連続遠沈法による解凍赤血球の検討

Abstract

1. Objective and ProcedureFresh red blood cells are to be frozen either slowly or rapidly with glycerol for preservation as viable cells in a frozen medium. From the viewpoint of deglycerolizing such preserved viable erythrocytes there have been introduced three different techniques, viz., the reversible agglomeration (Huggins' method), the batch washing and the continuous centrifuge procedure.We conducted a study attempting the evaluation of the continuous centrifuge technique for deglycerolizing preserved red cells, using a Hemonetics Model 15 apparatus, in comparison with Huggins' method as to effects on osmotic pressure, free Hg concentration, erythrocytic resistance, electrolyte balance, 2, 3-DPG and ATP levels and recovery rate of viable cells.2. ResultsAs compared with Huggins' procedure, the continuous centrifuge method produced rather inconsistent results for which the rates of red cell and washing medium injection appear to have played an important role. In case of the washing medium being introduced into the apparatus before erythrocytes it would be probable that sufficient washing of red cells could hardly be accomplished. The failure in obtaining constantly optimal osmotic pressure, erythrocyte resistance and electrolyte balance with the subject procedure has thus been conjectured to be due to insufficient washing of preserved cells.3. ConclusionThe continuous centrifuge seems to be valuable method provided it be performed attentively insomuch as the glycerol concentration employed is fairly low and as electrolyte solutions are used for washing the cells.