Development of high-throughput screening system for osteogenic drugs using a cell-based sensor

Hironori Hojo, Kazuyo Igawa, Shinsuke Ohba, Fumiko Yano, Keiji Nakajima, Yuske Komiyama, Toshiyuki Ikeda, Alexander C. Lichtler, Je Tae Woo, Takayuki Yonezawa, Tsuyoshi Takato, Ung il Chung

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

To effectively treat osteoporosis and other bone-loss disorders, small compounds that potently induce bone formation are needed. The present study initially attempted to establish a monitoring system that could detect osteogenic differentiation easily, precisely, and noninvasively. For this purpose, we established pre-osteoblastic MC3T3E1 cells stably transfected with the GFP reporter gene driven by a 2.3 kb fragment of rat type I collagen promoter (Col1a1GFP-MC3T3E1). Among these cells, we selected a clone that fluoresced upon osteogenic stimulation by BMP2. The GFP fluorescence intensity corresponded well to the intensity of alkaline phosphatase (ALP) staining and to the level of osteocalcin (Oc) mRNA. Using this system, we screened natural and synthetic compound libraries and thus identified an isoflavone derivative, glabrisoflavone (GI). GI induced ALP staining and Oc mRNA in a dose-dependent manner. The Col1a1GFP-MC3T3E1 system may be useful for identifying novel osteogenic drugs.

Original languageEnglish
Pages (from-to)375-379
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume376
Issue number2
DOIs
Publication statusPublished - Nov 14 2008
Externally publishedYes

Fingerprint

Osteocalcin
Alkaline Phosphatase
Screening
Bone
Throughput
Staining and Labeling
Messenger RNA
Isoflavones
Sensors
Collagen Type I
Reporter Genes
Osteogenesis
Pharmaceutical Preparations
Osteoporosis
Libraries
Rats
Clone Cells
Genes
Fluorescence
Bone and Bones

Keywords

  • Col1a1GFP-MC3T3E1
  • Osteoblast
  • Screening system
  • Small compounds

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Development of high-throughput screening system for osteogenic drugs using a cell-based sensor. / Hojo, Hironori; Igawa, Kazuyo; Ohba, Shinsuke; Yano, Fumiko; Nakajima, Keiji; Komiyama, Yuske; Ikeda, Toshiyuki; Lichtler, Alexander C.; Woo, Je Tae; Yonezawa, Takayuki; Takato, Tsuyoshi; Chung, Ung il.

In: Biochemical and Biophysical Research Communications, Vol. 376, No. 2, 14.11.2008, p. 375-379.

Research output: Contribution to journalArticle

Hojo, H, Igawa, K, Ohba, S, Yano, F, Nakajima, K, Komiyama, Y, Ikeda, T, Lichtler, AC, Woo, JT, Yonezawa, T, Takato, T & Chung, UI 2008, 'Development of high-throughput screening system for osteogenic drugs using a cell-based sensor', Biochemical and Biophysical Research Communications, vol. 376, no. 2, pp. 375-379. https://doi.org/10.1016/j.bbrc.2008.08.167
Hojo, Hironori ; Igawa, Kazuyo ; Ohba, Shinsuke ; Yano, Fumiko ; Nakajima, Keiji ; Komiyama, Yuske ; Ikeda, Toshiyuki ; Lichtler, Alexander C. ; Woo, Je Tae ; Yonezawa, Takayuki ; Takato, Tsuyoshi ; Chung, Ung il. / Development of high-throughput screening system for osteogenic drugs using a cell-based sensor. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 376, No. 2. pp. 375-379.
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