The Preservation of Human Red Blood Cell Agglutinogens in Liquid Nitrogen: Study of a Technic Suitable for Routine Blood Banking

Abstract

A simple method for the preservation of red blood cells in liquid nitrogen suitable for routine laboratory use is described. With this method, erythrocyte antigens retain their integrity for at least six months and, after thawing, remain active for at least two weeks in Alsever’s Solution. A panel of cells preserved in liquid nitrogen is as satisfactory as fresh cells in defining irregular antibodies encountered in patient sera. The advantages of this technic over other methods of red blood cell preservation is discussed.

Effect of red blood cell preservation by droplet freezing with non‐permeable cryoprotective agents in blood group antigen reactivity

“…The stability of RBC antigens after freezing in liquid nitrogen using CPAs, such as sucrose–dextrose and Alsever‐glucose solution, with few or no alterations in the antigenic pattern after thawing, was described decades ago (Huntsman et al ., , ; Bronson & McGinniss, ). These studies suggested that a panel of cells preserved in liquid nitrogen could define irregular antibodies in patient sera as precisely as could fresh cells.…”

“…This study aimed to evaluate the antigenic reactivity of fresh RBCs in comparison to erythrocytes preserved by droplet freezing in liquid nitrogen with two CPAs (S + D and PVP), using common blood group antigens with different molecular structures (Duffy, Lewis and Rh). Although it is not known whether the antigens evaluated are particularly susceptible to freezing and thawing, previous studies indicate that the alteration of Duffy, Lewis and Rh antigen reactivity may occur after RBC preservation both in suspension and frozen in liquid nitrogen using Alsever‐glucose solution or low glycerol method (Bronson & McGinniss, ; Myhre et al ., ; Allan et al ., ; Seo & Han, ).…”

“…These antigens were selected as indicators of antigenic stability since the loss of Duffy, Lewis and Rh antigens after RCB preservation in both fresh and frozen samples was reported by previous studies (Bronson & McGinniss, ; Myhre et al ., ; Allan et al ., ; Seo & Han, ).…”

“…Phenotyping tests for C and Fy b antigens were carried out before and 30 days after the RBC cryopreservation using anti‐C and anti‐Fy b antibodies in serial two‐fold dilutions in isotonic saline (Fig. c), in order to evaluate quantitatively the stability of the antigens (Bronson & McGinniss, ). The results were graded from 1+ to 4+ according to the intensity of agglutination.…”

“…Previous studies have shown that RBCs maintain their rheological properties, antigenic pattern and usability for serologic testing after freezing (Bronson & McGinniss, ; Pegg et al ., ; Henkelman et al ., ). Although the freezing of RBCs in glycerol is widely used by blood group reference laboratories (Smith, ; Högman et al ., ), the preservation in liquid nitrogen by droplet freezing, using non‐permeable CPAs such as polyvinylpyrrolidone (PVP) and sucrose–dextrose solution (S + D), has emerged as an effective alternative for the cryopreservation of small aliquots of erythrocytes for posterior use in serological tests (Schmid et al ., ).…”