Cryopreservation of Equine Platelet-rich Plasma

Abstract

Platelet-Rich Plasma (PRP) therapy harnesses the healing properties of platelets to treat injuries and wounds. PRP provides growth factors and fibrinogen which clots and forms a scaffold to promote healing. In PRP processing, the patient blood is separated into fractions, and the component with platelets (either with (LPRP) or without (PRP) white blood cells) is used for therapy. This project aims to determine if 10% DMSO cryopreserves blood cells in PRP and LPRP stored at -80C or -140C and to measure associated protein concentrations. Our hypotheses were: PRP blood cells in 10% DMSO at -140C and -80C would be >80% viable, protein levels in PRP without DMSO would be higher than in PRP with DMSO, and protein levels in LPRP without DMSO would be lower than in LPRP with DMSO. Whole blood was processed to generate PRP and LPRP. Samples were stored overnight in 4C, -80C, and -140C with or without 10% DMSO. They were thawed at room temperature, cell viability was measured by a Live/dead assay, and protein concentration was determined using a Bradford assay. 10% DMSO improved cell viability in -80C and -140C, offering a potential method for PRP and LPRP cryopreservation. There was no statistical correlation between percent viability and protein concentration. Data suggest that protein levels are lower after cryopreservation with DMSO than without DMSO. Future directions include quantification of specific proteins within PRP and LPRP and increased storage time to determine cell viability and protein stability under these conditions.