Proposal of a New Standardized Freeze-Thawing Technical Protocol for Leucocyte-Poor Platelet-Rich Plasma Preparation and Cryopreservation

A human platelet-rich plasma (PRP) concentrate can be defined as a preparation of autologous human plasma with increased platelet concentration produced by centrifugation of a larger volume of a patient’s own blood. Platelets contain a plethora of growth factors in their α-granules that are concentrated through the centrifugation process in order to then be injected in supraphysiologic amounts to an injury site with the final aim of augmenting the natural healing process. Preparations of PRP concentrates can be further classified as leucocyte-rich (LR-PRP), defined as having a leucocyte concentration above baseline, and leucocyte-poor (LP-PRP), defined as having a leucocyte concentration below baseline. Although many preclinical and clinical trials have shown the ability of leucocyte-poor PRP concentrates to significantly improve symptomatic mild to moderate hip and/or knee osteoarthritis, to date there is no consensus on the optimal way of obtaining PRP preparations, specifically with respect to the concentration of blood components. In this technical report, we describe a new standardized freeze-thawing technique for leucocyte-poor PRP preparation and cryopreservation, which has been shown to be superior to currently available techniques based solely on centrifugation. By describing this technical protocol, which we have been using on a daily basis in the setting of a Regenerative Medicine Outpatient Clinic in a European tertiary university hospital center, we aim to contribute to a future consensus on the optimal way of obtaining and preserving leucocyte-poor PRP concentrates.