Growth and differentiation of a human megakaryoblastic cell line, CMK

N. Komatsu, T. Suda, M. Moroi, N. Tokuyama, Y. Sakata, M. Okada, T. Nishida, Y. Hirai, T. Sato, A. Fuse, Y. Miura

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Abstract

Recently, a human megakaryoblastic cell line, CMK, was established from the peripheral blood of a megakaryoblastic leukemia patient with Down syndrome. Using this cell line, we studied the proliferation and differentiation of megakaryotic cells in the presence of highly purified human hematopoietic factors and phorbol 12-myristate-13-acetate (PMA). In a methylcellulose culture system, interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitated colony formation by CMK cells in a dose-dependent manner. The maximum stimulating doses of these factors were 10 and 200 U/mL, respectively. These concentrations were comparable to those that stimulate activity in normal hematopoietic cells. In contrast, granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF), and erythropoietin (EPO) had no effects on the colony formation of CMK cells. In a liquid culture system, 20% of the CMK cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hematopoietic factors, whereas 40% of the cells expressed GPIIb/IIIa with the addition of IL-3 and GM-CSF. EPO also slightly enhanced expression of GPIIb/IIIa. On the other hand, PMA inhibited growth of CMK cells and induced most of them to express the GPIIb/IIIa antigen. Furthermore, PMA induced CMK cells to produce growth activity toward new inocula of CMK cells. This growth factor (GF) contained colony-stimulating activity (CSA) in normal bone marrow (BM) cells. The activity was believed to be attributable mainly to GM-CSF, since 64% of this activity was neutralized by anti-GM-CSF antibodies and a transcript of GM-CSF was detected in mRNA from PMA-treated CMK cells by Northern blot analysis. These observations suggest that GM-CSF, as well as IL-3, should play an important role in megakaryocytopoiesis.

Original languageEnglish
Pages (from-to)42-48
Number of pages7
JournalBlood
Volume74
Issue number1
Publication statusPublished - 1989

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Granulocyte-Macrophage Colony-Stimulating Factor
Cells
Cell Line
Interleukin-3
Acetates
Growth
Erythropoietin
Antigens
Platelet Glycoprotein GPIIb-IIIa Complex
Methylcellulose
Macrophage Colony-Stimulating Factor
Human engineering
Thrombopoiesis
Intercellular Signaling Peptides and Proteins
Bone
Blood
Granulocyte Colony-Stimulating Factor
Down Syndrome
Bone Marrow Cells
Northern Blotting

ASJC Scopus subject areas

  • Hematology

Cite this

Komatsu, N., Suda, T., Moroi, M., Tokuyama, N., Sakata, Y., Okada, M., ... Miura, Y. (1989). Growth and differentiation of a human megakaryoblastic cell line, CMK. Blood, 74(1), 42-48.

Growth and differentiation of a human megakaryoblastic cell line, CMK. / Komatsu, N.; Suda, T.; Moroi, M.; Tokuyama, N.; Sakata, Y.; Okada, M.; Nishida, T.; Hirai, Y.; Sato, T.; Fuse, A.; Miura, Y.

In: Blood, Vol. 74, No. 1, 1989, p. 42-48.

Research output: Contribution to journalArticle

Komatsu, N, Suda, T, Moroi, M, Tokuyama, N, Sakata, Y, Okada, M, Nishida, T, Hirai, Y, Sato, T, Fuse, A & Miura, Y 1989, 'Growth and differentiation of a human megakaryoblastic cell line, CMK', Blood, vol. 74, no. 1, pp. 42-48.
Komatsu N, Suda T, Moroi M, Tokuyama N, Sakata Y, Okada M et al. Growth and differentiation of a human megakaryoblastic cell line, CMK. Blood. 1989;74(1):42-48.
Komatsu, N. ; Suda, T. ; Moroi, M. ; Tokuyama, N. ; Sakata, Y. ; Okada, M. ; Nishida, T. ; Hirai, Y. ; Sato, T. ; Fuse, A. ; Miura, Y. / Growth and differentiation of a human megakaryoblastic cell line, CMK. In: Blood. 1989 ; Vol. 74, No. 1. pp. 42-48.
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abstract = "Recently, a human megakaryoblastic cell line, CMK, was established from the peripheral blood of a megakaryoblastic leukemia patient with Down syndrome. Using this cell line, we studied the proliferation and differentiation of megakaryotic cells in the presence of highly purified human hematopoietic factors and phorbol 12-myristate-13-acetate (PMA). In a methylcellulose culture system, interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitated colony formation by CMK cells in a dose-dependent manner. The maximum stimulating doses of these factors were 10 and 200 U/mL, respectively. These concentrations were comparable to those that stimulate activity in normal hematopoietic cells. In contrast, granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF), and erythropoietin (EPO) had no effects on the colony formation of CMK cells. In a liquid culture system, 20{\%} of the CMK cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hematopoietic factors, whereas 40{\%} of the cells expressed GPIIb/IIIa with the addition of IL-3 and GM-CSF. EPO also slightly enhanced expression of GPIIb/IIIa. On the other hand, PMA inhibited growth of CMK cells and induced most of them to express the GPIIb/IIIa antigen. Furthermore, PMA induced CMK cells to produce growth activity toward new inocula of CMK cells. This growth factor (GF) contained colony-stimulating activity (CSA) in normal bone marrow (BM) cells. The activity was believed to be attributable mainly to GM-CSF, since 64{\%} of this activity was neutralized by anti-GM-CSF antibodies and a transcript of GM-CSF was detected in mRNA from PMA-treated CMK cells by Northern blot analysis. These observations suggest that GM-CSF, as well as IL-3, should play an important role in megakaryocytopoiesis.",
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T1 - Growth and differentiation of a human megakaryoblastic cell line, CMK

AU - Komatsu, N.

AU - Suda, T.

AU - Moroi, M.

AU - Tokuyama, N.

AU - Sakata, Y.

AU - Okada, M.

AU - Nishida, T.

AU - Hirai, Y.

AU - Sato, T.

AU - Fuse, A.

AU - Miura, Y.

PY - 1989

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N2 - Recently, a human megakaryoblastic cell line, CMK, was established from the peripheral blood of a megakaryoblastic leukemia patient with Down syndrome. Using this cell line, we studied the proliferation and differentiation of megakaryotic cells in the presence of highly purified human hematopoietic factors and phorbol 12-myristate-13-acetate (PMA). In a methylcellulose culture system, interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitated colony formation by CMK cells in a dose-dependent manner. The maximum stimulating doses of these factors were 10 and 200 U/mL, respectively. These concentrations were comparable to those that stimulate activity in normal hematopoietic cells. In contrast, granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF), and erythropoietin (EPO) had no effects on the colony formation of CMK cells. In a liquid culture system, 20% of the CMK cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hematopoietic factors, whereas 40% of the cells expressed GPIIb/IIIa with the addition of IL-3 and GM-CSF. EPO also slightly enhanced expression of GPIIb/IIIa. On the other hand, PMA inhibited growth of CMK cells and induced most of them to express the GPIIb/IIIa antigen. Furthermore, PMA induced CMK cells to produce growth activity toward new inocula of CMK cells. This growth factor (GF) contained colony-stimulating activity (CSA) in normal bone marrow (BM) cells. The activity was believed to be attributable mainly to GM-CSF, since 64% of this activity was neutralized by anti-GM-CSF antibodies and a transcript of GM-CSF was detected in mRNA from PMA-treated CMK cells by Northern blot analysis. These observations suggest that GM-CSF, as well as IL-3, should play an important role in megakaryocytopoiesis.

AB - Recently, a human megakaryoblastic cell line, CMK, was established from the peripheral blood of a megakaryoblastic leukemia patient with Down syndrome. Using this cell line, we studied the proliferation and differentiation of megakaryotic cells in the presence of highly purified human hematopoietic factors and phorbol 12-myristate-13-acetate (PMA). In a methylcellulose culture system, interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitated colony formation by CMK cells in a dose-dependent manner. The maximum stimulating doses of these factors were 10 and 200 U/mL, respectively. These concentrations were comparable to those that stimulate activity in normal hematopoietic cells. In contrast, granulocyte-colony stimulating factor (G-CSF), macrophage-colony stimulating factor (M-CSF), and erythropoietin (EPO) had no effects on the colony formation of CMK cells. In a liquid culture system, 20% of the CMK cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hematopoietic factors, whereas 40% of the cells expressed GPIIb/IIIa with the addition of IL-3 and GM-CSF. EPO also slightly enhanced expression of GPIIb/IIIa. On the other hand, PMA inhibited growth of CMK cells and induced most of them to express the GPIIb/IIIa antigen. Furthermore, PMA induced CMK cells to produce growth activity toward new inocula of CMK cells. This growth factor (GF) contained colony-stimulating activity (CSA) in normal bone marrow (BM) cells. The activity was believed to be attributable mainly to GM-CSF, since 64% of this activity was neutralized by anti-GM-CSF antibodies and a transcript of GM-CSF was detected in mRNA from PMA-treated CMK cells by Northern blot analysis. These observations suggest that GM-CSF, as well as IL-3, should play an important role in megakaryocytopoiesis.

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