3.1.2 Warming
Procedure
The uptake of glycerol by RBC is an active process [29]. For that
reason, at the time of glycerolization, the RBC and glycerol solution should be within a temperature range of 25–30
C. The
glycerol solution can be stored at room temperature. The RBCs
should be removed from the refrigerator and stored at room temperature for about 1 h prior to glycerolization.
1. Using a noncontact thermometer, check the temperature of
the RBC bag and the glycerol solution.
2. To warm the RBC bag or glycerol bottle, place them in a plastic
overwrap. Immerse the wrapped RBC bag or glycerol bottle in
a 37
C water bath until the desired temperature is reached.
Take care that no water enters the plastic overwrap bag.
3.1.3 Glycerolization
Procedure
Based on the weight of the RBC, the ACP215 automatically adds a
volume of 57% glycerol to the RBC to achieve a final glycerol
concentration of 40% (w/v). Power-on the ACP215. The
ACP215 automatically performs a series of internal system checks:
1. Select the glycerolization protocol.
2. Open the glycerolization disposable set and inspect it for visible
defects.
3. Install the disposable set according to the manufacturer’s
instructions [30].
4. Close the ratchet clamp on the spike connector tubing
segment.
5. Sterilely connect the RBC bag to the glycerolization disposable. Do not open the weld.
6. Load the blood pump tubing with the segment of tubing
between the two pump stops. Be sure that the blue pump
stop is toward the back of the machine. Secure the front and
back pump stops through their respective guides.
7. Load the pump outlet tubing into the blood line air detector
(BLAD).
8. Connect the draw pressure monitor (DPM) filter to the pressure sensor located on the front of the ACP215.
9. Remove the metal pull tab from the glycerol bottle, disinfect
the rubber stopper with an alcohol swab (70%), and insert the
spike end of the glycerolization disposable into the glycerol
bottle. Assure that the air vent on the spike is open to allow
air to flow into the bottle while it empties. If necessary, squeeze
the drip chamber to prime the chamber.
10. Suspend the glycerol bottle on the raised IV pole of the
ACP215.
Cryopreservation of Red Blood Cells
527
Procedure
The uptake of glycerol by RBC is an active process [29]. For that
reason, at the time of glycerolization, the RBC and glycerol solution should be within a temperature range of 25–30
C. The
glycerol solution can be stored at room temperature. The RBCs
should be removed from the refrigerator and stored at room temperature for about 1 h prior to glycerolization.
1. Using a noncontact thermometer, check the temperature of
the RBC bag and the glycerol solution.
2. To warm the RBC bag or glycerol bottle, place them in a plastic
overwrap. Immerse the wrapped RBC bag or glycerol bottle in
a 37
C water bath until the desired temperature is reached.
Take care that no water enters the plastic overwrap bag.
3.1.3 Glycerolization
Procedure
Based on the weight of the RBC, the ACP215 automatically adds a
volume of 57% glycerol to the RBC to achieve a final glycerol
concentration of 40% (w/v). Power-on the ACP215. The
ACP215 automatically performs a series of internal system checks:
1. Select the glycerolization protocol.
2. Open the glycerolization disposable set and inspect it for visible
defects.
3. Install the disposable set according to the manufacturer’s
instructions [30].
4. Close the ratchet clamp on the spike connector tubing
segment.
5. Sterilely connect the RBC bag to the glycerolization disposable. Do not open the weld.
6. Load the blood pump tubing with the segment of tubing
between the two pump stops. Be sure that the blue pump
stop is toward the back of the machine. Secure the front and
back pump stops through their respective guides.
7. Load the pump outlet tubing into the blood line air detector
(BLAD).
8. Connect the draw pressure monitor (DPM) filter to the pressure sensor located on the front of the ACP215.
9. Remove the metal pull tab from the glycerol bottle, disinfect
the rubber stopper with an alcohol swab (70%), and insert the
spike end of the glycerolization disposable into the glycerol
bottle. Assure that the air vent on the spike is open to allow
air to flow into the bottle while it empties. If necessary, squeeze
the drip chamber to prime the chamber.
10. Suspend the glycerol bottle on the raised IV pole of the
ACP215.
Cryopreservation of Red Blood Cells
527