Blood Cell Separation in the Dog by Continuous Flow Centrifugation

Abstract

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.

Collection of Peripheral Blood CD34+ Progenitor Cells from Healthy Dogs and Dogs Diagnosed with Lymphoproliferative Diseases Using a Baxter‐Fenwal CS‐3000 Plus Blood Cell Separator

“…[7][8][9] Development of techniques to both increase peripheral blood mononuclear cell (PBMC) counts using cloned hematopoietic cytokines 10,11 and collect large numbers of these cells via centrifugation using continuous-flow blood separators increased yields. [12][13][14] In dogs, both autologous 15,16 and allogeneic 17 PBMC grafts were able to reconstitute normal hematopoiesis after lethal bone marrow ablation. Freshly isolated PBMC grafts were used to treat dogs in remission for LSA after high-dose total body irradiation, with approximately 25% becoming disease-free long-term survivors.…”

Principles of Peripheral Blood Mononuclear Cell Apheresis in a Preclinical Canine Model of Hematopoietic Cell Transplantation

“…F or more than 5 decades, the dog has served as an excellent random-bred preclinical model for human hematopoietic cell transplantation (HCT). 1,2 Studies in the canine model concerning the impact of histocompatibility barriers on engraftment and graft-versus-host disease (GVHD), 3 in vivo [4][5][6][7][8] and in vitro [9][10][11][12] graft manipulation, peripheral blood mononuclear cell (PBMC) collection, [13][14][15][16][17][18][19] kinetics of immunological reconstitution after bone marrow and PBMC grafting, 20,21 cell dose requirements, [21][22][23][24] efficacy of various pretransplant conditioning regimens [25][26][27][28][29][30][31][32][33][34] and posttransplant immunosuppression protocols, [35][36][37][38][39] treatment of malignant diseases, [40][41][42][43][44][45][46][47][48] and graft-versus-tumor (GVT...…”

“…15 Since then, various PBMC collection and isolation methods have been adopted for the canine HCT model. 2,11,12,14,[16][17][18][19] Extensive preclinical studies showed that both autologous 10,20,21,53 and allogeneic 3,13 PBMC grafts reconstituted normal hematopoiesis in dogs undergoing otherwise lethal bone marrow ablation. It was also established that 10 times more PBMCs than marrow cells were required to induce regeneration of hematopoietic cell lineages in a timely fashion and assure stable engraftment.…”

The effect of lymphocyte depletion by continuous flow centrifugation in canine renal allotransplants

“…The NCI-IBM continuous flow blood cell centrifuge was adapted for use in the dog, based in part on the recent findings of Judson, Jones, Buckner, and associates (Judson et al, 1968;Jones, 1968;Buckner et al, 1968).…”