Platelet cryopreservation using a reduced dimethyl sulfoxide concentration and second-messenger effectors as cryopreserving solution.
Cryopreservation of platelets is of great interest since it could extend to years the shelf life of therapeutic platelet concentrates (PCs) and facilitate stockpiling and inventory control in blood banking. We have compared the cryopreservation of PCs by the standard method using 6% Me(2)SO as cryoprotectant with the method of freezing employing low concentrations of Me(2)SO (2%) plus ThromboSol, a mixture of second-messenger effectors that protect platelets from cold damage. PC pools were treated either with 6% Me(2)SO or with ThromboSol and 2% Me(2)SO and then placed directly in a -80 degrees C freezer or in the vapor phase of a liquid nitrogen freezer (-120 degrees C). After storage for 1 week or for 3 months, samples were removed, thawed, and analyzed. Measurements included cell recovery, biochemical parameters, membrane glycoproteins (GPs), platelet aggregation, and binding of radiolabeled von Willebrand factor (vWF) and fibrinogen. PCs cryopreserved with ThromboSol and 2% Me(2)SO displayed a platelet recovery (90%) equivalent to those frozen with 6% Me(2)SO. Following either cryopreservation procedure, platelets showed increased surface expression of P-selectin and moderate loss of GP Ibalpha in comparison to fresh platelets. The aggregatory response to ristocetin and the binding of vWF were similar in platelets frozen by either procedure. Finally, both methods promoted comparable impairment of the reactivity of platelets to thrombin, aggregation and binding of fibrinogen and vWF, compared to that of fresh platelets. In summary, cryopreservation of PCs using reduced Me(2)SO concentration and ThromboSol yields platelets with in vitro functional characteristics equivalent to those of cells frozen with the conventional method using 6% Me(2)SO.
Lozano
M L
ML
Unit of Hematology, Hemotherapy and Clinical Oncology, School of Medicine, Centro Regional de Hemodonación, Murcia, 30003, Spain.
Rivera
J
J
Corral
J
J
Gonzalez-Conejero
R
R
Vicente
V
V
D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D001793:Blood Preservation / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D005069:Evaluation Studies as Topic; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D010974:Platelet Aggregation / Q000187:drug effects; D010976:Platelet Count; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D012310:Ristocetin / Q000494:pharmacology; D015290:Second Messenger Systems; D012996:Solutions; D014841:von Willebrand Factor / Q000378:metabolism
D003451:Cryoprotective Agents; D010980:Platelet Membrane Glycoproteins; D012996:Solutions; D014841:von Willebrand Factor; D012310:Ristocetin; D005340:Fibrinogen; D004121:Dimethyl Sulfoxide