Methods of RBC Cryopreservation Consideration Highconcentration Glycerol Lowconcentration

Methods of RBC Cryopreservation Consideration High-concentration Glycerol Low-concentration Glycerol Final glycerol concentration(wt/vol) 40% 20% Freezing temperature -65 C -120 C Mechanical Liquid nitrogen Dry ice Liquid nitrogen Yes No 94 -99 95 Type of freezer Shipping Deglycerolyzing equipment required WBC removal (%)

Materials & Methods • 대상 Fresh packed RBCs for in vitro evaluation : 10 unit Autologous RBC for transfusion : 4 unit • 방법 High-concentration glycerol method

Freezing

Warming

Transfer RBCs to 1000 m. L bag

Mix RBCs withglycerolyte sol. Glycerolyte 57 solution

Balancing

Transfer the supernatant from 1000 m. L bag to packed RBC bag after centrifugation

The color of the supernatant

After transfer of the supernatant

Remove the packed RBC bag

Transfer the glycerolyzed RBC from 1000 m. L bag to 600 m. L bag Glycerolized RBC in canister

Thawing

Automatic Cell Washer Control Panel COBE 2991

Option for washing the thawed RBC

Add washing solution to the RBC bag 12 % Na. Cl processing solution

Transfer the mixed RBC with washing sol. to the centrifugal bag

Processing

Last washing 0. 2% dextrose and 0. 9% Na. Cl processing solution

The thawed-washed RBC

Results Parameters Recovery rate (>80%) S. G. (<1. 3384) K+ (<1. 5 m. Eq/L) Mean ± SD Range 72. 9 ± 8. 2 61. 4 -88. 9 1. 006 ± 0. 001 1. 005 – 1. 011 1. 17 ± 0. 42 0. 8 -2. 0 Hb ATP (3. 5 ± 0. 9 umol/g) 3. 5 ± 0. 9 Hb 2, 3 -DPG (12. 5± 5. 3 umol/g) 12. 5 ± 5. 3 Simulated transfusion (<30 mg/d. L) 2. 65 ± 1. 18 Blood culture No grwoth 1. 1 -4. 6

Autologous infusion • 4 normal healthy donor • Hct 43. 0%, • Vol : 220 (135 -300) m. L After transfusion • no Hemoglobinuria • Serum-haptoglobin 96. 0± 40. 8 mg/d. L • no adverse sign or symptom