Adenosine blocks aminopterin-induced suppression of osteoclast differentiation

Junpei Teramachi, Akiko Kukita, Pengfei Qu, Naohisa Wada, Yin Ji Li, Seiji Nakamura, Toshio Kukita

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

To search cell surface molecules involved in the regulation of osteoclastogenesis, especially in fusion process, it is one powerful approach to obtain monoclonal antibodies bearing ability to block formation of multinucleated osteoclasts. Ideally, direct bio-assay of hybridoma supernatants is quite convenient to screen monoclonal antibodies of interest from numerous culture wells. However, addition of hybridoma supernatant containing hypoxanthine-aminopterin-thymidine (HAT), components of the selection medium, to whole bone marrow cultures strikingly suppressed osteoclastogenesis. Here we clarified aminopterin is the responsible component in HAT medium to inhibit osteoclastogenesis. Methotrexate (MTX), mono-methylated aminopterin, showed similar suppressive effect on osteoclastogenesis. When bone marrow cells were cultured in the presence of all nucleosides, aminopterin and MTX-induced suppression of osteoclastogenesis was abrogated. Among four nucleosides only adenosine canceled aminopterin-induced suppression of osteoclastogenesis. Direct bio-assay of hybridoma supernatant containing HAT selection medium is now available to screen monoclonal antibodies if adenosine-containing culture medium was utilized for evaluating osteoclastogenesis.

Original languageEnglish
Pages (from-to)64-70
Number of pages7
JournalJournal of Bone and Mineral Metabolism
Volume31
Issue number1
DOIs
Publication statusPublished - Jan 1 2013

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Aminopterin
Osteoclasts
Osteogenesis
Adenosine
Hypoxanthine
Hybridomas
Thymidine
Monoclonal Antibodies
Nucleosides
Methotrexate
Bone Marrow Cells
Culture Media
Bone Marrow

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine
  • Endocrinology

Cite this

Adenosine blocks aminopterin-induced suppression of osteoclast differentiation. / Teramachi, Junpei; Kukita, Akiko; Qu, Pengfei; Wada, Naohisa; Li, Yin Ji; Nakamura, Seiji; Kukita, Toshio.

In: Journal of Bone and Mineral Metabolism, Vol. 31, No. 1, 01.01.2013, p. 64-70.

Research output: Contribution to journalArticle

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AU - Nakamura, Seiji

AU - Kukita, Toshio

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