Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO

Norio Komatsu, Masae Kunitama, Minami Yamada, Tetsuya Hagiwara, Takashi Kato, Hiroshi Miyazaki, Mitsuoki Eguchi, Masayuki Yamamoto, Yasusada Miura

研究成果: Article

81 引用 (Scopus)

抄録

UT-7 is a human megakaryoblastic leukemia cell line with absolute dependence on interleukin-3, granulocyte-macrophage colony-stimulating factor, or erythropoietin (EPO) for growth and survival. We investigated the effect of thrombopoietin (TPO), the ligand for the receptor encoded by c-mpl proto-oncogene, on the proliferation and differentiation of UT-7 and its sublines. We found that UT-7/GM, which is a subline of UT-7, but neither UT- 7 nor UT-7/EPO, can proliferate in response to TPO. The subline, UT-7/TPO, was established from UT-7/GM by culture at lower concentrations of TPO. UT- 7/TPO cells had morphologically mature megakaryocytic characteristics such as developed demarcation membrane in the cytoplasm and multinucleated appearance. This was also confirmed by the high expression of platelet factor-4 and glycoprotein IIb at the mRNA levels and by the high level of DNA content. UT-7/TPO can be maintained by TPO alone, with a doubling time of 24 hours in log growth phase. In the absence of TPO, the majority of the cells died within a few days. Thus, UT-7/TPO has an absolute dependence on TPO for growth and survival and has mature megakaryocytic features. The mRNA for c- mpl was detected in UT-7/TPO and, to a lesser degree, in UT-7/GM. The mRNA level of NFE2 p45, reported to be an erythroid-specific transcription factor, was upregulated in UT-7/TPO, whereas it was down-regulated in the erythroid subline, UT-7/EPO. There were no significant differences in GATA1 and GATA-2 mRNA levels among UT-7 and its sublines. Not only EPO but also TPO induced the tyrosine phosphorylation of JAK2 tyrosine kinase and STAT5-related protein, These findings indicate that UT-7/TPO would be a useful model with which to analyze the gene regulation of megakaryocytic maturation-associated proteins and to study the specific actions of TPO.

元の言語English
ページ(範囲)4552-4560
ページ数9
ジャーナルBlood
87
発行部数11
出版物ステータスPublished - 1996 6 1
外部発表Yes

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Thrombopoietin
Cells
Cell Line
Erythropoietin
Messenger RNA
Growth
Thrombopoietin Receptors
STAT5 Transcription Factor
Platelet Factor 4
Survival
Proto-Oncogenes
Interleukin-3
Phosphorylation
Granulocyte-Macrophage Colony-Stimulating Factor
Protein-Tyrosine Kinases

ASJC Scopus subject areas

  • Hematology

これを引用

Komatsu, N., Kunitama, M., Yamada, M., Hagiwara, T., Kato, T., Miyazaki, H., ... Miura, Y. (1996). Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO. Blood, 87(11), 4552-4560.

Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO. / Komatsu, Norio; Kunitama, Masae; Yamada, Minami; Hagiwara, Tetsuya; Kato, Takashi; Miyazaki, Hiroshi; Eguchi, Mitsuoki; Yamamoto, Masayuki; Miura, Yasusada.

:: Blood, 巻 87, 番号 11, 01.06.1996, p. 4552-4560.

研究成果: Article

Komatsu, N, Kunitama, M, Yamada, M, Hagiwara, T, Kato, T, Miyazaki, H, Eguchi, M, Yamamoto, M & Miura, Y 1996, 'Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO', Blood, 巻. 87, 番号 11, pp. 4552-4560.
Komatsu N, Kunitama M, Yamada M, Hagiwara T, Kato T, Miyazaki H その他. Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO. Blood. 1996 6 1;87(11):4552-4560.
Komatsu, Norio ; Kunitama, Masae ; Yamada, Minami ; Hagiwara, Tetsuya ; Kato, Takashi ; Miyazaki, Hiroshi ; Eguchi, Mitsuoki ; Yamamoto, Masayuki ; Miura, Yasusada. / Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO. :: Blood. 1996 ; 巻 87, 番号 11. pp. 4552-4560.
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abstract = "UT-7 is a human megakaryoblastic leukemia cell line with absolute dependence on interleukin-3, granulocyte-macrophage colony-stimulating factor, or erythropoietin (EPO) for growth and survival. We investigated the effect of thrombopoietin (TPO), the ligand for the receptor encoded by c-mpl proto-oncogene, on the proliferation and differentiation of UT-7 and its sublines. We found that UT-7/GM, which is a subline of UT-7, but neither UT- 7 nor UT-7/EPO, can proliferate in response to TPO. The subline, UT-7/TPO, was established from UT-7/GM by culture at lower concentrations of TPO. UT- 7/TPO cells had morphologically mature megakaryocytic characteristics such as developed demarcation membrane in the cytoplasm and multinucleated appearance. This was also confirmed by the high expression of platelet factor-4 and glycoprotein IIb at the mRNA levels and by the high level of DNA content. UT-7/TPO can be maintained by TPO alone, with a doubling time of 24 hours in log growth phase. In the absence of TPO, the majority of the cells died within a few days. Thus, UT-7/TPO has an absolute dependence on TPO for growth and survival and has mature megakaryocytic features. The mRNA for c- mpl was detected in UT-7/TPO and, to a lesser degree, in UT-7/GM. The mRNA level of NFE2 p45, reported to be an erythroid-specific transcription factor, was upregulated in UT-7/TPO, whereas it was down-regulated in the erythroid subline, UT-7/EPO. There were no significant differences in GATA1 and GATA-2 mRNA levels among UT-7 and its sublines. Not only EPO but also TPO induced the tyrosine phosphorylation of JAK2 tyrosine kinase and STAT5-related protein, These findings indicate that UT-7/TPO would be a useful model with which to analyze the gene regulation of megakaryocytic maturation-associated proteins and to study the specific actions of TPO.",
author = "Norio Komatsu and Masae Kunitama and Minami Yamada and Tetsuya Hagiwara and Takashi Kato and Hiroshi Miyazaki and Mitsuoki Eguchi and Masayuki Yamamoto and Yasusada Miura",
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T1 - Establishment and characterization of the thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO

AU - Komatsu, Norio

AU - Kunitama, Masae

AU - Yamada, Minami

AU - Hagiwara, Tetsuya

AU - Kato, Takashi

AU - Miyazaki, Hiroshi

AU - Eguchi, Mitsuoki

AU - Yamamoto, Masayuki

AU - Miura, Yasusada

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N2 - UT-7 is a human megakaryoblastic leukemia cell line with absolute dependence on interleukin-3, granulocyte-macrophage colony-stimulating factor, or erythropoietin (EPO) for growth and survival. We investigated the effect of thrombopoietin (TPO), the ligand for the receptor encoded by c-mpl proto-oncogene, on the proliferation and differentiation of UT-7 and its sublines. We found that UT-7/GM, which is a subline of UT-7, but neither UT- 7 nor UT-7/EPO, can proliferate in response to TPO. The subline, UT-7/TPO, was established from UT-7/GM by culture at lower concentrations of TPO. UT- 7/TPO cells had morphologically mature megakaryocytic characteristics such as developed demarcation membrane in the cytoplasm and multinucleated appearance. This was also confirmed by the high expression of platelet factor-4 and glycoprotein IIb at the mRNA levels and by the high level of DNA content. UT-7/TPO can be maintained by TPO alone, with a doubling time of 24 hours in log growth phase. In the absence of TPO, the majority of the cells died within a few days. Thus, UT-7/TPO has an absolute dependence on TPO for growth and survival and has mature megakaryocytic features. The mRNA for c- mpl was detected in UT-7/TPO and, to a lesser degree, in UT-7/GM. The mRNA level of NFE2 p45, reported to be an erythroid-specific transcription factor, was upregulated in UT-7/TPO, whereas it was down-regulated in the erythroid subline, UT-7/EPO. There were no significant differences in GATA1 and GATA-2 mRNA levels among UT-7 and its sublines. Not only EPO but also TPO induced the tyrosine phosphorylation of JAK2 tyrosine kinase and STAT5-related protein, These findings indicate that UT-7/TPO would be a useful model with which to analyze the gene regulation of megakaryocytic maturation-associated proteins and to study the specific actions of TPO.

AB - UT-7 is a human megakaryoblastic leukemia cell line with absolute dependence on interleukin-3, granulocyte-macrophage colony-stimulating factor, or erythropoietin (EPO) for growth and survival. We investigated the effect of thrombopoietin (TPO), the ligand for the receptor encoded by c-mpl proto-oncogene, on the proliferation and differentiation of UT-7 and its sublines. We found that UT-7/GM, which is a subline of UT-7, but neither UT- 7 nor UT-7/EPO, can proliferate in response to TPO. The subline, UT-7/TPO, was established from UT-7/GM by culture at lower concentrations of TPO. UT- 7/TPO cells had morphologically mature megakaryocytic characteristics such as developed demarcation membrane in the cytoplasm and multinucleated appearance. This was also confirmed by the high expression of platelet factor-4 and glycoprotein IIb at the mRNA levels and by the high level of DNA content. UT-7/TPO can be maintained by TPO alone, with a doubling time of 24 hours in log growth phase. In the absence of TPO, the majority of the cells died within a few days. Thus, UT-7/TPO has an absolute dependence on TPO for growth and survival and has mature megakaryocytic features. The mRNA for c- mpl was detected in UT-7/TPO and, to a lesser degree, in UT-7/GM. The mRNA level of NFE2 p45, reported to be an erythroid-specific transcription factor, was upregulated in UT-7/TPO, whereas it was down-regulated in the erythroid subline, UT-7/EPO. There were no significant differences in GATA1 and GATA-2 mRNA levels among UT-7 and its sublines. Not only EPO but also TPO induced the tyrosine phosphorylation of JAK2 tyrosine kinase and STAT5-related protein, These findings indicate that UT-7/TPO would be a useful model with which to analyze the gene regulation of megakaryocytic maturation-associated proteins and to study the specific actions of TPO.

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