Synergistic effect of IL-3 and IL-6 on highly enriched murine hemopoietic progenitors

S. Okada, H. Nakauchi, K. Nagayoshi, M. Nakamura, Y. Miura, T. Suda

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Abstract

It has been reported that interleukin 6 (IL-6) acts on hemopoietic stem cells in synergism with interleukin 3 (IL-3), but it has not yet been clarified whether IL-6 acts directly on the stem cells or not. To investigate the mechanism of the synergism between IL-3 and IL-6, we sorted hemopoietic stem cells from untreated murine bone marrow cells using a two-laser fluorescence-activated cell sorter (FACS). Cells negative for the lymphohemopoietic lineage (lineage-negative, Lin-), with a high affinity to wheat germ agglutinin (WGA+), and showing a low expression of Thy-1 antigen (Thy-1(low)) were sorted and analyzed by in vitro and in vivo colony formation. This fraction was 0.4% of the total mononuclear bone marrow cells. Approximately 25% of these Lin-WGA+Thy-1(low) cells showed in vitro colony formation, whereas approximately 1% of them formed day-8 and day-12 spleen colonies. Thus, it appears that the Lin-WGA+Thy1(low) cells were a highly enriched stem cell population. By FACS clone sorting, single cells were isolated from the enriched stem cell fraction and cultured in semisolid or liquid culture systems. Addition of IL-6 to methylcellulose medium containing IL-3 did not significantly increase the number of colonies. It is thus suggested that the target cells of IL-3 and IL-6 are the same as those of IL-3. The secondary colony-forming ability of primary colonies that developed in the presence of IL-6 and IL-3 was higher than that of colonies formed in the presence of IL-3 alone. In correspondence with this finding, the numbers of myeloid colonies and spleen colony-forming units (CFU-S) were increased by the incubation of these sorted cells for 7 days with IL-6 and IL-3 when compared with the effect of IL-3 alone. Therefore, it is concluded that IL-6 acts directly on hemopoietic stem cells to enhance their proliferation.

Original languageEnglish
Pages (from-to)546-551
Number of pages6
JournalExperimental Hematology
Volume20
Issue number5
Publication statusPublished - 1992

Fingerprint

Interleukin-3
Interleukin-6
Stem Cells
Bone Marrow Cells
Spleen
Fluorescence
Thy-1 Antigens
Wheat Germ Agglutinins
Methylcellulose
Lasers
Clone Cells

Keywords

  • enriched hemopoietic cells
  • interleukin 3
  • interleukin 6

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Okada, S., Nakauchi, H., Nagayoshi, K., Nakamura, M., Miura, Y., & Suda, T. (1992). Synergistic effect of IL-3 and IL-6 on highly enriched murine hemopoietic progenitors. Experimental Hematology, 20(5), 546-551.

Synergistic effect of IL-3 and IL-6 on highly enriched murine hemopoietic progenitors. / Okada, S.; Nakauchi, H.; Nagayoshi, K.; Nakamura, M.; Miura, Y.; Suda, T.

In: Experimental Hematology, Vol. 20, No. 5, 1992, p. 546-551.

Research output: Contribution to journalArticle

Okada, S, Nakauchi, H, Nagayoshi, K, Nakamura, M, Miura, Y & Suda, T 1992, 'Synergistic effect of IL-3 and IL-6 on highly enriched murine hemopoietic progenitors', Experimental Hematology, vol. 20, no. 5, pp. 546-551.
Okada S, Nakauchi H, Nagayoshi K, Nakamura M, Miura Y, Suda T. Synergistic effect of IL-3 and IL-6 on highly enriched murine hemopoietic progenitors. Experimental Hematology. 1992;20(5):546-551.
Okada, S. ; Nakauchi, H. ; Nagayoshi, K. ; Nakamura, M. ; Miura, Y. ; Suda, T. / Synergistic effect of IL-3 and IL-6 on highly enriched murine hemopoietic progenitors. In: Experimental Hematology. 1992 ; Vol. 20, No. 5. pp. 546-551.
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abstract = "It has been reported that interleukin 6 (IL-6) acts on hemopoietic stem cells in synergism with interleukin 3 (IL-3), but it has not yet been clarified whether IL-6 acts directly on the stem cells or not. To investigate the mechanism of the synergism between IL-3 and IL-6, we sorted hemopoietic stem cells from untreated murine bone marrow cells using a two-laser fluorescence-activated cell sorter (FACS). Cells negative for the lymphohemopoietic lineage (lineage-negative, Lin-), with a high affinity to wheat germ agglutinin (WGA+), and showing a low expression of Thy-1 antigen (Thy-1(low)) were sorted and analyzed by in vitro and in vivo colony formation. This fraction was 0.4{\%} of the total mononuclear bone marrow cells. Approximately 25{\%} of these Lin-WGA+Thy-1(low) cells showed in vitro colony formation, whereas approximately 1{\%} of them formed day-8 and day-12 spleen colonies. Thus, it appears that the Lin-WGA+Thy1(low) cells were a highly enriched stem cell population. By FACS clone sorting, single cells were isolated from the enriched stem cell fraction and cultured in semisolid or liquid culture systems. Addition of IL-6 to methylcellulose medium containing IL-3 did not significantly increase the number of colonies. It is thus suggested that the target cells of IL-3 and IL-6 are the same as those of IL-3. The secondary colony-forming ability of primary colonies that developed in the presence of IL-6 and IL-3 was higher than that of colonies formed in the presence of IL-3 alone. In correspondence with this finding, the numbers of myeloid colonies and spleen colony-forming units (CFU-S) were increased by the incubation of these sorted cells for 7 days with IL-6 and IL-3 when compared with the effect of IL-3 alone. Therefore, it is concluded that IL-6 acts directly on hemopoietic stem cells to enhance their proliferation.",
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AB - It has been reported that interleukin 6 (IL-6) acts on hemopoietic stem cells in synergism with interleukin 3 (IL-3), but it has not yet been clarified whether IL-6 acts directly on the stem cells or not. To investigate the mechanism of the synergism between IL-3 and IL-6, we sorted hemopoietic stem cells from untreated murine bone marrow cells using a two-laser fluorescence-activated cell sorter (FACS). Cells negative for the lymphohemopoietic lineage (lineage-negative, Lin-), with a high affinity to wheat germ agglutinin (WGA+), and showing a low expression of Thy-1 antigen (Thy-1(low)) were sorted and analyzed by in vitro and in vivo colony formation. This fraction was 0.4% of the total mononuclear bone marrow cells. Approximately 25% of these Lin-WGA+Thy-1(low) cells showed in vitro colony formation, whereas approximately 1% of them formed day-8 and day-12 spleen colonies. Thus, it appears that the Lin-WGA+Thy1(low) cells were a highly enriched stem cell population. By FACS clone sorting, single cells were isolated from the enriched stem cell fraction and cultured in semisolid or liquid culture systems. Addition of IL-6 to methylcellulose medium containing IL-3 did not significantly increase the number of colonies. It is thus suggested that the target cells of IL-3 and IL-6 are the same as those of IL-3. The secondary colony-forming ability of primary colonies that developed in the presence of IL-6 and IL-3 was higher than that of colonies formed in the presence of IL-3 alone. In correspondence with this finding, the numbers of myeloid colonies and spleen colony-forming units (CFU-S) were increased by the incubation of these sorted cells for 7 days with IL-6 and IL-3 when compared with the effect of IL-3 alone. Therefore, it is concluded that IL-6 acts directly on hemopoietic stem cells to enhance their proliferation.

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