Characterization of monocyte-macrophage-lineage cells induced from CD34+ bone marrow cells in vitro

Kyoko Suzuki; Nobutaka Kiyokawa; Tomoko Taguchi; Hisami Takenouchi; Masahiro Saito; Toshiaki Shimizu; Hajime Okita; Junichiro Fujimoto

doi:10.1532/IJH97.06213

Characterization of monocyte-macrophage-lineage cells induced from CD34⁺ bone marrow cells in vitro

Kyoko Suzuki, Nobutaka Kiyokawa, Tomoko Taguchi, Hisami Takenouchi, Masahiro Saito, Toshiaki Shimizu, Hajime Okita, Junichiro Fujimoto

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

We characterized the expression of cell surface antigens and cytokine-secreting ability of monocyte-macrophage-lineage cells induced in vitro from CD34⁺ bone marrow cells. After cultivation for 3 weeks, we observed 2 distinct cell fractions: a floating small, round cell fraction and an adherent large, protruding cell fraction. Both cell fractions expressed myelocyte-monocyte-lineage antigens, but mature-macrophage markers such as CD206 were expressed only by the adherent cells. An assessment of cells cultured for 5 weeks revealed spontaneous secretion of interleukin 8 (IL-8) and IL-6, and lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNF-α) secretion in both fractions, but only the adherent cell fraction secreted IL-10 after LPS stimulation. In contrast, both fractions of cells cultured for 3 weeks spontaneously secreted low levels of IL-8, but none of the other cytokines. Upon LPS stimulation, the cells secreted IL-6 and TNF-α, but not IL-10. We also assessed the effect of granulocyte colony-stimulating factor (G-CSF) pretreatment on TNF-α secretion by each cell fraction and found that G-CSF reduced TNF-α secretion only in the adherent fraction of cells cultured for 3 weeks. Monocyte-macrophage-lineage cells induced in vitro should provide an ideal model for functional analysis of monocyte-macrophage cells.

Original language	English
Pages (from-to)	384-389
Number of pages	6
Journal	International journal of hematology
Volume	85
Issue number	5
DOIs	https://doi.org/10.1532/IJH97.06213
Publication status	Published - 2007 Jun
Externally published	Yes

Keywords

Antigen
Cytokine
Expression
Monocyte-macrophage lineage

ASJC Scopus subject areas

Hematology

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Characterization of monocyte-macrophage-lineage cells induced from CD34⁺ bone marrow cells in vitro. / Suzuki, Kyoko; Kiyokawa, Nobutaka; Taguchi, Tomoko; Takenouchi, Hisami; Saito, Masahiro; Shimizu, Toshiaki; Okita, Hajime; Fujimoto, Junichiro.

In: International journal of hematology, Vol. 85, No. 5, 06.2007, p. 384-389.

Research output: Contribution to journal › Article › peer-review

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title = "Characterization of monocyte-macrophage-lineage cells induced from CD34+ bone marrow cells in vitro",

abstract = "We characterized the expression of cell surface antigens and cytokine-secreting ability of monocyte-macrophage-lineage cells induced in vitro from CD34+ bone marrow cells. After cultivation for 3 weeks, we observed 2 distinct cell fractions: a floating small, round cell fraction and an adherent large, protruding cell fraction. Both cell fractions expressed myelocyte-monocyte-lineage antigens, but mature-macrophage markers such as CD206 were expressed only by the adherent cells. An assessment of cells cultured for 5 weeks revealed spontaneous secretion of interleukin 8 (IL-8) and IL-6, and lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNF-α) secretion in both fractions, but only the adherent cell fraction secreted IL-10 after LPS stimulation. In contrast, both fractions of cells cultured for 3 weeks spontaneously secreted low levels of IL-8, but none of the other cytokines. Upon LPS stimulation, the cells secreted IL-6 and TNF-α, but not IL-10. We also assessed the effect of granulocyte colony-stimulating factor (G-CSF) pretreatment on TNF-α secretion by each cell fraction and found that G-CSF reduced TNF-α secretion only in the adherent fraction of cells cultured for 3 weeks. Monocyte-macrophage-lineage cells induced in vitro should provide an ideal model for functional analysis of monocyte-macrophage cells.",

keywords = "Antigen, Cytokine, Expression, Monocyte-macrophage lineage",

author = "Kyoko Suzuki and Nobutaka Kiyokawa and Tomoko Taguchi and Hisami Takenouchi and Masahiro Saito and Toshiaki Shimizu and Hajime Okita and Junichiro Fujimoto",

note = "Funding Information: This work was supported in part by Health and Labour Sciences Research Grants; a grant for Child Health and Development from the Ministry of Health, Labour and Welfare of Japan; and a grant from the Japan Health Sciences Foundation for Research on Health Sciences Focusing on Drug Innovation, JSPS. This work was supported by KAKENHI 17591131 and 18790751, the Budget for Nuclear Research of the Ministry of Education, Culture, Sports, Science and Technology, based on screening and counseling by the Atomic Energy Commission. This work was also supported by CREST, JST, and a grant from the Charitable Trust Japan Leukemia Research Fund. We thank S. Yamauchi for excellent secretarial work.",

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AU - Suzuki, Kyoko

AU - Kiyokawa, Nobutaka

AU - Taguchi, Tomoko

AU - Takenouchi, Hisami

AU - Saito, Masahiro

AU - Shimizu, Toshiaki

AU - Okita, Hajime

AU - Fujimoto, Junichiro

N1 - Funding Information: This work was supported in part by Health and Labour Sciences Research Grants; a grant for Child Health and Development from the Ministry of Health, Labour and Welfare of Japan; and a grant from the Japan Health Sciences Foundation for Research on Health Sciences Focusing on Drug Innovation, JSPS. This work was supported by KAKENHI 17591131 and 18790751, the Budget for Nuclear Research of the Ministry of Education, Culture, Sports, Science and Technology, based on screening and counseling by the Atomic Energy Commission. This work was also supported by CREST, JST, and a grant from the Charitable Trust Japan Leukemia Research Fund. We thank S. Yamauchi for excellent secretarial work.

PY - 2007/6

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N2 - We characterized the expression of cell surface antigens and cytokine-secreting ability of monocyte-macrophage-lineage cells induced in vitro from CD34+ bone marrow cells. After cultivation for 3 weeks, we observed 2 distinct cell fractions: a floating small, round cell fraction and an adherent large, protruding cell fraction. Both cell fractions expressed myelocyte-monocyte-lineage antigens, but mature-macrophage markers such as CD206 were expressed only by the adherent cells. An assessment of cells cultured for 5 weeks revealed spontaneous secretion of interleukin 8 (IL-8) and IL-6, and lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNF-α) secretion in both fractions, but only the adherent cell fraction secreted IL-10 after LPS stimulation. In contrast, both fractions of cells cultured for 3 weeks spontaneously secreted low levels of IL-8, but none of the other cytokines. Upon LPS stimulation, the cells secreted IL-6 and TNF-α, but not IL-10. We also assessed the effect of granulocyte colony-stimulating factor (G-CSF) pretreatment on TNF-α secretion by each cell fraction and found that G-CSF reduced TNF-α secretion only in the adherent fraction of cells cultured for 3 weeks. Monocyte-macrophage-lineage cells induced in vitro should provide an ideal model for functional analysis of monocyte-macrophage cells.

AB - We characterized the expression of cell surface antigens and cytokine-secreting ability of monocyte-macrophage-lineage cells induced in vitro from CD34+ bone marrow cells. After cultivation for 3 weeks, we observed 2 distinct cell fractions: a floating small, round cell fraction and an adherent large, protruding cell fraction. Both cell fractions expressed myelocyte-monocyte-lineage antigens, but mature-macrophage markers such as CD206 were expressed only by the adherent cells. An assessment of cells cultured for 5 weeks revealed spontaneous secretion of interleukin 8 (IL-8) and IL-6, and lipopolysaccharide (LPS)-induced tumor necrosis factor α (TNF-α) secretion in both fractions, but only the adherent cell fraction secreted IL-10 after LPS stimulation. In contrast, both fractions of cells cultured for 3 weeks spontaneously secreted low levels of IL-8, but none of the other cytokines. Upon LPS stimulation, the cells secreted IL-6 and TNF-α, but not IL-10. We also assessed the effect of granulocyte colony-stimulating factor (G-CSF) pretreatment on TNF-α secretion by each cell fraction and found that G-CSF reduced TNF-α secretion only in the adherent fraction of cells cultured for 3 weeks. Monocyte-macrophage-lineage cells induced in vitro should provide an ideal model for functional analysis of monocyte-macrophage cells.

KW - Antigen

KW - Cytokine

KW - Expression

KW - Monocyte-macrophage lineage

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