Chapter 26
Frozen Blood Reserves
Johan W. Lagerberg
Abstract
Frozen blood reserves are an important component in meeting blood needs. The idea behind a frozen
blood reserve is twofold: to freeze units of rare blood types for later use by patients with special transfusion
needs and for managing special transfusion circumstances. The permeating additive glycerol is used as a
cryoprotectant to protect red blood cells (RBCs) from freezing damage. The use of thawed RBCs has been
hampered by a 24-h outdating period due to the potential bacterial contamination when a functionally
open system is used for addition and removal of the glycerol. The introduction of an automated, functionally closed system for glycerolization and deglycerolization of RBCs improved the operational practice.
More importantly, the closed process allowed for extended shelf life of the thawed RBCs. In the current
chapter, a cryopreservation procedure for RBCs using a functionally closed processing system is described.
Key words Cryopreservation, Glycerol, Red blood cells, SAGM
1 Introduction
Cryopreservation is a valuable approach for managing an inventory of
rare RBC units to provide compatible blood for patients which are
negative for blood group antigens with a very high frequency in the
population (so-called public antigens). Frozen RBCs can also be used
for military deployments, which are characterized by logistical problems and unpredictable needs of blood components [1–3].
Glycerol is the most commonly used RBC cryoprotectant. The
use of glycerol as a cryoprotectant dates back to 1950 [4] after an
accidental discovery of the cryoprotective effect of glycerol the
previous year [5]. In the 1960s, it was observed that accelerating
the rate of freezing RBCs could significantly reduce the required
concentration of glycerol [6, 7]. After several modifications [8–11],
two methods employing glycerol at a concentration of 40% and
20% (w/v), referred to as high- (HGM) and low-glycerol method
(LGM), respectively, are still in use for freezing units of RBCs. In
the LGM, glycerol is added to a final concentration of approximately 20% (w/v). This method requires rapid freezing (i.e.,
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_26, © Springer Science+Business Media, LLC, part of Springer Nature 2021
523
Frozen Blood Reserves
Johan W. Lagerberg
Abstract
Frozen blood reserves are an important component in meeting blood needs. The idea behind a frozen
blood reserve is twofold: to freeze units of rare blood types for later use by patients with special transfusion
needs and for managing special transfusion circumstances. The permeating additive glycerol is used as a
cryoprotectant to protect red blood cells (RBCs) from freezing damage. The use of thawed RBCs has been
hampered by a 24-h outdating period due to the potential bacterial contamination when a functionally
open system is used for addition and removal of the glycerol. The introduction of an automated, functionally closed system for glycerolization and deglycerolization of RBCs improved the operational practice.
More importantly, the closed process allowed for extended shelf life of the thawed RBCs. In the current
chapter, a cryopreservation procedure for RBCs using a functionally closed processing system is described.
Key words Cryopreservation, Glycerol, Red blood cells, SAGM
1 Introduction
Cryopreservation is a valuable approach for managing an inventory of
rare RBC units to provide compatible blood for patients which are
negative for blood group antigens with a very high frequency in the
population (so-called public antigens). Frozen RBCs can also be used
for military deployments, which are characterized by logistical problems and unpredictable needs of blood components [1–3].
Glycerol is the most commonly used RBC cryoprotectant. The
use of glycerol as a cryoprotectant dates back to 1950 [4] after an
accidental discovery of the cryoprotective effect of glycerol the
previous year [5]. In the 1960s, it was observed that accelerating
the rate of freezing RBCs could significantly reduce the required
concentration of glycerol [6, 7]. After several modifications [8–11],
two methods employing glycerol at a concentration of 40% and
20% (w/v), referred to as high- (HGM) and low-glycerol method
(LGM), respectively, are still in use for freezing units of RBCs. In
the LGM, glycerol is added to a final concentration of approximately 20% (w/v). This method requires rapid freezing (i.e.,
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_26, © Springer Science+Business Media, LLC, part of Springer Nature 2021
523