Optimizing Smooth Muscle Cell Cryopreservation for Cardiovascular Research

AMSBIO, in collaboration with the University of Strathclyde (UK), announce the release of a poster that describes a comprehensive assessment of the effects of three different cryopreservation reagents (CELLBANKER® 1, CELLBANKER® 2 and commonly used 10% DMSO) on the viability and proliferative capacity of vascular smooth muscle cells (vSMCs)

Central to understanding the vascular remodelling that underlies cardiovascular diseases is the ability to perform reliable single-cell analysis of vSMCs. Large-scale solutions for single cell analysis and the tracking of cell fate using microarray technologies are well established. However, challenges exist when working with freshly isolated vSMCs, as these quickly deteriorate in buffer and there is a limited time before they lose their native phenotype in culture. To reduce animal use and maximise the number of viable cells, establishing a protocol for robust cryopreservation of native vSMCs is critical.

The poster explains how Dr. Mairi Sandison and Dr. Michele Zagnoni at the University of Strathclyde have developed a novel single-cell culture approach for tracking phenotypic heterogeneity in vSMCs, to advance our understanding of the cellular sub-populations that are drivers of disease.  They, and doctoral student Ellis Smith, developed a sophisticated microwell array designed for the long-term confinement of single vSMC progeny. This approach leverages the exceptional cell-repellent properties of Lipidure®-CM as a coating material, to ensure robust confinement of these highly adherent, migratory cells, enabling reliable tracking of phenotypic changes in large numbers of single cells and their progeny over time. Furthermore, the research team utilized CELLBANKER® cryopreservation media to preserve freshly isolated vSMCs, increasing the number of experiments that can be performed from a single tissue.

Key findings of this research demonstrate the remarkable benefits of CELLBANKER® cryopreservation media. Notably, 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%). Additionally, the researchers observed that CELLBANKER® 1&2 significantly outperformed DMSO-based protocols when assessing the growth of frozen primary cells. After just one week of culture, vSMCs preserved in CELLBANKER® 1 or 2 exhibited four to five times the cell growth compared to cells preserved in DMSO-based cryopreservation media.