid with RFP labelled histones (Red nuclei). Cells were seeded with CPN™ 510A (Green) and incubated for 48 hours. 3D View
B. Still images from 3D view of a U87 Glioblastoma spheroid with H2B-RFP labelled nuclei (Red). Cells were seeded with CPN™ 550s (Yellow) and incubated for 24 hours, 48 hours and 72 hours. 

3D View Images taken on the Andor Dragonfly Spinning Disk Confocal microscope at 40x oil objective using the 488 nm (CPNs™) and 541 nm (H2B-RFP) excitation lasers (taken in collaboration with Suzannah Tasker at the University of Liverpool).

By using dual labelling with a contrasting agent, CPNs™ could possibly be used in bioluminescence imaging (BLI), fluorescence imaging (FI) and magnetic resonance imaging (MRI) to track single cells, both in two and three dimensions. The images produced with fluorescence could aid our understanding of how cells interact with host tissue, how they differentiate and their survival and migratory capabilities.

BLI and MRI have been shown to be effective for non-invasive in vivo imaging of engineered tumours and metastasis in the same model. Using CPNs™ in these types of studies has the potential to improve in vivo imaging capability, providing further insight into cell activity, and ultimately informing the development of cell-based therapies.

We have found that CPNs™ can be used to effectively label glioblastoma (U87 cell line) and human embryonic stem cells (H9 cell line), with exciting wider implications for studying regenerative therapies.


 C. 3D video of a U87 Glioblastoma spheroid labelled with RFP nuclei (Red) and CPN™  510s (Green). Cells seeded with CPN™ 510As and incubated for 48 hours. 3D z-stack video taken on the Andor Dragonfly Spinning Disk Confocal microscope at 40x oil objective using the 488 nm excitation laser. D. Undifferentiated H9 hESCs cells labelled with CPN™ 680 (Red). Colonies were seeded and incubated with CPN™ 680s (Red) for 24 hours, 48 hours and 72 hours. Cell were washed with PBS each day and imaged at each timepoint. Images taken on Andor Dragonfly Spinning Disk Confocal microscope at 25x water objective (taken in collaboration with Suzannah Tasker at the University of Liverpool).