Human granulocytes were isolated from blood either by counterflow centrifugation in a Beckman JE-6 rotor or by sedimentation of the red blood cells with dextran and centrifugation of the granulocyte-rich plasma. The stability of the granulocytes was assessed during storage in the liquid state by measurements of granulocyte loss, volume distribution characteristics, and ability to produce fluorescein in their cytoplasm and to exclude ethidium bromide from their nuclei. After storage at 4 C for 2 days in phosphate-buffered saline with a pH of 7.1 containing 5 g/dl human albumin + 0.46 g/dl dextrose or 1 g/dl Physiogel + 0.46 g/dl dextrose, the granulocytes were adequately preserved from the in vitro measurements.

Blood Preservation↗

PubMedDate not supplied

In vitro studies of cryopreserved baboon granulocytes.

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Animals↗

PubMedDate not supplied

Isolation and cryopreservation of monocytes from plateletapheresis cellular residues.

Human monocytes were isolated from the cellular residues remaining after plateletapheresis of donors using an automated blood cell processor. Mononuclear cells were obtained with density gradients and separated from lymphocytes by stepwise elutriation. The isolated cells were frozen using extracellular hydroxyethyl starch and intracellular dimethylsulfoxide. In three procedures, approximately 1 X 10(9) monocytes were obtained. Ninety-nine percent of isolated monocytes were viable in a fluorescein diacetate (FDA)-ethidium bromide (EB) test. Myeloperoxidase-positive cells were 95 percent and 90 percent in the two chambers. Ninety-four percent of monocytes ingested five or more opsonized polycyclic hydrocarbon particles and 95 percent ingested one or more ethidium-treated zymosan particles. After storage in liquid nitrogen for up to 9 weeks, 99 percent of the cells were recovered after thawing. Of these, 95 percent were myeloperoxidase-positive, 94 percent showed intact membranes in the FDA-EB test, 95 percent ingested five or more opsonized polycyclic hydrocarbon particles, and 96 percent ingested one or more ethidium-treated zymosan particles. These results demonstrate the utility of elutriation as a means to isolate large numbers of monocytes. The isolated cells can be cryogenically preserved for at least 2 months with small loss of phagocytic function.

Blood Preservation↗