Continuous‐flow centrifugation of blood from normal donors with the NCI‐IBM Continuous Flow Blood Cell Separator can be used safely and effectively to collect large quantities of normal granulocytes, lymphocytes, and platelets. Comparison with single‐unit leukapheresis demonstrated a threefold greater collection of granulocytes and a fivefold greater collection of lymphocytes per hour of leukapheresis with continuous flow centrifugation. Granulocyte collections can be enhanced by giving the donors etiocholanolone or cortisone prior to leukapheresis. Plateletpheresis can be combined efficiently with granulocyte collction during continuous‐flow centrifugation, but it offers no particular advantage over platelet collection by standard plasmapheresis technics.
A closed continuous flow centrifuge (NCI-IBM Blood Cell Separator) was utilized to collect large quantities of leukocytes from donors with chronic myelocytic leukemia. Ninety-eight separate centrifugations of one to six hours duration were performed in 12 patients. Quantities of blood between 2.4-20.3 liters were processed during each centrifugation representing 0.4-3.2 donor blood volumes. An additional five centrifugations were attempted unsuccessfully. Buffy coat cells were collected while plasma and red blood cells were returned to the donor without change in flow or gravitational field. White blood cell yields of up to 75 per cent were obtained while platelet yields were electively varied from 0.1 per cent in donors with thrombocytopenia to 61.0 per cent in donors with thrombocytosis. Red blood cell hemolysis and excessive platelet losses with thrombocytopenia were not observed. One patient developed chills and fever of unknown etiology on two separate occasions, once during and once following CFC Leukocytes collected by this method and transfused into granulocytopenic recipients resulted in granulocyte increments of 2.0 x 103 per mm3 per 1011 granulocytes per m2 of body surface area. The persistence of cells with the Ph1 chromosome in the bone marrow of recipients demonstrated that viable proliferating cells survived the procedure.
A closed continuous flow centrifuge was used to separate and collect large quantities of buffy coat cells from the dog. One hundred fifty-five separate centrifugations of 2-12 hours duration were performed. Up to 61.0 liters of blood, representing 2.2-52 donor blood volumes, were processed. Buffy coat cells with a preponderance of granulocytes or lymphocytes, were collected while plasma and red blood cells were returned to the donor without change in flow or gravitational field. The mean total number of leukocytes, granulocytes, mononuclear cells and platelets removed was 24.0, 17.0, 7.0 and 197.0 x 10,9 respectively, which was 34.0, 28.0, 66.0 and 36.0 percent, respectively, of each cell population entering the centrifuge. The effect of centrifugation on blood components was evaluated. Granulocytes from buffy coat collections exhibited normal phagocytic ability in vitro. The transfusion of large quantities of granulocytes (15.0 x 109) into leukopenic dogs produced an increment in peripheral granulocyte count (2.7 x 103 per mm.3). Granulocytes labeled with 3H-DFP were also infused with a recovery of 38.6 percent at one hour and a T-½ of 4-6 hours. Machine-separated lymphocytes responded normally to phytohemagglutinin. Prolonged passage of blood through the pumps, tubing and bowl without centrifugation resulted in a decrease in circulating platelet levels (21 percent). A greater decrease in platelets occurred with centrifugation at high g. forces (49 percent). Hemolysis of red blood cells was not a serious problem.
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