Simplification of the Methods for Adding and Removing Glycerol during Freeze-Preservation of Human Red Blood Cells with the High or Low Glycerol Methods: Biochemical Modification Prior to Freezing.

摘要

Simple methods have been developed for adding and removing glycerol during freeze-preservation with 20 per cent W/V glycerol at -150 C, or with 40 per cent W/V glycerol at -80 C. A one-step method with a 35 per cent W/V glycerol solution is used to prepare 20 per cent W/V glycerolized red blood cells, and a two-step method with a 57 per cent W/V glycerol solution is used to prepare 40 per cent W/V glycerolized red blood cells. The systems for washing glycerolized red blood cells have been simplified. This method consists of dilution of the thawed glycerolized red blood cells prior to recovery, followed by on-line dilution of these red blood cells with wash solutions during continuous flow centrifugation. This can be done in any of three commercially available washing systems, and they all use the same sodium chloride solutions. For the 40 per cent W/V glycerolized red blood cells, this process takes about 30 minutes and uses 2.2 to 3.2 liters of the sodium chloride solutions, whereas the 20 per cent W/V glycerolized red blood cells can be processed in about 20 minutes using 1.5 to 2.5 liters.