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 4x10^11 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. The IBM Blood Processor 2997 required the longest time for set-up and priming and processed 1.5 liters more donor blood per collection than the other instruments. The Fenwal CS-3000, which is a computer-controlled instrument, was the least time consuming. The donor suffered a significantly greater drop in platelet count after collection with the IBM Blood Processor 2997 (31%) than after collection with the other instruments (19%), and we were unable to account for this observation.
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. The platelets were frozen with a 6% cryoprotectant (DMSO) in a polyvinylchloride (PVC) plastic bag or a polyolefin plastic bag at -80°C in a mechanical freezer and stored for as long as 3 years. Some of the frozen platelets were transported in dry ice in polystyrene foam containers to determine whether they would be adversely affected by such treatment. Platelet recovery after freezing, thawing and washing was about 75%. In the healthy male volunteers, in vivo recovery of autologous platelets 1 - 2 h after transfusion was about 33%, and the life span was about 8 days. In the thrombocytopenic patients, in vivo recovery values were 50% of those from fresh platelets. The transfusion of previously frozen washed platelets reduced clinical bleeding in the thrombocytopenic patients with bleeding. There was no evidence of quality deterioration in platelets after storage at -80°C for at least 2 years, as determined from in vitro recovery and in vivo survival values, nor was there any adverse effect as a result of shipment of the frozen platelets in dry ice in polystyrene foam containers from one facility to another.
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 polyvinylchloride (PL-146) plastic bag with the Haemonetics blood processor V-50 or the Fenwal CS-3000 blood processor. The 600-ml PVC plastic bag was modified by sealing its vertical edges using radio frequency to form a narrow bag with a volume of approximately 200 ml. A 125-volume of diluted apheresis cellular residue was collected, and the mononuclear cells were isolated as follows: the diluted cellular residue was layered onto 75 ml of Ficoll- Hypaque with a specific gravity of 1.077 and was centrifuged at 260 g for 30 min at 22°C. The supernatant plasma was removed. The mononuclear cell layer was transferred to a sterile 600-ml transfer bag, and the cells were washed with saline. Of the 4.24 ± 0.9 x 10^9 mononuclear cells applied to the gradient, approximately 3.73 ± 0.8 x 10^9 cells were recovered. The recovered cells consisted of 77.3 ± 8% lymphocytes, 19.0 ± 7% monocytes, and 3.6 ± 3% granulocytes. There was no significant difference in tissue culture growth in the CFU-GEMM assay of mononuclear cells whether the plastic tube or the plastic bag system was used. Aerobic bactériologie cultures were negative. The PL-146 plastic bag system used in this study proved to be a significant aid in isolating mononuclear cells from plateletpheresis residue.
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