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A Method for Freezing and Washing Red Blood Cells Using a High Glycerol Concentration
Author(s) -
Meryman H. T.,
Hornblower M.
Publication year - 1972
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.1972.tb00001.x
Subject(s) - glycerol , tonicity , hemolysis , dilution , chromatography , chemistry , isotonic solutions , isotonic , lysis , sodium , cryopreservation , osmotic pressure , red blood cell , biochemistry , biology , medicine , immunology , embryo , physics , organic chemistry , microbiology and biotechnology , thermodynamics
A modification of previously reported methods for freezing red blood cells with a high concentration of glycerol is described. The limits of osmotic tolerance of human red blood cells range from one‐half to four times isotonic. The introduction and removal of glycerol creates transient osmotic gradients that can exceed these limits. It is shown that much of the cell lysis following thawing and during washing can be related to hypertonic injury incurred during the glycerolizing procedure. Refinements of this portion of the process produce significant gains in recovery. Problems associated with washing out a high concentration of glycerol by centrifugal means can be alleviated by a preliminary dilution with hypertonic sodium chloride which serves also to reduce cell volume and increase cell specific gravity improving sedimentation characteristics and affording more latitude for the hypotonic stresses associated with glycerol removal. Careful attenion to the osmotic limits during both glycerolizing and washing results in a process in which hemolysis from freezing, thawing and washing can be reduced to less than three per cent.