Human osteoclast-like cells are formed from peripheral blood mononuclear cells in a coculture with SaOS-2 cells transfected with the parathyroid hormone (PTH)/PTH-related protein receptor gene

Kenichiro Matsuzaki, Kazuhiko Katayama, Yasuyuki Takahashi, Ichiro Nakamura, Nobuyuki Udagawa, Taro Tsurukai, Ryuichi Nishinakamura, Yoshiaki Toyama, Yutaka Yabe, Masayuki Hori, Naoyuki Takahashi, Tatsuo Suda

Research output: Contribution to journalArticle

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Abstract

Subclones of the human osteosarcoma cell line SaOS-2 were established by transfecting with an expression vector containing the human PTH/PTH-related protein (PTHrP) receptor, and their abilities to support osteoclast-like multinucleated cell (OCL) formation were examined in coculture with mouse or human hemopoietic cells. Of four subclones examined, SAOS-2/4 and SAOS-4/3 bound high levels of [125I]-PTH and produced a significant amount of cAMP in response to PTH. OCLs were formed in response to PTH in the cocultures of mouse bone marrow cells with either SaOS-2/4 cells or SAOS-4/3 cells. Human OCLs were also formed in response to PTH in the coculture of SaOS-4/3 cells and human peripheral blood mononuclear cells. Adding dexamethasone together with PTH greatly enhanced PTH-induced human OCL formation. Like mouse OCLs, human OCLs formed in response to PTH were tartrate-resistant acid phosphatase positive, expressed abundant calcitonin receptors and vitronectin receptors, and formed resorption pits on dentine slices. Other osteotropic factors such as 1α,25-dihydroxyvitamin D3, prostaglandin E2, and interleukin 6 plus soluble interleukin 6 receptors failed to induce mouse and human OCLs in cocultures with SAOS-4/3 cells. Both mouse and human OCL formation supported by SAOS-4/3 cells were inhibited by either adding an antibody against macrophage-colony stimulating factor or adding granulocyte/macrophage-colony stimulating factor. Thus, it is likely that human and mouse OCL formation supported by SaOS-4/3 cells are similarly regulated. These results indicate that the target cells of PTH for inducing osteoclast formation are osteoblast/stromal cells but not osteoclast progenitor cells in the coculture. This coculture model will be useful for investigating the abnormalities of osteoclast differentiation and function in human metabolic bone diseases.

Original languageEnglish
Pages (from-to)925-932
Number of pages8
JournalEndocrinology
Volume140
Issue number2
DOIs
Publication statusPublished - 1999

Fingerprint

Parathyroid Hormone-Related Protein
Osteoclasts
Coculture Techniques
Parathyroid Hormone
Blood Cells
Genes
Vitronectin Receptors
Calcitonin Receptors
Interleukin-6 Receptors
Macrophage Colony-Stimulating Factor
Calcitriol
Metabolic Bone Diseases
Osteosarcoma
Dentin
Granulocyte-Macrophage Colony-Stimulating Factor
Stromal Cells
Osteoblasts
Dinoprostone
Bone Marrow Cells
Dexamethasone

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Human osteoclast-like cells are formed from peripheral blood mononuclear cells in a coculture with SaOS-2 cells transfected with the parathyroid hormone (PTH)/PTH-related protein receptor gene. / Matsuzaki, Kenichiro; Katayama, Kazuhiko; Takahashi, Yasuyuki; Nakamura, Ichiro; Udagawa, Nobuyuki; Tsurukai, Taro; Nishinakamura, Ryuichi; Toyama, Yoshiaki; Yabe, Yutaka; Hori, Masayuki; Takahashi, Naoyuki; Suda, Tatsuo.

In: Endocrinology, Vol. 140, No. 2, 1999, p. 925-932.

Research output: Contribution to journalArticle

Matsuzaki, K, Katayama, K, Takahashi, Y, Nakamura, I, Udagawa, N, Tsurukai, T, Nishinakamura, R, Toyama, Y, Yabe, Y, Hori, M, Takahashi, N & Suda, T 1999, 'Human osteoclast-like cells are formed from peripheral blood mononuclear cells in a coculture with SaOS-2 cells transfected with the parathyroid hormone (PTH)/PTH-related protein receptor gene', Endocrinology, vol. 140, no. 2, pp. 925-932. https://doi.org/10.1210/en.140.2.925
Matsuzaki, Kenichiro ; Katayama, Kazuhiko ; Takahashi, Yasuyuki ; Nakamura, Ichiro ; Udagawa, Nobuyuki ; Tsurukai, Taro ; Nishinakamura, Ryuichi ; Toyama, Yoshiaki ; Yabe, Yutaka ; Hori, Masayuki ; Takahashi, Naoyuki ; Suda, Tatsuo. / Human osteoclast-like cells are formed from peripheral blood mononuclear cells in a coculture with SaOS-2 cells transfected with the parathyroid hormone (PTH)/PTH-related protein receptor gene. In: Endocrinology. 1999 ; Vol. 140, No. 2. pp. 925-932.
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abstract = "Subclones of the human osteosarcoma cell line SaOS-2 were established by transfecting with an expression vector containing the human PTH/PTH-related protein (PTHrP) receptor, and their abilities to support osteoclast-like multinucleated cell (OCL) formation were examined in coculture with mouse or human hemopoietic cells. Of four subclones examined, SAOS-2/4 and SAOS-4/3 bound high levels of [125I]-PTH and produced a significant amount of cAMP in response to PTH. OCLs were formed in response to PTH in the cocultures of mouse bone marrow cells with either SaOS-2/4 cells or SAOS-4/3 cells. Human OCLs were also formed in response to PTH in the coculture of SaOS-4/3 cells and human peripheral blood mononuclear cells. Adding dexamethasone together with PTH greatly enhanced PTH-induced human OCL formation. Like mouse OCLs, human OCLs formed in response to PTH were tartrate-resistant acid phosphatase positive, expressed abundant calcitonin receptors and vitronectin receptors, and formed resorption pits on dentine slices. Other osteotropic factors such as 1α,25-dihydroxyvitamin D3, prostaglandin E2, and interleukin 6 plus soluble interleukin 6 receptors failed to induce mouse and human OCLs in cocultures with SAOS-4/3 cells. Both mouse and human OCL formation supported by SAOS-4/3 cells were inhibited by either adding an antibody against macrophage-colony stimulating factor or adding granulocyte/macrophage-colony stimulating factor. Thus, it is likely that human and mouse OCL formation supported by SaOS-4/3 cells are similarly regulated. These results indicate that the target cells of PTH for inducing osteoclast formation are osteoblast/stromal cells but not osteoclast progenitor cells in the coculture. This coculture model will be useful for investigating the abnormalities of osteoclast differentiation and function in human metabolic bone diseases.",
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