3. With a RBC volume of about 170 mL after removal of supernatant, the volume of the original RBC storage bag (600 mL) is
large enough to hold the glycerolized cells (final volume about
570 mL). With larger RBCs, for instance, made from leukodepleted whole blood, the RBCs should be transferred to the 1 L
freezing bag before removal of the supernatant. After removal
of supernatant, the hematocrit of the suspension is very high
making transfer to another bag very hard.
4. Recently it has been shown that removal of supernatant glycerol before freezing is not necessary [32]. This not only simplifies the glycerolization procedure but may also reduce RBC
lesions by shortening the exposure to centrifugal force. Because
of the larger volume of the glycerolized cell suspension, omitting pre-freeze volume reduction could require larger cardboard boxes, storage containers, and therefore more storage
capacity. After thawing, because of the larger volume, the
deglycerolization procedure takes slightly longer but is still
efficient in removing all glycerol. Thawed units frozen according to the modified glycerolization procedure meet the quality
requirements of the European Council and AABB standards,
Table 1
Representative example of a freeze/thaw process
starting material
net weight RBC in SAGM (g)
319
density (g/mL)
1.064
volume starting material (mL)
300
Hb concentration (g/dL)
18.9
total Hb (g) starting material
56.7
glycerolization
net weight RBC after glycerolization
633
hematocrit (%)
34
volume frozen suspension (mL)
575
Hb concentration (g/dL)
9.7
total Hb frozen suspension (g)
55.8
freeze recovery (%)
99
deglycerolization procedure
waste fluid
nett weight bag waste fluid (g)
2012
volume waste fluid (mL)
1935
Hb concentration waste fluid (g/dL)
0.47
total Hb in waste fluid (g)
9.1
deglycerolized product
net weight RBC in SAGM (g)
300
hematocrit (%)
52
specific gravity (g/mL)
1.054
volume RBC in SAGM (mL)
284
Hb concentration RBC in SAGM (g/dL)
14.9
total Hb deglycerolized product
42.3
freeze/thaw/wash recovery (%)
75
Values marked with gray are measured values, and all other values
are calculated
534
Johan W. Lagerberg
large enough to hold the glycerolized cells (final volume about
570 mL). With larger RBCs, for instance, made from leukodepleted whole blood, the RBCs should be transferred to the 1 L
freezing bag before removal of the supernatant. After removal
of supernatant, the hematocrit of the suspension is very high
making transfer to another bag very hard.
4. Recently it has been shown that removal of supernatant glycerol before freezing is not necessary [32]. This not only simplifies the glycerolization procedure but may also reduce RBC
lesions by shortening the exposure to centrifugal force. Because
of the larger volume of the glycerolized cell suspension, omitting pre-freeze volume reduction could require larger cardboard boxes, storage containers, and therefore more storage
capacity. After thawing, because of the larger volume, the
deglycerolization procedure takes slightly longer but is still
efficient in removing all glycerol. Thawed units frozen according to the modified glycerolization procedure meet the quality
requirements of the European Council and AABB standards,
Table 1
Representative example of a freeze/thaw process
starting material
net weight RBC in SAGM (g)
319
density (g/mL)
1.064
volume starting material (mL)
300
Hb concentration (g/dL)
18.9
total Hb (g) starting material
56.7
glycerolization
net weight RBC after glycerolization
633
hematocrit (%)
34
volume frozen suspension (mL)
575
Hb concentration (g/dL)
9.7
total Hb frozen suspension (g)
55.8
freeze recovery (%)
99
deglycerolization procedure
waste fluid
nett weight bag waste fluid (g)
2012
volume waste fluid (mL)
1935
Hb concentration waste fluid (g/dL)
0.47
total Hb in waste fluid (g)
9.1
deglycerolized product
net weight RBC in SAGM (g)
300
hematocrit (%)
52
specific gravity (g/mL)
1.054
volume RBC in SAGM (mL)
284
Hb concentration RBC in SAGM (g/dL)
14.9
total Hb deglycerolized product
42.3
freeze/thaw/wash recovery (%)
75
Values marked with gray are measured values, and all other values
are calculated
534
Johan W. Lagerberg