Freezing and storage of leukodepleted erythrocyte suspensions

Abstract

Studies on the frozen storage of human blood products have benefited veterinary transfusion medicine in recent years, but the long-term cryopreservation of canine red blood cells (RBCs) has not yet been thoroughly investigated. Further, no studies are available with respect to the frozen storage of leukocyte-depleted canine red blood cells (LD-RBCs). The objective of the current study was to investigate time-dependent effects of long-term frozen storage on leukocyte-depleted canine RBCs. Twelve healthy adult dogs meeting the criteria for blood transfusion were used in the study. Whole blood samples (450 ± 45 ml) collected from each dog were centrifuged for 5 min at 22 °C and 4200 × g in a cryogenic microcentrifuge and concentrated RBC (pRBC) suspensions were obtained. Leukocyte depletion was achieved by filtration (2.6 log 10 ). Then, the filtrated samples were prewashed three times in 0.9% NaCl solution and were allocated into three subgroups to be evaluated at three different time points (Day 0, Month 4 and Month 6). The samples for cryopreservation were subjected to glycerolisation and then stored at -80 °C for 4-and 6-month periods. At the end of this period pRBC units were thawed by manual agitation in a water bath maintained at 36-38 °C, centrifuged and then washed in a consecutive series of 12%, 1.6% and 0.9% of NaCl + 0.2 dextrose solutions. 2,3-Diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), supernatant haemoglobin (SupHb), sodium (Na + ) and potassium (K + ) levels, residual glycerol concentrations and haemograms of thawed and deglycerolised pRBC samples were evaluated together with those of Day 0. Sterility tests were performed on all samples for bacterial contamination. No statistically significant differences were noted except for Hct and SupHb levels. No bacterial contamination was noted in any of the samples on the basis of sterility tests. It was found that the described glycerolisation procedure could be a method of choice in the cryopreservation of leukocyte-depleted pRBCs (LD-pRBCs) since no negative effect was observed on the quality of the products and long-term frozen storage did not cause RBC destruction.

Cryopreservation of feline red blood cells in liquid nitrogen using glycerol and hydroxyethyl starch

“…[6][7][8][9][10][11][12] Canine cryopreservation techniques have also been successfully described in three previous reports using modifications of both automated and manual human protocols. [19][20][21][22] However, our findings are consistent with a recent study 23 that evaluated glycerol and HES as cryoprotectants for both canine and feline pRBCs. This study reported that neither HES nor glycerol were able to cryopreserve and recover feline RBCs successfully.…”

“…[9][10][11][12][13][15][16][17][18][19][20][21][22] In developing the protocol used in this study, we found that the materials and methods sections of the various publications had limited details related to each protocol, especially the manual techniques involved. [9][10][11][12][13][15][16][17][18][19][20][21][22] For instance, while Contreras et al described in detail the manual thawing and washing protocol for cryopreservation of canine pRBCs with glycerol, their protocol did not describe the method of agitation used when adding glycerol to the RBCs. 19 In addition, while Contreras et al used an automated RBC processor for the washing steps, Kim et al did not detail the washing steps in their protocol.…”

“…6–12 Canine cryopreservation techniques have also been successfully described in three previous reports using modifications of both automated and manual human protocols. 19–22…”

“…8,13,18 Investigations into frozen canine RBCs have shown successful cryopreservation for up to 2 months using both glycerol and HES. [18][19][20][21][22] Kim et al reported that 12.5% weight/weight (w/w) HES showed comparable cryoprotection of canine pRBCs to 20% glycerol based on hematologic and morphological parameters. 20 Cryopreserved canine pRBCs using 20% or 40% glycerol after thawing and washing, 19 and 11.5% (w/w) HES after thawing, 20 have been transfused into dogs with no transfusion reactions.…”

Leucorreducción Universal en Costa Rica: ¿Es realmente urgente?

“…Contáctenos: rev.med.ucr@gmail.com pacientes sometidos a procedimientos cardíacos se ha visto una disminución de las lesiones por reperfusión al reducir la activación de los neutrófilos, la producción de superóxido, complemento y leucotrienos después de la restauración del flujo sanguíneo [28,29 Los pacientes con enfermedad renal en etapa terminal experimentan barreras creadas por la aloinmunización. Esto se ejemplifica por el hecho de que aproximadamente el 30% de los candidatos a trasplantes renales en lista de espera están sensibilizados.…”