A Novel Single-Cell Culture Approach for Tracking Phenotypic Heterogeneity in vSMCs

Dr. Mairi Sandison (Biomedical Engineering) and Dr. Michele Zagnoni (Electronic & Electrical Engineering) at the University of Strathclyde have developed a novel single-cell culture approach for tracking phenotypic heterogeneity in vascular smooth muscle cells (vSMCs), changes that underlie cardiovascular diseases. They and doctoral student Ellis Smith (funded by Medical Research Scotland) developed a microwell array for the long-term confinement of the progeny of single vSMCs, exploiting Lipidure®-CM as a highly effective cell repellent coating. They also exploited CELLBANKER® cryopreservation media to preserve freshly isolated vSMCs, increasing the number of experiments that can be performed from a single tissue.

• Lipidure® treatment of microwell walls effectively confined strongly migratory cells to their own microwell for at least 3 weeks, even under highly confluent conditions when alternative treatments failed. It remained effective even after freezing cell-loaded microwell arrays.
• This enabled reliable tracking of phenotypic changes in large numbers of single cells and their progeny over time.
• CELLBANKER® cryopreservation media significantly improved survival rates of freshly isolated myocytes. Whilst the post-thaw viability of primary cells was significantly reduced by cryopreservation in DMSO-based media (78% viability rate), and to a lesser extent in Cellbanker 2 (85%), high levels of viability were maintained in Cellbanker 1 (95%).
• CELLBANKER® 1&2 significantly outperformed DMSO-based protocols when assessing the growth of frozen primary cells. Following 1 week of culture, vastly reduced cell growth was obtained with DMSO-based media (5% of that of freshly isolated cells), with clear improvements when using Cellbanker® 1&2 (20% & 27% ).

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