Tracey R. Turner

A quality monitoring program for red blood cell components: in vitro quality indicators before and after implementation of semiautomated processing

Canadian Blood Services has been conducting quality monitoring of red blood cell (RBC) components since 2005, a period spanning the implementation of semiautomated component production. The aim was to compare the quality of RBC components produced before and after this production method change.

Small molecule ice recrystallization inhibitors enable freezing of human red blood cells with reduced glycerol concentrations.

In North America, red blood cells (RBCs) are cryopreserved in a clinical setting using high glycerol concentrations (40% w/v) with slow cooling rates (~1°C/min) prior to storage at −80°C, while European protocols use reduced glycerol concentrations with rapid freezing rates. After thawing and prior to transfusion, glycerol must be removed to avoid intravascular hemolysis. This is a time consuming process requiring specialized equipment. Small molecule ice recrystallization inhibitors (IRIs) such as β-PMP-Glc and β-pBrPh-Glc have the ability to prevent ice recrystallization, a process that contributes to cellular injury and decreased cell viability after cryopreservation. Herein, we report that addition of 110 mM β-PMP-Glc or 30 mM β-pBrPh-Glc to a 15% glycerol solution increases post-thaw RBC integrity by 30-50% using slow cooling rates and emphasize the potential of small molecule IRIs for the preservation of cells.

Evaluating the 4‐hour and 30‐minute rules: effects of room temperature exposure on red blood cell quality and bacterial growth

BACKGROUND: A 30‐minute rule was established to limit red blood cell (RBC) exposure to uncontrolled temperatures during storage and transportation. Also, RBC units issued for transfusion should not remain at room temperature (RT) for more than 4 hours (4‐hour rule). This study was aimed at determining if single or multiple RT exposures affect RBC quality and/or promote bacterial growth.

Quality of red blood cells washed using an automated cell processor with and without irradiation

Sterile washing of red blood cells (RBCs) and use of an additive solution permits longer postwash storage. The effect of irradiation during this extended storage time is unclear.

Small molecule ice recrystallization inhibitors mitigate red blood cell lysis during freezing, transient warming and thawing.

During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to <1% prior to transfusion thus limiting the use of frozen RBC units in emergency situations. We have identified several low molecular mass ice recrystallization inhibitors (IRIs) that are effective cryoprotectants for human RBCs, resulting in 70–80% intact RBCs using only 15% glycerol and slow freezing rates. These compounds are capable of reducing the average ice crystal size of extracellular ice relative to a 15% glycerol control validating the positive correlation between a reduction in ice crystal size and increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming.

Quality of red blood cells washed using a second wash sequence on an automated cell processor

Washed red blood cells (RBCs) are indicated for immunoglobulin (Ig)A‐deficient recipients when RBCs from IgA‐deficient donors are not available. Canadian Blood Services recently began using the automated ACP 215 cell processor (Haemonetics Corporation) for RBC washing, and its suitability to produce IgA‐deficient RBCs was investigated.

O-Aryl-Glycoside Ice Recrystallization Inhibitors as Novel Cryoprotectants: A Structure–Function Study

Low-molecular-weight ice recrystallization inhibitors (IRIs) are ideal cryoprotectants that control the growth of ice and mitigate cell damage during freezing. Herein, we describe a detailed study correlating the ice recrystallization inhibition activity and the cryopreservation ability with the structure of O-aryl-glycosides. Many effective IRIs are efficient cryoadditives for the freezing of red blood cells (RBCs). One effective cryoadditive did not inhibit ice recrystallization but instead inhibited ice nucleation, demonstrating the significance of inhibiting both processes and illustrating the importance of this emerging class of cryoprotectants.

From Development to Implementation: Adjusting the Hematocrit of Deglycerolized Red Cell Concentrates to meet Regulatory Standards

Background: Before transfusion, thawed frozen red cell concentrates (RCCs) must be deglycerolized. In order to ensure that these products meet regulatory standards for hematocrit, an approach to manipulate hematocrit post deglycerolization was developed and implemented. Methods: Glycerolized and frozen RCCs were thawed and deglycerolized using the COBE 2991 cell processor, and the final products hematocrit was adjusted by addition of various volumes of 0.9% saline / 0.2% dextrose. The in vitro quality of RCCs (hematocrit, hemolysis, hemoglobin content, volume, recovery, ATP, supernatant potassium, and others) were compared to Canadian Standards Association (CSA) and other standards for deglycerolized RCCs. Results: Addition of saline/dextrose re-suspension solution in a range of 65-90 g post deglycerolization led to acceptable hematocrits. In the pilot study, this approach resulted in RCCs meeting all CSA standards for deglycerolized RCCs, with stimulation of RBC metabolism demonstrated by increased ATP concentration. In the validation phase, results were similar, although the CSA hemolysis standard was not met. Pre- and post-implementation data confirmed that manipulated RCCs met CSA hematocrit standards. Conclusion: This process was implemented at Canadian Blood Services to provide deglycerolized RCCs that meet the CSA hematocrit standard. However, pre- and post-implementation data reveal that this deglycerolization process is not sufficient to have RCCs consistently meet hemolysis standards.