Human Primitive Hematopoietic Progenitor Cells Are More Enriched in KITlow Cells Than in KIThigh Cells

To clarify the phenotypes of various classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with CD34 and a newly developed mouse antihuman c-kit proto-oncogene product (KIT) monoclonal antibody (MoAb). We characterized three cell fractions in CD34+ ce...

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Bibliographic Details
Published in:Blood 1993-12, Vol.82 (11), p.3283-3289
Main Authors: Gunji, Yuji, Nakamura, Mitsuru, Osawa, Hisao, Nagayoshi, Kazunari, Nakauchi, Hiromitsu, Miura, Yasusada, Yanagisawa, Masayoshi, Suda, Toshio
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - immunology
Antigens, CD - analysis
Antigens, CD34
Biological and medical sciences
Cell differentiation, maturation, development, hematopoiesis
Cell physiology
Cell Separation
Cells, Cultured
Fundamental and applied biological sciences. Psychology
Hematopoiesis
Hematopoietic Stem Cells - chemistry
Humans
Mice
Molecular and cellular biology
Proto-Oncogene Proteins - analysis
Proto-Oncogene Proteins - immunology
Proto-Oncogene Proteins c-kit
Receptor Protein-Tyrosine Kinases - analysis
Receptor Protein-Tyrosine Kinases - immunology
Receptors, Colony-Stimulating Factor - analysis
Receptors, Colony-Stimulating Factor - immunology
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Summary:To clarify the phenotypes of various classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with CD34 and a newly developed mouse antihuman c-kit proto-oncogene product (KIT) monoclonal antibody (MoAb). We characterized three cell fractions in CD34+ cells that express KITlow and KIThigh cells in addition to KIT– cells. A clonogenic assay showed that most granulocyte-macrophage colony-forming cells (GM-CFC) were present in CD34+KIThigh populations, whereas erythroid burst-forming cells (BFU-E) were detected mainly in the CD34+KITlow population. CD34+-KIT– fraction contained a small number of BFU-E. Morphologic analysis showed that blast-like cells were more enriched in the CD34+KITlow fraction. KITlow cells contained CD34+CD38– cells that were considered to be very primitive progenitor cells, as determined by a replating assay. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell functions by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At week 2, more CFC recovered from the culture in the fraction initiated with a CD34+KIThigh population. However, more LTC-IC were present during weeks 5 to 9 in the CD34+KITlow population. These results indicate that primitive progenitors are more enriched in the KITlow population and that the KIThigh population contains many GM-committed progenitor cells. We also showed that anti-KIT MoAb inhibited the ability of CD34+ cells to generate CFC on the stromal layer in the LTC system. This suppressive effect was more evident in the generation of BFU-E by CD34+KITlow cells. Moreover, we confirmed that CD34+KIThigh cells emerged from CD34+KITlow cells during coculture with allogeneic stromal cells or from liquid culture in the presence of stem cell factor (SCF), interleukin-6, and erythropoietin. These results emphasize the pivotal role of the KIT and SCF interaction in hematopoiesis and indicate that KITlow cells are more primitive than KIThigh cells.
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.V82.11.3283.3283