A Method for the Preservation of Human Blood Group Erythrocyte Antigens in Liquid Nitrogen for a Test Cell Panel

Abstract

A modification of Krijnen’s cryopreservation method, using low intracellular concentrations of 17.5% glycerol and extracellular 4% sorbitol as additives, has been developed for liquid nitrogen freezing of small volumes of erythrocytes for a test cell panel. A technique for their subsequent successful storage in sterile, dextrose-electrolyte maintenance solution (CP-2) at 4°C after recovery is presented. By these procedures, the mean recovery of red ceils after lytic losses from freezing and thawing injury was 94.1%. The manipulations involved in transferring, mixing and resuspending the red cells removed an additional 11.1-16.2%, resulting in an overall recovery of 80.5% for serologic tests. Less than 3% of the cells exhibited cumulative lysis at seven days maintenance in CP-2 solution at 4°C, and the blood group antigens examined remained as potent as those in sterile, fresh ACD blood.

Blood Group Antigens I, i, H and HI on Monkey, Rabbit and Guinea Pig Erythrocytes

“…Whole blood from rabbits, monkeys, and guinea pigs was obtained by cardiac puncture and mixed with ACD solution, PHS formula B. All blood samples were maintained aseptically and were used up to 14 days following collection or were stored as frozen red cells in the vapor phase of liquid nitrogen at -150" according to the method of Mohn, Bowman and Cunningham (3).…”

An Inherited Blood Group A Variant in the Finnish Population

“…samples was kept at 4°C for examination within 5 days after receipt, and was always handled with sterile precautions whenever portions were removed. The red cells from the second ACD sample were frozen in liquid nitrogen [16] for later use. The serum was removed, with sterile precautions, from the clotted specimens after centrifugation, and frozen at -25 "C in small aliquots.…”