Recovery of Glycerolized Red Blood Cells Frozen in Liquid Nitrogen

Abstract

Human red blood cells were frozen at a rapid rate in the presence of approximately 20 per cent w/v glycerol and an osmotic supporting agent, and were stored in liquid nitrogen vapor at approximately -150 C for at least two weeks before thawing, washing, and resuspension. The recovery of red blood celb upon thawing was dependent on the type of freezing container wed and the volume of red blood cells frozen. An additional loss of red blood celb during postthaw washing was related to the volume of red blood cells frozen, but was not related to the type of osmotic supporting agent used during the wash. The total amount of aupanatant hemoglobin in the washed unit was inversely related to the recovery of the red blood cells upon thawing. For adequate recovery of preserved red blood cells, both the geometry of the container in which the cells had been frozen, and the capacity of the reusable bowl-washing system employed for removing the glycerol, limited the m m of glycerolized red blood celh to less than 600 g.

A method for the separation of erythrocytes on the basis of size using counterflow centrifugation

“…EGOT levels were picked to study as an age-related enzyme because of the close relationship between EGOT and the age of an erythrocyte, as reported by Bartos and Desforges [ 141, and the availability of a highly quantitative assay for it in our laboratory. Hemoglobin was measured using the cyanmethemoglobin technique as previously described [22]. EGOT activity was assayed by measuring the conversion of NADH to NAD at 340 nm in the presence of oxaloacetic acid.…”