2 Materials
2.1 Equipment
1. ACP215
automatic
cell
processor
(Haemonetics,
Braintree, MA).
2. Centrifuge for blood bags (e.g., Hettich Roto Silenta RP).
3. Centrifuge for tubes (e.g., Hettich Rotanta).
4. Freezer À80
C.
5. HemoCue plasma/low Hb analyzer.
6. Kocher clamp.
7. Manual plasma extractor.
8. Microhematocrit centrifuge.
9. Noncontact thermometer.
10. Scale.
11. Sterile connection device (Terumo).
12. Tube sealer (e.g., Fresenius CompoSeal Mobilea).
13. Vacuum bag sealer.
14. Water bath.
2.2 Reagents
1. Glycerol 57% solution (H-GLYC, S.A.L.F. S.p.A., Cenate
Sotto (Bergamo), Italy), bottle 500 mL. Contains per
100 mL: 57 g glycerol, 1.6 g sodium lactate, 30 mg monobasic
sodium phosphate (monohydrate), 124 g dibasic sodium phosphate (water free), pH 6.8.
2. 12% sodium chloride solution (SOL.IPERT.100, S.A.L.F.),
bag 100 mL.
3. Washing solution, 0.9% NaCl, 0.2% dextrose (SOL.FIS.2000,
S.A.L.F.), bag 2000 mL.
4. Red cell-additive solution SAG-M (Haemonetics, bag
300 mL).
5. Red blood cell concentrate, leukocyte depleted.
2.3 Disposables
1. Capillaries for hematocrit analysis.
2. Cardboard box (Cekumed, Ooltgensplaat, Netherlands).
3. Deglycerolization set (LN235, Haemonetics).
4. Freezing bag: 1000 mL PVC bag VSE 6002Z (MacoPharma,
Mouvaux, France).
5. Glycerolization set (LN225, Haemonetics).
6. Plastic overwrap bag.
7. Transfer bag 150 mL (e.g., P4159, Fresenius HemoCare,
Emmer-Compascuum, Netherlands).
8. Transfer bags 500 mL (e.g., P4162, Fresenius HemoCare).
Cryopreservation of Red Blood Cells
525