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Isolation and identification of two CD34+ cell subpopulations from normal human peripheral blood

Isolation and identification of two CD34+ cell subpopulations from normal human peripheral blood Abstract Circulating CD34+ progenitors were separated from normal human peripheral blood on the basis of size and density by counterflow centrifugal elutriation (CCE). The CD34+ cells, 0.15% of peripheral blood mononuclear cells, were heterogeneous with respect to their elutriation characteristics, mainly size and density. The least mature CD34+ cells, characterized by lack of CD38 antigen, were predominantly found in the small lymphoid cell fraction. In fractions containing larger and denser cells (large lymphocytes, monocytes, and granulocytes), CD38 was increasingly expressed on the CD34+ cells, as were lineage commitment markers CD10 (B lymphoid), CD33 (myeloid), CD13 (myelomonocytic) and CD71 (erythroid) antigens. The smaller and less dense CD34+ cells expressed CD34 antigen brightly while the larger and denser CD34+ cells expressed it dimly. The smaller and less dense CD34+high cells failed to establish colony growth in short-term culture while the larger and denser CD34+low cells gave rise to high counts of colony forming units-granulocyte macrophage (CFU-GM). Physical separation on the basis of size and density by CCE differentiates between two main classes of steadystate CD34+ cells from normal human peripheral blood. The smaller and less dense CD34+high cells correspond to the earliest progenitors that express differentiation markers poorly but CD34 antigen brightly, do not give rise to short-term colony growth in vitro, and thus represent indirect evidence for pluripotent hematopoietic stem cells (PHSC). The larger and denser CD34+low cells are the more mature progenitor cells, already committed to myeloid, lymphoid or erythroid differentiation but only dimly expressing CD34 antigen, and these cells were responsible for short-term colony growth in vitro. Normal peripheral blood, Hématopoietic progenitor cells, CD34 antigen, Pluripotent hematopoietic stem cells, Counterflow centrifugal elutriation References 1 Suda T , Suda J, Ogawa M. Single-cell origin of mouse hemopoietic colonies expressing multiple lineages in variable combinations . Proc Natl Acad Sci USA 1983 ; 80 : 6689 – 6693 . Google Scholar Crossref Search ADS PubMed WorldCat 2 Spangrude GJ , Heimfeld S, Weissman IL. Purification and characterization of mouse hematopoietic stem cells . Science 1988 ; 241 : 58 – 62 . Google Scholar Crossref Search ADS PubMed WorldCat 3 Harrison DE . Competitive repopulation: A new assay for long-term stem cell functional capacity . Blood 1980 ; 55 : 77 – 81 . Google Scholar Crossref Search ADS PubMed WorldCat 4 Harrison D , Astle CM. Lymphoid and erythroid repopulation in B6 W-anemic mice: A new unirradiated recipient . Exp Hematol 1991 ; 19 : 374 – 377 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 5 Barker JE , Greer J, Bacon S, Compton ST. Temporal replacement of donor erythrocytes and leukocytes in nonanemic W44j/W44j and severely anemic W/Wv mice . Blood 1991 ; 78 : 1432 – 1437 . Google Scholar Crossref Search ADS PubMed WorldCat 6 Jones RJ , Wagner JE, Celano P, Zicha MS, Sharkis SJ. Separation of pluripotent haematopoietic stem cells from spleen colony-forming cells . Nature 1990 ; 347 : 188 – 189 . Google Scholar Crossref Search ADS PubMed WorldCat 7 Hénon P , Liang H, Beck-Wirth G, Eisenmann JC, Lepers M, Wunder E, Kandel G. Comparison of hematopoietic and immune recovery after autologous bone marrow or blood stem cell transplants . Bone Marrow Transplant 1992 ; 9 : 285 – 291 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 8 Sovalat H , Liang H, Wunder E, Hénon P. Flow cytometry characterization of CD34 positive cells in bone marrow, cytapheresis products and cord blood at birth . Int J Cell Cloning 1992 ; 10 ( 1 ): 20 – 22 . Google Scholar OpenURL Placeholder Text WorldCat 9 McCarthy DM , Goldman JM. Transfusion of circulating stem cells . CRC Crit Rev Clin Lab Sci 1984 ; 20 : 1 – 24 . Google Scholar Crossref Search ADS WorldCat 10 Micklem HS , Anderson N, Ross E. Limited potential of circulating haematopoietic stem cells . Nature 1975 ; 256 : 41 – 43 . Google Scholar Crossref Search ADS PubMed WorldCat 11 To LB , Haylock DN, Kimber RJ, Juttner CA. High levels of circulating haematopoietic stem cells in very early remission from acute non-lymphoblastic leukaemia and their collection and cryopreservation . Br J Haematol 1984 ; 58 : 399 – 410 . Google Scholar Crossref Search ADS PubMed WorldCat 12 To LB , Dyson PG, Juttner CA. Cell dose effect in circulating stem cell autografting . Lancet 1986 ; 2 : 404 – 405 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 13 Civin CI , Banquerigo ML, Strauss LC, Loken MR. Antigenic analysis of hematopoiesis. VI. Flow cytometric characterization of MY 10 positive progenitor cells in normal bone marrow . Exp Hematol 1987 ; 15 : 10 – 17 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 14 Bender JG , Unverzagt KL, Walker DE, Lee W, Van Epp DE, Smith DH, Stewart CC, To LB. Identification and comparison of CD34 positive cells and their subpopulations from normal peripheral blood and bone marrow using multicolor flow cytometry . Blood 1991 ; 77 : 2591 – 2596 . Google Scholar Crossref Search ADS PubMed WorldCat 15 Sparrow RL , Williams N. The pattern of HLA-DR and HLA-DQ antigen expression on clonable subpopulations of human myeloid progenitor cells . Blood 1986 ; 67 : 379 – 384 . Google Scholar Crossref Search ADS PubMed WorldCat 16 Lu L , Walker D, Broxmeyer HE, Hoffman R, Hu W, Walker E. Characterization of adult human marrow hematopoietic progenitors highly enriched by two-color cell sorting with MY10 and major histocompatibility class II monoclonal antibodies . J Immunol 1987 ; 139 : 1823 – 1829 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 17 Andrews RG , Singer JW, Bernstein ID. Precursors of colony forming cells in humans can be distinguished from colony forming cells by expression of the CD33 and CD34 antigens and light scatter properties . J Exp Med 1989 ; 169 : 1721 – 1731 . Google Scholar Crossref Search ADS PubMed WorldCat 18 Knapp W . Myeloid antigen report. In: Knapp W, Dorken B, and Gilks WR, eds. Leucocyte Typing IV. White cell differentiation antigens . Oxford : Oxford University Press , 1990 : 747 – 780 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 19 Wahl LM , Katona IM, Wilder RL, Winter CC, Haraoui B, Scher I, Wahl SM. Isolation of human mononuclear cell subsets by counterflow centrifugal elutriation (CCE). I. Characterization of B lymphocyte-, T lymphocyte- and monocyte-enriched fractions by flow cytometric analysis . Cell Immunol 1984 ; 85 : 373 – 383 . Google Scholar Crossref Search ADS PubMed WorldCat 20 Wunder E , Sovalat H, Liang H, Hénon P. The role of monocytes in the stimulation of progenitor cells. In: Dicke KA, Armitage JO, Dicke-Evinger MJ, eds. Autologous Bone Marrow Transplantation . University of Nebraska Medical Center 1991 : 881 – 892 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 21 Terstappen WMM , Huang S, Safford M, Lansdorp PM, Loken MR. Sequential generations of hematopoietic colonies derived from single non-lineage-committed CD34+CD38- progenitor cells . Blood 1991 ; 77 : 1218 – 1227 . Google Scholar Crossref Search ADS PubMed WorldCat 22 Böhring HJ , Asenbauer B, Katrilaka K, Hummel G, Busch FW. Sequential expression of CD34 and CD33 antigens on myeloid colony forming cells . Eur J Haematol 1989 ; 42 : 143 – 149 . Google Scholar Crossref Search ADS PubMed WorldCat 23 Sutherland JH , Eaves CJ, Eaves AC, Dragowska W, Lansdorp PM. Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesis in vitro . Blood 1989 ; 74 : 1563 – 1570 . Google Scholar Crossref Search ADS PubMed WorldCat 24 Wunder E , Sovalat H, Lepers M, Hénon P. The role of monocytes/macrophages in blood stem cell maturation: Studies with highly purified precursor (CD34+) cells . Bone Marrow Transplant 1990 ; 5 : 11 – 12 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 25 Wunder E , Thing-Mortensen B, Sovalat H, Hénon P, Chabord P. Role of mononuclear macrophages in the regulation of hematopoiesis: In vitro experiments and observations in vivo. In: Van Furth, ed. Hematopoietic growth factors and mononuclear phagocytes . Basel , Karger : 1993 : 36 – 43 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 26 Stamenkovic I , Seed B. CD19, the earliest differentiation antigen of the B cell lineage, bears three extracellular immunoglobulin like domains and an Epstein-Barr virus-related cytoplasmic tail . J Exp Med 1988 ; 168 : 1205 – 1210 . Google Scholar Crossref Search ADS PubMed WorldCat 27 Sieff C , Bicknell D, Caine G, Robinson J, Lam G, Greaves MF. Changes in cell surface antigen expression during hematopoietic differentiation . Blood 1982 ; 60 : 703 – 713 . Google Scholar Crossref Search ADS PubMed WorldCat 28 Loken MR , Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow. I. Normal erythroid development . Blood 1987 ; 69 : 255 – 263 . Google Scholar Crossref Search ADS PubMed WorldCat 29 Neckers LM , Cossman J. Transferrin receptor induction in mitogen stimulated human T lymphocytes is required for DNA synthesis and cell division and is regulated by interleukin 2 . Proc Natl Acad Sci USA 1983 ; 80 : 3494 – 3498 . Google Scholar Crossref Search ADS PubMed WorldCat This content is only available as a PDF. © 1994 AlphaMed Press This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Stem Cells Oxford University Press

Isolation and identification of two CD34+ cell subpopulations from normal human peripheral blood

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Oxford University Press
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Copyright © 2022 Oxford University Press
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1066-5099
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1549-4918
DOI
10.1002/stem.5530120207
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Abstract

Abstract Circulating CD34+ progenitors were separated from normal human peripheral blood on the basis of size and density by counterflow centrifugal elutriation (CCE). The CD34+ cells, 0.15% of peripheral blood mononuclear cells, were heterogeneous with respect to their elutriation characteristics, mainly size and density. The least mature CD34+ cells, characterized by lack of CD38 antigen, were predominantly found in the small lymphoid cell fraction. In fractions containing larger and denser cells (large lymphocytes, monocytes, and granulocytes), CD38 was increasingly expressed on the CD34+ cells, as were lineage commitment markers CD10 (B lymphoid), CD33 (myeloid), CD13 (myelomonocytic) and CD71 (erythroid) antigens. The smaller and less dense CD34+ cells expressed CD34 antigen brightly while the larger and denser CD34+ cells expressed it dimly. The smaller and less dense CD34+high cells failed to establish colony growth in short-term culture while the larger and denser CD34+low cells gave rise to high counts of colony forming units-granulocyte macrophage (CFU-GM). Physical separation on the basis of size and density by CCE differentiates between two main classes of steadystate CD34+ cells from normal human peripheral blood. The smaller and less dense CD34+high cells correspond to the earliest progenitors that express differentiation markers poorly but CD34 antigen brightly, do not give rise to short-term colony growth in vitro, and thus represent indirect evidence for pluripotent hematopoietic stem cells (PHSC). The larger and denser CD34+low cells are the more mature progenitor cells, already committed to myeloid, lymphoid or erythroid differentiation but only dimly expressing CD34 antigen, and these cells were responsible for short-term colony growth in vitro. Normal peripheral blood, Hématopoietic progenitor cells, CD34 antigen, Pluripotent hematopoietic stem cells, Counterflow centrifugal elutriation References 1 Suda T , Suda J, Ogawa M. Single-cell origin of mouse hemopoietic colonies expressing multiple lineages in variable combinations . Proc Natl Acad Sci USA 1983 ; 80 : 6689 – 6693 . Google Scholar Crossref Search ADS PubMed WorldCat 2 Spangrude GJ , Heimfeld S, Weissman IL. Purification and characterization of mouse hematopoietic stem cells . Science 1988 ; 241 : 58 – 62 . Google Scholar Crossref Search ADS PubMed WorldCat 3 Harrison DE . Competitive repopulation: A new assay for long-term stem cell functional capacity . Blood 1980 ; 55 : 77 – 81 . Google Scholar Crossref Search ADS PubMed WorldCat 4 Harrison D , Astle CM. Lymphoid and erythroid repopulation in B6 W-anemic mice: A new unirradiated recipient . Exp Hematol 1991 ; 19 : 374 – 377 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 5 Barker JE , Greer J, Bacon S, Compton ST. Temporal replacement of donor erythrocytes and leukocytes in nonanemic W44j/W44j and severely anemic W/Wv mice . Blood 1991 ; 78 : 1432 – 1437 . Google Scholar Crossref Search ADS PubMed WorldCat 6 Jones RJ , Wagner JE, Celano P, Zicha MS, Sharkis SJ. Separation of pluripotent haematopoietic stem cells from spleen colony-forming cells . Nature 1990 ; 347 : 188 – 189 . Google Scholar Crossref Search ADS PubMed WorldCat 7 Hénon P , Liang H, Beck-Wirth G, Eisenmann JC, Lepers M, Wunder E, Kandel G. Comparison of hematopoietic and immune recovery after autologous bone marrow or blood stem cell transplants . Bone Marrow Transplant 1992 ; 9 : 285 – 291 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 8 Sovalat H , Liang H, Wunder E, Hénon P. Flow cytometry characterization of CD34 positive cells in bone marrow, cytapheresis products and cord blood at birth . Int J Cell Cloning 1992 ; 10 ( 1 ): 20 – 22 . Google Scholar OpenURL Placeholder Text WorldCat 9 McCarthy DM , Goldman JM. Transfusion of circulating stem cells . CRC Crit Rev Clin Lab Sci 1984 ; 20 : 1 – 24 . Google Scholar Crossref Search ADS WorldCat 10 Micklem HS , Anderson N, Ross E. Limited potential of circulating haematopoietic stem cells . Nature 1975 ; 256 : 41 – 43 . Google Scholar Crossref Search ADS PubMed WorldCat 11 To LB , Haylock DN, Kimber RJ, Juttner CA. High levels of circulating haematopoietic stem cells in very early remission from acute non-lymphoblastic leukaemia and their collection and cryopreservation . Br J Haematol 1984 ; 58 : 399 – 410 . Google Scholar Crossref Search ADS PubMed WorldCat 12 To LB , Dyson PG, Juttner CA. Cell dose effect in circulating stem cell autografting . Lancet 1986 ; 2 : 404 – 405 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 13 Civin CI , Banquerigo ML, Strauss LC, Loken MR. Antigenic analysis of hematopoiesis. VI. Flow cytometric characterization of MY 10 positive progenitor cells in normal bone marrow . Exp Hematol 1987 ; 15 : 10 – 17 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 14 Bender JG , Unverzagt KL, Walker DE, Lee W, Van Epp DE, Smith DH, Stewart CC, To LB. Identification and comparison of CD34 positive cells and their subpopulations from normal peripheral blood and bone marrow using multicolor flow cytometry . Blood 1991 ; 77 : 2591 – 2596 . Google Scholar Crossref Search ADS PubMed WorldCat 15 Sparrow RL , Williams N. The pattern of HLA-DR and HLA-DQ antigen expression on clonable subpopulations of human myeloid progenitor cells . Blood 1986 ; 67 : 379 – 384 . Google Scholar Crossref Search ADS PubMed WorldCat 16 Lu L , Walker D, Broxmeyer HE, Hoffman R, Hu W, Walker E. Characterization of adult human marrow hematopoietic progenitors highly enriched by two-color cell sorting with MY10 and major histocompatibility class II monoclonal antibodies . J Immunol 1987 ; 139 : 1823 – 1829 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 17 Andrews RG , Singer JW, Bernstein ID. Precursors of colony forming cells in humans can be distinguished from colony forming cells by expression of the CD33 and CD34 antigens and light scatter properties . J Exp Med 1989 ; 169 : 1721 – 1731 . Google Scholar Crossref Search ADS PubMed WorldCat 18 Knapp W . Myeloid antigen report. In: Knapp W, Dorken B, and Gilks WR, eds. Leucocyte Typing IV. White cell differentiation antigens . Oxford : Oxford University Press , 1990 : 747 – 780 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 19 Wahl LM , Katona IM, Wilder RL, Winter CC, Haraoui B, Scher I, Wahl SM. Isolation of human mononuclear cell subsets by counterflow centrifugal elutriation (CCE). I. Characterization of B lymphocyte-, T lymphocyte- and monocyte-enriched fractions by flow cytometric analysis . Cell Immunol 1984 ; 85 : 373 – 383 . Google Scholar Crossref Search ADS PubMed WorldCat 20 Wunder E , Sovalat H, Liang H, Hénon P. The role of monocytes in the stimulation of progenitor cells. In: Dicke KA, Armitage JO, Dicke-Evinger MJ, eds. Autologous Bone Marrow Transplantation . University of Nebraska Medical Center 1991 : 881 – 892 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 21 Terstappen WMM , Huang S, Safford M, Lansdorp PM, Loken MR. Sequential generations of hematopoietic colonies derived from single non-lineage-committed CD34+CD38- progenitor cells . Blood 1991 ; 77 : 1218 – 1227 . Google Scholar Crossref Search ADS PubMed WorldCat 22 Böhring HJ , Asenbauer B, Katrilaka K, Hummel G, Busch FW. Sequential expression of CD34 and CD33 antigens on myeloid colony forming cells . Eur J Haematol 1989 ; 42 : 143 – 149 . Google Scholar Crossref Search ADS PubMed WorldCat 23 Sutherland JH , Eaves CJ, Eaves AC, Dragowska W, Lansdorp PM. Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesis in vitro . Blood 1989 ; 74 : 1563 – 1570 . Google Scholar Crossref Search ADS PubMed WorldCat 24 Wunder E , Sovalat H, Lepers M, Hénon P. The role of monocytes/macrophages in blood stem cell maturation: Studies with highly purified precursor (CD34+) cells . Bone Marrow Transplant 1990 ; 5 : 11 – 12 . Google Scholar PubMed OpenURL Placeholder Text WorldCat 25 Wunder E , Thing-Mortensen B, Sovalat H, Hénon P, Chabord P. Role of mononuclear macrophages in the regulation of hematopoiesis: In vitro experiments and observations in vivo. In: Van Furth, ed. Hematopoietic growth factors and mononuclear phagocytes . Basel , Karger : 1993 : 36 – 43 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC 26 Stamenkovic I , Seed B. CD19, the earliest differentiation antigen of the B cell lineage, bears three extracellular immunoglobulin like domains and an Epstein-Barr virus-related cytoplasmic tail . J Exp Med 1988 ; 168 : 1205 – 1210 . Google Scholar Crossref Search ADS PubMed WorldCat 27 Sieff C , Bicknell D, Caine G, Robinson J, Lam G, Greaves MF. Changes in cell surface antigen expression during hematopoietic differentiation . Blood 1982 ; 60 : 703 – 713 . Google Scholar Crossref Search ADS PubMed WorldCat 28 Loken MR , Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow. I. Normal erythroid development . Blood 1987 ; 69 : 255 – 263 . Google Scholar Crossref Search ADS PubMed WorldCat 29 Neckers LM , Cossman J. Transferrin receptor induction in mitogen stimulated human T lymphocytes is required for DNA synthesis and cell division and is regulated by interleukin 2 . Proc Natl Acad Sci USA 1983 ; 80 : 3494 – 3498 . Google Scholar Crossref Search ADS PubMed WorldCat This content is only available as a PDF. © 1994 AlphaMed Press This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

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Stem Cells – Oxford University Press

Published: Jan 1, 1994

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