I will be making my first conference talk at the above meeting tomorrow (January 9) during the Early Researchers’ Session. My talk is at 17.15.
Development of a novel human cell cryopreservation platform
Tim Morris, Andy Picken, Chris Hewitt, Karen Coopman
Centre for Biological Engineering, Loughborough University, Loughborough, UK
Cryopreservation a peripheral but vital process in the development of allogeneic cell-based products. With products making it to market, there is a need to improve the methods and protocols of preservation, to move away from DMSO for example, and to allow for a more regulated, characterised and efficient mechanism. Cryopreservation is a mature process in bacteria studies and in reproductive medicine, but limited for stem cell products. This project aims to improve this.
Early work has firstly focused on benchmarking DMSO, Glycerol and 1,2-propanediol (PrOH) as cryopreservants for hOST cells after freezing and 10 serial passages. Secondly the toxicity of DMSO has been analysed.
Results show that of the cryoprotectants studied DMSO is the most successful at retaining high cell viability immediately post-thaw. However, after 10 serial passages, all but those cells preserved in PrOH return to similar viability, metabolism and growth.
Experiments show that exposure to DMSO prior to freezing only reduces viability to ~80% after 2 hours, whereas post-freeze exposure can kill all cells after 2 hours. Cells exposed to DMSO, frozen and then analysed show a sort of middle ground in between the two other results, however these cells recover to >90% viability after one passage. This study is currently being expanded to include apoptosis and phenotype analysis.
Future work will be investigating the possibility of Trehalose and polymer loading as a new platform for cryopreservation.
Coopman, K., 2011. Large-Scale Compatible Methods for the Preservation of Human Embryonic Stem Cells: Current Perspectives. Biotechnology Progress, 27(6), p.1511-1521.
Keros, V. et al., 2005. Optimizing cryopreservation of human testicular tissue: comparison of protocols with glycerol, propanediol and dimethylsulphoxide as cryoprotectants. Human Reproduction, 20(6), p.1676-1687.
Lynch, A.L. et al., 2010. Biopolymer mediated trehalose uptake for enhanced erythrocyte cryosurvival. Biomaterials. 31(23), p.6096-6103.
Abstract v3.1.1, prepared for 23rd Annual ESACT-UK Meeting, Holywell Park, Loughborough University, 9th-10th January 2013