Isolation of mononuclear leukocytes in a plastic bag system using Ficoll-Hypaque.

Mononuclear cells, present in bone marrow and peripheral blood, have been isolated from red cells and granulocytes using a ficoll-hypaque density centrifugation process. Cells isolated by this process which uses centrifuge tubes may become contaminated. In 19 studies in our laboratory we used Ficoll-Hypaque treatment to isolate mononuclear cells from cellular residues obtained during plateletpheresis using a modified 600-ml polyvinyl-chloride (PL-146) plastic bag with the Haemonetics blood processor V-50 or the Fenwal CS-3000 blood processor.

Cryopreservation of human platelets using 6% dimethyl sulfoxide and storage at -80 degrees C. Effects of 2 years of frozen storage at -80 degrees C and transportation in dry ice.

Platelet studies were done in healthy male volunteers and in thrombocytopenic patients. Some of the platelets used in the study were isolated by mechanical apheresis using either the Haemonetics blood processor 30, the IBM blood processor 2997 or the Fenwal CS-3000 blood processor before freezing. Other platelets were isolated from individual units of whole blood and pooled before freezing.

Plateletpheresis experience with the Haemonetics Blood Processor 30, the IBM Blood Processor 2997, and the Fenwal CS-3000 Blood Processor.

Comparisons were made of the apheresis instruments Haemonetics Blood Processor 30, IBM Blood Processor 2997 and Fenwal CS-3000, for collection of platelets from normal donors. With each instrument the mean recovery was at least 4 x 10(11) platelets per procedure, and each instrument afforded a safe and reliable collection. The Haemonetics Blood Processor gave the lowest recovery of platelets per minute per procedure.

Factors that influence the process of 51chromium labeling of human granulocytes isolated from blood by counterflow centrifugation.

The uptake of 51chromium by human granulocytes isolated from blood by counterflow centrifugation is influenced by the composition of the suspension medium, the incubation period, the temperature and the amount of 51chromium used. Counterflow centrifugation-elutriation isolated approximately 98 per cent of the granulocytes from the blood. Acid-citrate-dextrose (ACD) plasma, citrate-phosphate-dextrose (CPD) plasma and heparinized plasma as resuspension media produced similar labeling results, whereas phosphate-buffered saline resulted in three times greater uptake in 51chromium.