3.4 Quality Control
and Post-thaw Storage
The original RBC bag, the glycerolized RBC bag, the waste bag,
and the deglycerolized RBC bag are weight, and net weight is
recorded. The volume of the blood components is calculated
from the net weight divided by the specific gravity: 1.027 for
plasma, 1.100 for RBCs, 1.100 for 40% glycerol, and 1.006 for
SAGM. Using the hematocrit of the solution, the specific gravity of
RBC in additive solution is calculated from these values. For the
deglycerolization waste solution, a specific density of 1.04 is used.
The original RBC bag, the glycerolized RBC bag, and the deglycerolized RBC bag are sampled using sterilely connected sample
bags (see Note 8):
1. Analyze samples for hemoglobin concentration using HemoCue Hb analyzer and hematocrit using a microhematocrit
centrifuge (see Note 9).
2. Calculate total hemoglobin (g) as volume (mL) Â Hb concentration (g/dL)/100.
3. Calculate freeze recovery (%) as total Hb glycerolized RBC/total Hb original RBC Â 100.
4. Calculate freeze/thaw/wash recovery as total Hb deglycerolized RBC/total Hb original RBC Â 100.
5. A representative example of the calculations of a glycerolization/deglycerolization procedure is depicted in Table 1. In the
validation of the deglycerolization procedure, the efficacy of
the washing process should be determined. This can be done
by measuring the osmolarity of the supernatant of the deglycerolized product (see Note 10).
6. After deglycerolization, the RBCs should be stored at 2–6
C.
Since small changes in the glycerolization/deglycerolization
procedure can influence the post-thaw stability of the cells,
the post-thaw outdating time should be determined by each
institute individually (see Note 11).
4 Notes
1. The military blood bank (MBB) in the Netherlands holds a
stock of frozen (À80
C) RBCs (blood group O) for military
deployments. Their starting material is leukoreduced whole
blood. Whole blood units are leukoreduced after overnight
hold at ambient temperature. The units are processed and
frozen within 24 h after collection. Before glycerolization, the
plasma of the whole blood units is removed, and before freezing supernatant glycerol is removed [1, 2].
2. We use this for identification of the ACP215 used (AA, BB).
Cryopreservation of Red Blood Cells
533
and Post-thaw Storage
The original RBC bag, the glycerolized RBC bag, the waste bag,
and the deglycerolized RBC bag are weight, and net weight is
recorded. The volume of the blood components is calculated
from the net weight divided by the specific gravity: 1.027 for
plasma, 1.100 for RBCs, 1.100 for 40% glycerol, and 1.006 for
SAGM. Using the hematocrit of the solution, the specific gravity of
RBC in additive solution is calculated from these values. For the
deglycerolization waste solution, a specific density of 1.04 is used.
The original RBC bag, the glycerolized RBC bag, and the deglycerolized RBC bag are sampled using sterilely connected sample
bags (see Note 8):
1. Analyze samples for hemoglobin concentration using HemoCue Hb analyzer and hematocrit using a microhematocrit
centrifuge (see Note 9).
2. Calculate total hemoglobin (g) as volume (mL) Â Hb concentration (g/dL)/100.
3. Calculate freeze recovery (%) as total Hb glycerolized RBC/total Hb original RBC Â 100.
4. Calculate freeze/thaw/wash recovery as total Hb deglycerolized RBC/total Hb original RBC Â 100.
5. A representative example of the calculations of a glycerolization/deglycerolization procedure is depicted in Table 1. In the
validation of the deglycerolization procedure, the efficacy of
the washing process should be determined. This can be done
by measuring the osmolarity of the supernatant of the deglycerolized product (see Note 10).
6. After deglycerolization, the RBCs should be stored at 2–6
C.
Since small changes in the glycerolization/deglycerolization
procedure can influence the post-thaw stability of the cells,
the post-thaw outdating time should be determined by each
institute individually (see Note 11).
4 Notes
1. The military blood bank (MBB) in the Netherlands holds a
stock of frozen (À80
C) RBCs (blood group O) for military
deployments. Their starting material is leukoreduced whole
blood. Whole blood units are leukoreduced after overnight
hold at ambient temperature. The units are processed and
frozen within 24 h after collection. Before glycerolization, the
plasma of the whole blood units is removed, and before freezing supernatant glycerol is removed [1, 2].
2. We use this for identification of the ACP215 used (AA, BB).
Cryopreservation of Red Blood Cells
533