Characteristics of lympho-myelopoietic stem cells isolated from canine peripheral blood.
If hematopoietic stem cells (HSC) could be separated from peripheral blood, it might be possible to harvest these stem cells for potential clinical use. By leukapheresis techniques, we harvested mononuclear cells (MNC) from peripheral blood and then placed these cells over discontinuous stractan gradients of three densities (1.077 gm/ml, 1.071 gm/ml and 1.066 gm/ml). These separated cells were submitted to colony culture to identify colony-forming-unit activity for granulocyte-macrophage (CFU-C) and T-cell lymphocyte (CFU-L) cell lines. The lightest cells (1.066) contained most of the CFU-C and no CFU-L activity. Heavier cells (greater than 1.071) contained CFU-L and very little CFU-C activity. CFU-L colonies could be distinguished from CFU-C by their density and distinct morphological appearance. In addition, the amount of CFU-C could be increased in the animal by increasing the amount of blood processed (from 3.9 +/- .76 CFU-C/10(6) MNC to 6.7 +/- .35 CFU-C/10(6) MNC). This resulted in an increase of CFU-C collected from 7.6 +/- 2.1 CFU-C/10(6) MNC after the first equivalent blood volume to 22.5 +/- 3.4 CFU-C/10(6) MNC after the third equivalent blood volume processed. These results suggest that leukapheresis and gradient density separation may be useful procedures to obtain HSC.
D000818:Animals; D001853:Bone Marrow / Q000502:physiology; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D004285:Dogs; D006412:Hematopoietic Stem Cells / Q000502:physiology* / Q000648:ultrastructure; D007937:Leukapheresis / Q000379:methods; D008214:Lymphocytes / Q000502:physiology*; D008854:Microscopy, Electron; D009000:Monocytes / Q000166:cytology