Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1989 May 1;169(5):1721–1731. doi: 10.1084/jem.169.5.1721

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

PMCID: PMC2189320  PMID: 2469766

Abstract

We determined whether human marrow cells that directly form colonies in vitro could be distinguished from cells that generate or become CFC only after LTMC in the presence of irradiated marrow stromal cells. In previous studies, an anti-CD33 antibody, L4F3, and complement (C') were used to lyse nearly all CFC in marrow, and the remaining cells generated CFC in LTMC. In the present studies, marrow cells were treated with L4F3 + C' and the remaining CD33- cells were separated into CD34+ and CD34- populations and placed in LTMC. Only the CD34+ cells were found to generate significant numbers of CFC. To compare the CD33-CD34+ and CD33+CD34+ cells, we isolated each cell population using two-color FACS. Only LTMCs of the CD33-CD34+ cells generated CFC for greater than 5 wk. In contrast, cells that expressed both the CD33 and CD34 antigens, which contained most of the CFC, generated few CFC in LTMC. Fractionation of marrow cells based on right angle and forward light scattering suggested that precursors for CFC have low right angle and low forward light scattering properties. The CD33-CD34+ marrow cells were therefore further fractionated based on light scatter characteristics. Cells with low right angle and low forward light scatter formed few or no colonies on direct culture, yet generated greater numbers of CFC after 4 wk of LTMC than did cells with low right angle and high forward light scatter. Most (87-98%) CFC generated in the LTMCs that were initiated with CD33-CD34+ cells were found to express the CD33 antigen. Thus, hematopoietic progenitors with differing proliferative and differentiative potentials can be directly separated on the basis of their expression of CD33 and CD34 cell surface antigens and their light scatter properties.

Full Text

The Full Text of this article is available as a PDF (788.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews R. G., Singer J. W., Bernstein I. D. Monoclonal antibody 12-8 recognizes a 115-kd molecule present on both unipotent and multipotent hematopoietic colony-forming cells and their precursors. Blood. 1986 Mar;67(3):842–845. [PubMed] [Google Scholar]
  2. Andrews R. G., Takahashi M., Segal G. M., Powell J. S., Bernstein I. D., Singer J. W. The L4F3 antigen is expressed by unipotent and multipotent colony-forming cells but not by their precursors. Blood. 1986 Nov;68(5):1030–1035. [PubMed] [Google Scholar]
  3. Andrews R. G., Torok-Storb B., Bernstein I. D. Myeloid-associated differentiation antigens on stem cells and their progeny identified by monoclonal antibodies. Blood. 1983 Jul;62(1):124–132. [PubMed] [Google Scholar]
  4. Berenson R. J., Andrews R. G., Bensinger W. I., Kalamasz D., Knitter G., Buckner C. D., Bernstein I. D. Antigen CD34+ marrow cells engraft lethally irradiated baboons. J Clin Invest. 1988 Mar;81(3):951–955. doi: 10.1172/JCI113409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bodger M. P., Izaguirre C. A., Blacklock H. A., Hoffbrand A. V. Surface antigenic determinants on human pluripotent and unipotent hematopoietic progenitor cells. Blood. 1983 May;61(5):1006–1010. [PubMed] [Google Scholar]
  6. Civin C. I., Strauss L. C., Brovall C., Fackler M. J., Schwartz J. F., Shaper J. H. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells. J Immunol. 1984 Jul;133(1):157–165. [PubMed] [Google Scholar]
  7. Denis K. A., Witte O. N. In vitro development of B lymphocytes from long-term cultured precursor cells. Proc Natl Acad Sci U S A. 1986 Jan;83(2):441–445. doi: 10.1073/pnas.83.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Denkers E. Y., Badger C. C., Ledbetter J. A., Bernstein I. D. Influence of antibody isotype on passive serotherapy of lymphoma. J Immunol. 1985 Sep;135(3):2183–2186. [PubMed] [Google Scholar]
  9. Dick J. E., Magli M. C., Huszar D., Phillips R. A., Bernstein A. Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/Wv mice. Cell. 1985 Aug;42(1):71–79. doi: 10.1016/s0092-8674(85)80102-1. [DOI] [PubMed] [Google Scholar]
  10. Ferrero D., Broxmeyer H. E., Pagliardi G. L., Venuta S., Lange B., Pessano S., Rovera G. Antigenically distinct subpopulations of myeloid progenitor cells (CFU-GM) in human peripheral blood and marrow. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4114–4118. doi: 10.1073/pnas.80.13.4114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ferrero D., Gabbianelli M., Peschle C., Lange B., Rovera G. Surface phenotypes of human hemopoietic progenitor cells defined by monoclonal antibodies. Blood. 1985 Sep;66(3):496–502. [PubMed] [Google Scholar]
  12. Griffin J. D., Linch D., Sabbath K., Larcom P., Schlossman S. F. A monoclonal antibody reactive with normal and leukemic human myeloid progenitor cells. Leuk Res. 1984;8(4):521–534. doi: 10.1016/0145-2126(84)90001-8. [DOI] [PubMed] [Google Scholar]
  13. Griffin J. D., Ritz J., Nadler L. M., Schlossman S. F. Expression of myeloid differentiation antigens on normal and malignant myeloid cells. J Clin Invest. 1981 Oct;68(4):932–941. doi: 10.1172/JCI110348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson A., Dorshkind K. Stromal cells in myeloid and lymphoid long-term bone marrow cultures can support multiple hemopoietic lineages and modulate their production of hemopoietic growth factors. Blood. 1986 Dec;68(6):1348–1354. [PubMed] [Google Scholar]
  15. Katz F. E., Tindle R., Sutherland D. R., Greaves M. F. Identification of a membrane glycoprotein associated with haemopoietic progenitor cells. Leuk Res. 1985;9(2):191–198. doi: 10.1016/0145-2126(85)90082-7. [DOI] [PubMed] [Google Scholar]
  16. Loken M. R., Shah V. O., Dattilio K. L., Civin C. I. Flow cytometric analysis of human bone marrow. II. Normal B lymphocyte development. Blood. 1987 Nov;70(5):1316–1324. [PubMed] [Google Scholar]
  17. Look A. T., Peiper S. C., Douglass E. C., Trent J. M., Sherr C. J. Amplification of genes encoding human myeloid membrane antigens after DNA-mediated gene transfer. Blood. 1986 Mar;67(3):637–645. [PubMed] [Google Scholar]
  18. McKearn J. P., McCubrey J., Fagg B. Enrichment of hematopoietic precursor cells and cloning of multipotential B-lymphocyte precursors. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7414–7418. doi: 10.1073/pnas.82.21.7414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Papayannopoulou T., Brice M., Yokochi T., Rabinovitch P. S., Lindsley D., Stamatoyannopoulos G. Anti-HEL cell monoclonal antibodies recognize determinants that are also present in hemopoietic progenitors. Blood. 1984 Feb;63(2):326–334. [PubMed] [Google Scholar]
  20. Powell J. S., Fialkow P. J., Adamson J. W. Human mixed cell colonies: unicellular or multicellular origin--analysis by G-6-PD. Br J Haematol. 1984 May;57(1):89–95. [PubMed] [Google Scholar]
  21. Ryan D. H., Chapple C. W., Kossover S. A., Sandberg A. A., Cohen H. J. Phenotypic similarities and differences between CALLA-positive acute lymphoblastic leukemia cells and normal marrow CALLA-positive B cell precursors. Blood. 1987 Sep;70(3):814–821. [PubMed] [Google Scholar]
  22. Schlunk T., Schleyer M. The influence of culture conditions on the production of colony-stimulating activity by human placenta. Exp Hematol. 1980 Feb;8(2):179–184. [PubMed] [Google Scholar]
  23. Strauss L. C., Brovall C., Fackler M. J., Schwartz J. F., Shaper J. H., Loken M. R., Civin C. I. Antigenic analysis of hematopoiesis. IV. The My-11 hematopoietic cell surface antigen is expressed by myelomonocytic and lymphoid, but not erythroid, progenitor cells. Exp Hematol. 1986 Nov;14(10):935–945. [PubMed] [Google Scholar]
  24. Strauss L. C., Rowley S. D., La Russa V. F., Sharkis S. J., Stuart R. K., Civin C. I. Antigenic analysis of hematopoiesis. V. Characterization of My-10 antigen expression by normal lymphohematopoietic progenitor cells. Exp Hematol. 1986 Oct;14(9):878–886. [PubMed] [Google Scholar]
  25. Strauss L. C., Stuart R. K., Civin C. I. Antigenic analysis of hematopoiesis. I. Expression of the My-1 granulocyte surface antigen on human marrow cells and leukemic cell lines. Blood. 1983 Jun;61(6):1222–1231. [PubMed] [Google Scholar]
  26. TILL J. E., McCULLOCH E. A. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat Res. 1961 Feb;14:213–222. [PubMed] [Google Scholar]
  27. Tindle R. W., Nichols R. A., Chan L., Campana D., Catovsky D., Birnie G. D. A novel monoclonal antibody BI-3C5 recognises myeloblasts and non-B non-T lymphoblasts in acute leukaemias and CGL blast crises, and reacts with immature cells in normal bone marrow. Leuk Res. 1985;9(1):1–9. doi: 10.1016/0145-2126(85)90016-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES