A Method for the Preservation of Human Blood Group Erythrocyte Antigens in Liquid Nitrogen for a Test Cell Panel¹

Abstract

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 cells 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.

A Modified Container for Liquid Nitrogen Preservation of an Erythrocyte Test Cell Panel1

“…The mean and standard deviation of 4 freezing and thawing curves on separate samples was used to determine an optimal freezing and thawing time. The results are shown in figure 1, compared with those obtained using steel cylinders [Mohn et al, 1970] During the initial 40 sec, the freezing curve A was nonlinear. No definite eutectic point was seen in the critical zone for erythrocyte freezing damage, from just below 0 to -50 °C.…”

“…Results obtained with a modified aluminum container for liquid nitrogen cryopreservation of red cells are presented. The optimal times and rates for red cell freezing and thawing are contrasted with those obtained using a stainless steel container.Previously [Mohn et al, 1970] we outlined a successful liquid nitrogen technic for small vomue erythrocyte cryopreservation and recovery. The thawed cells were useful in a test cell panel, as their blood group antigens were potent and stable for at least 7 days.…”

“…Their height was 60 mm (Cryogenic Equipment Company, Buckeystown, Md., USA). They were treated and capped as previously described [Mohn et al, 1970], The technic for cell cryopreservation and recovery was otherwise unaltered. …”

The H Specificities of Butanol Extracts of Human Erythrocytes

“…Red cells used as test cells in hemagglutination experi ments were recovered from the frozen state in the vapor phase of liquid nitrogen [3] and used within 24 h. The anti-A, anti-B, and anti-H sera were of human origin.…”

The H, HI, I and i Blood Group Antigens in Butanol Extracts of Rabbit Tissues

“…[3]. These red cells were used within 24 h of recovery from the frozen state as test cells in the inhibition of agglutination ex periments.…”