BACKGROUND: The cryopreservation of red blood cells (RBCs) has not ever been applied to the clinical services in Korea. The aim of this study was designed to supply the frozen-thawed RBCs as a routine service through estimation of efficiency and safety after freezing, thawing and washing. METHODS: Fifteen fresh packed RBCs were frozen with 40 percent(wt/vol) glycerol. After frozen storage at -70degrees for at least one month, the RBCs were thawed and washed in the COBE 2991 blood cell processor. We measured the blood cell count, RBC recovery rate, K+, LDH, specific gravity, osmolarity, and the percentage of hemolysis in the supernatant after deglycerolization. Autologous transfusions were done to the four voluntary donors with deglycerolized autologous blood for clinical assessment. RESULTS: The freeze-thaw-wash recovery rate of RBC was 76.8+/-10.0%, which is not enough to pass the AABB standard. But the recovery rate was increased up to 87.0+/-2.1% with the 4 stepwise predilution technique. The supernatant plasma specific gravity, osmolarity, and K+ were 1.006+/-0.001, 292+/-3 mOsm/KgH20, and 1.1+/-0.2mEq/L, respectively. The Hb ATP and 2,3-DPG were 3.6+/-0.8nmol/g and 13.4+/-4.5nmol/g. In simulated study, the free hemoglobin was 2.8+/-1.1mg/dL and 0.4+/-0.2% of total hemoglobin. In four autologous transfusion cases, plasma haptoglobin level was 96.0+/-40.8 mg/dL (reference range 30~200 mg/dL) and urine hemoglobin was not observed after 2~6 hours later after transfusion. CONCLUSION: The results of this study indicated that technical experiences for freezing, thawing and washing were established for clinical use of frozen RBCs in Korea.
2,3-Diphosphoglycerate ; Adenosine Triphosphate ; Blood Cell Count ; Blood Cells* ; Cryopreservation ; Erythrocytes* ; Freezing ; Glycerol ; Haptoglobins ; Hemolysis ; Humans ; Korea ; Osmolar Concentration ; Plasma ; Specific Gravity ; Tissue Donors

PURPOSE: The needs of stem cell transplantation are increasing in Korea under the circumstance of medical insurance coverage since 1996. But allogeneic bone marrow transplants have the limitation in acquisition of matched donor because of small number of siblings in modern society. Although unrelated stem cell transplantations were made possible in Korea with the establishment of Korean Marrow Donor Program, less than one third of patients who need stem cell transplantation are seeking the matched donors through this program and it still takes too much time and efforts. In this clinical setting, cord blood offers an exciting alternative to traditional sources of stem cells. To implement the widespread clinical use of cord blood stem cells, it is necessary to establish the cord blood bank from which stem cell specimens are obtained. METHODS: To establish the cord blood bank in Korea, we started to develop the proper methods to obtain the cord bloods. Various ways of collecting the cord bloods, separation of red blood cells and storage methods were tried and the standard recommendations were made. RESULTS: Mean volume of collected umbilical cord blood was above 70mL by open or closed method in vaginal delivery and cesarean section. Among the RBC depletion methods by using 3% gelatin, Ficoll-Hypaque, or modified Ficoll-Hypaque method, 3% gelatin sedimentation showed the best result in viability and number of mononuclear cells(MNC), CD34+ cell, and day 14 CFU-GM. The recovery rate after freezing-thawing MNC was higher in the group of using 10% DMSO and programmed freezer starting from 4degrees C than in the group of using mixed with DMSO and hydroxyethyl starch and without programmed freezer. Nine patients underwent cord blood transplantation until December 1998. Among them, 6 patients showed the long-term successful engraftment. CONCLUSION: We have developed simple, efficient, and reproducible methods for collection, processing, and storage of cord blood to establish the cord blood bank. There are some administrative issues including the legal and financial problems that possibly will be resolved by the support of the government and the medical team.
Bone Marrow ; Cesarean Section ; Dimethyl Sulfoxide ; Erythrocytes ; Female ; Fetal Blood* ; Gelatin ; Granulocyte-Macrophage Progenitor Cells ; Humans ; Insurance Coverage ; Korea* ; Pregnancy ; Siblings ; Starch ; Stem Cell Transplantation ; Stem Cells* ; Tissue Donors