16. When the deglycerolization procedure is completed, the
machine will beep and the following message will be displayed:
“Is the color of waste supernatant acceptable?” The color in the
supernatant in the tubing just before the line enters the calibrated line sensor, should be nearly colorless. Press YES if so or
NO if the color is not acceptable. Pressing the YES or NO key
causes the ACP215 to automatically resume operation. If NO
is chosen, it will be marked on the print out that the color was
not acceptable (see Note 7).
17. The printer will print out information related to the deglycerolization procedure.
18. Close all clamps on the lines of the disposable set to prevent
flowing of fluids.
19. Heat-seal the tubing between the RBC bag and the deglycerolization set and between the waste bag and the deglycerolization set. Label the RBC bag before disconnecting from the
ACP215.
20. Detach the RBC bag and waste bag from the rest of the
deglycerolization set.
21. Weigh the RBC bag and the waste bag and record the net
weights.
22. Sterilely connect a sample bag to the RBC bag, and transfer
about 5 mL of cell suspension for quality control.
23. Remove the disposable set from the machine and discard the
disposable set.
24. Store the deglycerolized RBC at 2–6
C until use.
3.3 Deglycerolization
of LGM Frozen Units
Although the ACP215 has been developed for processing RBCs
according to the HGM, it can also be used for deglycerolization of
LGM units [27]. The only adjustment needed in the deglycerolization procedure is the dilution of the initially added hypertonic salt
solution. While HGM frozen cells required a first dilution with 12%
NaCl, LGM cells are first diluted with 6% NaCl. Because the LGM
(de)glycerolization procedure is not performed in a functionally
closed system, post-thaw storage of these units is limited to 24 h:
1. Aseptically dilute the 12% NaCl solution 1:1 with water to
obtain a 6% NaCl solution.
2. Transfer the thawed LGM cells from the storage container to a
1000 mL transfer bag.
3. Perform the deglycerolization procedure as described above,
using the 6% NaCl solution instead of the 12% NaCl solution.
4. Upon the message “Thawed unit hematocrit,” use the
YES/UP or NO/DOWN arrows to adjust the setting to 30%
(LGM units are frozen without volume reduction).
532
Johan W. Lagerberg
machine will beep and the following message will be displayed:
“Is the color of waste supernatant acceptable?” The color in the
supernatant in the tubing just before the line enters the calibrated line sensor, should be nearly colorless. Press YES if so or
NO if the color is not acceptable. Pressing the YES or NO key
causes the ACP215 to automatically resume operation. If NO
is chosen, it will be marked on the print out that the color was
not acceptable (see Note 7).
17. The printer will print out information related to the deglycerolization procedure.
18. Close all clamps on the lines of the disposable set to prevent
flowing of fluids.
19. Heat-seal the tubing between the RBC bag and the deglycerolization set and between the waste bag and the deglycerolization set. Label the RBC bag before disconnecting from the
ACP215.
20. Detach the RBC bag and waste bag from the rest of the
deglycerolization set.
21. Weigh the RBC bag and the waste bag and record the net
weights.
22. Sterilely connect a sample bag to the RBC bag, and transfer
about 5 mL of cell suspension for quality control.
23. Remove the disposable set from the machine and discard the
disposable set.
24. Store the deglycerolized RBC at 2–6
C until use.
3.3 Deglycerolization
of LGM Frozen Units
Although the ACP215 has been developed for processing RBCs
according to the HGM, it can also be used for deglycerolization of
LGM units [27]. The only adjustment needed in the deglycerolization procedure is the dilution of the initially added hypertonic salt
solution. While HGM frozen cells required a first dilution with 12%
NaCl, LGM cells are first diluted with 6% NaCl. Because the LGM
(de)glycerolization procedure is not performed in a functionally
closed system, post-thaw storage of these units is limited to 24 h:
1. Aseptically dilute the 12% NaCl solution 1:1 with water to
obtain a 6% NaCl solution.
2. Transfer the thawed LGM cells from the storage container to a
1000 mL transfer bag.
3. Perform the deglycerolization procedure as described above,
using the 6% NaCl solution instead of the 12% NaCl solution.
4. Upon the message “Thawed unit hematocrit,” use the
YES/UP or NO/DOWN arrows to adjust the setting to 30%
(LGM units are frozen without volume reduction).
532
Johan W. Lagerberg