Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells

Yuji Gunji, Mitsuru Nakamura, Hisao Osawa, Kazunari Nagayoshi, Hiromitsu Nakauchi, Yasusada Miura, Masayoshi Yanagisawa, Toshio Suda

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Pages (from-to)3283-3289
Number of pages7
JournalBlood
Volume82
Issue number11
Publication statusPublished - 1993 Dec 1

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Chlorofluorocarbons
Stem Cell Factor
Hematopoietic Stem Cells
Cell culture
Assays
Monoclonal Antibodies
Macrophages
Oncogene Proteins
Erythropoietin
Interleukin-6
Liquids
Erythroid Precursor Cells
Population
Stem Cells
Stromal Cells
Cell Culture Techniques
Proto-Oncogenes
Hematopoiesis
Coculture Techniques
Granulocytes

ASJC Scopus subject areas

  • Hematology

Cite this

Gunji, Y., Nakamura, M., Osawa, H., Nagayoshi, K., Nakauchi, H., Miura, Y., ... Suda, T. (1993). Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells. Blood, 82(11), 3283-3289.

Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells. / Gunji, Yuji; Nakamura, Mitsuru; Osawa, Hisao; Nagayoshi, Kazunari; Nakauchi, Hiromitsu; Miura, Yasusada; Yanagisawa, Masayoshi; Suda, Toshio.

In: Blood, Vol. 82, No. 11, 01.12.1993, p. 3283-3289.

Research output: Contribution to journalArticle

Gunji, Y, Nakamura, M, Osawa, H, Nagayoshi, K, Nakauchi, H, Miura, Y, Yanagisawa, M & Suda, T 1993, 'Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells', Blood, vol. 82, no. 11, pp. 3283-3289.
Gunji Y, Nakamura M, Osawa H, Nagayoshi K, Nakauchi H, Miura Y et al. Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells. Blood. 1993 Dec 1;82(11):3283-3289.
Gunji, Yuji ; Nakamura, Mitsuru ; Osawa, Hisao ; Nagayoshi, Kazunari ; Nakauchi, Hiromitsu ; Miura, Yasusada ; Yanagisawa, Masayoshi ; Suda, Toshio. / Human primitive hematopoietic progenitor cells are more enriched in KITlow cells than in KIThigh cells. In: Blood. 1993 ; Vol. 82, No. 11. pp. 3283-3289.
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abstract = "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.",
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AU - Gunji, Yuji

AU - Nakamura, Mitsuru

AU - Osawa, Hisao

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AU - Nakauchi, Hiromitsu

AU - Miura, Yasusada

AU - Yanagisawa, Masayoshi

AU - Suda, Toshio

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N2 - 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.

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