The collection, preservation and function of peripheral blood hematopoietic cells in dogs
Sarpel, S.C.; Zander, A.R.; Harvath, L.; Epstein, R.B.
Experimental Hematology 7(2): 113-120
1979
ISSN/ISBN: 0301-472X PMID: 371973 Document Number: 152321
Semicontinuous flow centrifugation (SFC) was employed in canines to obtain adequate numbers of cells for autologous marrow repopulation following supralethal cyclophosphamide administration (100 mg/kg). Four cycles using a 225 ml bowl and 30 ml/min flow rate were carried out for procurement. Collections averaged 8.4 .+-. 0.4 .times. 109 (n = 30) leukocytes. Mononuclear cells (MNC) comprised 75 .+-. 2% of the population and granulocyte colony-forming units (CFU-C) totaled 1.9 .+-. 0.09 .times. 105 colonies per collection. Of mononuclear cells, 86% were T [thymus-derived] cells in the 30-90 s fraction compared to 49% at 150-120 s. CFU-C fractionation revealed a peak at 30-210 s and a 2nd peak at 120-210 s. Following programmed freezing and rapid thawing 77.7 .+-. 11.4% of CFU-C were recovered. Using a dose of 1 .times. 109 MNC/kg for reinfusion, marrow repopulation and clinical recovery occurred in 4 out of 4 dogs. SFC was effective for obtaining adequate numbers of peripheral blood stem cells for autologous marrow repopulation. A rapid thawing and direct transfusion technique appears satisfactory for administration. Some separation of T, B [bone marrow-derived] and CFU-C peripheral blood components is possible by centrifugation. [The transplantation of allogeneic hematopoietic tissue for treatment of hematologic diseases (aplastic anemia, immunodeficiency syndrome, acute leukemia) was discussed.].