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

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• 作者：Hironori Hojo ; Kazuyo Igawa ; Shinsuke Ohba ; Fumiko Yano ; Keiji Nakajima ; Yuske Komiyama ; Toshiyuki Ikeda ; Alex ; er C Lichtler ; Je-Tae Woo ; Takayuki Yonezawa ; Tsuyoshi Takato ; Ung-il Chung
• 关键词：Screening system ; Col1a1GFP-MC3T3E1 ; Small compounds ; Osteoblast
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2008
• 出版时间：14 November 2008
• 年：2008
• 卷：376
• 期：2
• 页码：375-379
• 全文大小：504 K

文摘

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.