Leukocyte preservation. I. Phagocytic stimulation of the hexose monophosphate shunt as a measure of cell viability.

Abstract

Abstract Phagocytic stimulation of the hexose monophosphate shunt (HMS) was employed to quantitate human phagocyte viability. At leukocyte concentrations below 5,000 per cubic millimeter, and with an excess of latex particles, the 14 CO 2 derived from glucose-1- 14 C was directly proportional to the number of cells. When the number of particles was reduced and the cell count maintained constant, the radioactivity was proportional to the number of particles ingested. Leukocytes separated under sterile conditions were stored at 25 ° C. and at 4 ° C. in Hanks' balanced salt solution containing 30 per cent serum. The stimulated HMS activity of the cells stored at 25 ° C. decreased 50 per cent in 36 hours while the HMS activity in the cells stored at 4 ° C. decreased 50 per cent in 60 hours. Freezing leukocytes to −80 ° C. without a cryopreservative resulted in a loss of all HMS activity. Cells frozen to this temperature in 15 per cent dimethylsulfoxide, however, retained approximately 25 per cent of their prefreezing stimulated activity.