“Control” Erythrocytes for Hemolysis Studies. ∗

Abstract

In several previous experiments the permeability of chicken erythrocytes at 37.5†C was studied over periods of many hours duration. It was noted that the time for hemolysis in glycerol of untreated cells became less after several hours (Hunter). 1 Since no attempt was made in these experiments to control bacterial contamination, it was thought that a part of the change in these control cells might have resulted from bacterial action. In another series of experiments, however, in which aseptic technics were followed it was also observed that the time for hemolysis of control cells decreased with lapse of time (Hunter and Larsh). 2 Results of this nature might have been predicted in view of previous experiments. Jacobs and Parpart 3 have shown that erythrocytes are quite sensitive to environmental changes while Harris 4 showed that under certain conditions of standing, erythrocytes lose potassium.

First a series of experiments were performed in which blood was allowed to stand at 37.5°C and time for hemolysis in glycerol was measured at varying intervals of time. Aseptic technic was observed throughout and tests were made for bacterial contamination as before (Hunter and Larsh). 2 Four stock suspensions were made. (1) Two cc of freshly drawn, heparinized chicken blood, (2) 1 cc of blood plus 1 cc of Ringer Locke, (3) 1 cc of cells plus 1 cc of plasma and (4) 1 cc of cells plus 1 cc of Ringer Locke. The results obtained using whole blood are shown in Fig. 1. Essentially similar results were obtained with all 4 solutions except the Ringer Locke appeared to exert a “protective effect” especially in (4). A similar series was run using blood which had been in the refrigerator for 27 hours. Hematocrit readings indicated that these cells had swollen although the initial hemolysis times and subsequent changes paralleled those obtained with the freshly drawn blood.

A second series of experiments involved hemolysis measurements, cell counts (Par-part), 5 and hematocrit determinations using an air turbine. These data are presented in Table I. It can be seen that with the passage of time at 37.5†C the cells swell as well as hemolyze more rapidly.

To determine whether the swelling could account for the change in rate of hemolysis, equal volumes of freshly drawn blood and 80% Ringer Locke (80 cc of Ringer Locke plus 20 cc of water) were mixed. At the end of 10 minutes the measurements were made. These data are presented in Table II.