Abstract
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 (>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/106 MNC to 6.7±.35 CFU-C/106 MNC). This resulted in an increase of CFU-C collected from 7.6±2.1 CFU-C/106 MNC after the first equivalent blood volume to 22.5±3.4 CFU-C/106 MNC after the third equivalent blood volume processed. These results suggest that leukapheresis and gradient density separation may be useful procedures to obtain HSC.
To whom correspondence should be addressed
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Abbreviations
- (HSC):
-
Hematopoietic Stem Cells
- (MNC):
-
Mononuclear Cells
- (CFU-L):
-
Colony-Forming-Units for Lymphocytes
- (CFU-C):
-
granulocytes
- (GvH):
-
Graft versus Host Disease
References
E.D. Thomas, R. Storb, R.A. Clift, A. Feder, L. Johnson, P.E. Neiman, K.G. Lerner, H. Glucksberg and C.D. Buckner, Bone-marrow transplantation (first of two parts). N. Engl. Med. 292:832–843, 1975.
E.D. Thomas, R. Storb, R.A. Clift, A. Feder, L. Johnson, P.E. Neiman, K.G. Lerner, H. Glucksberg and C.D. Buckner, Bone-marrow transplantation (second of two parts). N. Engl. Med. 292:895–902, 1975.
R.P. Gale, R.E. Champlin, S.A. Feig and J.H. Fitchen, Aplastic anemia: biology and treatment. Ann. Intern Med. 95:477–494, 1981.
S.C. Grebe and J.W. Streilan, Graft-versus-host reactions: a review. Adv. Immunol. 22.19–221, 1976.
American Association of Blood Banks, Technical Manual. Chapter 2, 1977.
L. Corash, H. Tan and H.R. Gralnick, Heterogeneity of human whole blood platelet subpopulations. I. Relationship between buoyant density, cell volume, and ultrastructure. Blood 49:71–77, 1977.
M. Porvaznik and T.J. MacVittie, Detection of gap junctions between the progeny of canine macrophage colony-forming cell in vitro. J. Cell. Biol. 82:555–564, 1979.
F.D. Wilson, J.A. Dyck, S.J. Knox and M. Shifrine, A “whole blood” technique for the quantitation of canine “T-lymphocyte” progenitors using a semi-solid culture system. Exp. Hematol. 8:1031–1039, 1980.
T.J. MacVittie and R.I. Walker, Endotoxin-induced alterations in canine granulopoiesis colony stimulating factor, colony-forming cells in culture, and growth of cells in diffusion chamber, Exp. Hematol. 6:613–618, 1978.
R.A. Abrams and A.M. Deisseroth, Prospects for accelerating hematopoietic recovery following myelosuppressive therapy by using autologous, cyropreserved hematopoietic stem cells collected solely from the peripheral blood. Exp. Hematol. 7:(supp. 5): 1107–1115, 1979.
T.M. Fliedner, W. Calvo, M. Körbling, W. Nothdurft, H. Pflieger and W. Ross, Collection, storage and transfusion of blood stem cells for the treatment of hematopoietic failure. Blood Cells 5: 313–328, 1979.
R.S. Weiner, C.M. Richman and R.A. Yankee, Semicontinuous flow centrifugation for the pheresis of immunocompetent cells and stem cells. Blood 49:391–397, 1977.
S.C. Sarpel, A.R. Zander, L. Horvath and R.B. Epstein, The collection preservation, and function of peripheral blood hematopoietic cells in dogs. Exp. Hematol. 7:113–120, 1979.
R.A. Abrams, D. Glaubiger, F.R. Appelbaum and A.B. Deisseroth, Result of attempted hematopoietic reconstitution using isologous, peripheral blood mononuclear cells: a case report. Blood 56:516–520, 1980.
C.R. Richman, R.S. Weiner and R.A. Yankee, Increase in circulating stem cells following chemotherapy in man. Blood 47:1031–1039, 1976.
E.D. Thomas and R. Storb, Technique for human marrow grafting. Blood 36:507–515, 1970.
K.A. Dicke and D.W. van Bekkum, Allogeneic bone marrow transplantation after elimination of immunocompetent cells by means of density gradient centrifugation. Transplant Proceedings, III:666–668, 1971.
R.A. Phillips and R.G. Miller, Physical separation of hematopoietic stem cells from cells causing graft-vs-host disease. I. Sedimentation properties of cells causing graft-vs-host disease. J. Immunol. 105:1168–1174, 1970.
J.R. Wells, G. Opelz and M.J. Cline, Characterization of functionally distinct lymphoid and myeloid cells from human blood and bone marrow. I. Separation by a buoyant density gradient technique. J. Immunol. Methods 18:63–77, 1977.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1984 Plenum Press, New York
About this chapter
Cite this chapter
Stewart, D.A., Bolin, R.B., Cheney, B.A., Hawkins, J.T., Chapman, K.W., Tompkins, D.R. (1984). Characteristics of Lympho-Myelopoietic Stem Cells Isolated from Canine Peripheral Blood. In: Acton, R.T., Daniel Lynn, J. (eds) Eukaryotic Cell Cultures. Advances in Experimental Medicine and Biology, vol 172. Springer, New York, NY. https://doi.org/10.1007/978-1-4615-9376-8_3
Download citation
DOI: https://doi.org/10.1007/978-1-4615-9376-8_3
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4615-9378-2
Online ISBN: 978-1-4615-9376-8
eBook Packages: Springer Book Archive