Freezing in the Primary Polyvinylchloride Plastic Collection Bag: A New System for Preparing and Freezing Nonrejuvenated and Rejuvenated Red Blood Cells

Abstract

Red blood cells were stored at 4 C in the primary bag with an integrally attached empty transfer pack so that the red blood cells could be rejuvenated or not, as desired before glycerolization and freezing. The rejuvenation and glycerol solutions were added through ports in the system. After glycerolization, the red blood cells were concentrated by centrifugation to remove the supernatant glycerol before freezing with 40% w/v glycerol in the primary polyvinylchloride (PVC) plastic container at -80 C. After thawing, the red blood cells were washed using either the Haemonetics Blood Processor 115 or the IBM Blood Processor 2991-1 or 2991-2. In each system, 50 ml of 12% sodium chloride and 1.5 to 1.6 liters of 0.9% sodium chloride-0.2% glucose-25 meq/l disodium phosphate were used. Recovery of red blood cells in vitro was 91 per cent. After three days of postwash storage at 4 C, nonrejuvenated red blood cells had a mean 24-hour posttransfusion survival of 88 per cent, and outdated-rejuvenated red blood cells a value of 81 per cent. This new system is simpler and safer than methods previously used in this laboratory, and red blood cell recovery and 24-hour posttransfusion survivals were comparable or better.

An Experiment with Glycerol-Frozen Red Blood Cells Stored at –80°C for up to 37 Years

“…The glycerolized red cells were centrifuged, the supernatant glycerol solution was removed, and the glycerolized red cells in the 800-ml primary PVC plastic collection bag were placed in a rigid cardboard container for storage at –80°C [3, 4]. …”

“…Red cells frozen using the Cohn and Huggins methods were washed with 100–150 ml of 12% sodium chloride and 2 liters of 0.9% sodium chloride-0.2% glucose solution with a pH of 5.0 and an osmolality of 290 mosm/kg H 2 O (Baxter Healthcare, Deerfield, Ill.) as described by Kurtz et al [5]. Units frozen using the NBRL method were washed with 50 ml of 12% sodium chloride and 1.5 liters of 0.9% sodium chloride-0.2% glucose solution as described by Valeri et al [4]. The washed red cells were sampled following washing and after storage in the sodium chloride-glucose wash solution at 4°C for 24 h.…”

Appendices

“…The bag is centrifuged, the supernatant is expressed and the red cells then stored at −80°C. When needed, the unit is thawed in a waterbath at 37°C and the red cells washed free of glycerol, preferably with an automated cell washer (slightly modified from Valeri et al 1981).…”

Long-Term Storage and Preservation of Red Blood Cells,

“…Naval Blood Research Lab developed a minor, but important, change in the freezing/thawing/washing procedure. This change allowed the red cells to be frozen in the primary bag in which the cells had been drawn (Valeri et al 1981). The size of the primary bag for this procedure was increased from 600 to 800 ml.…”