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Validation of a simple and fast method to quantify in vitro mineralization with fluorescent probes used in molecular imaging of bone
Authors:Martiene JC Moester  Monique AE Schoeman  Ineke B Oudshoorn  Mara M van Beusekom  Isabel M Mol  Eric L Kaijzel  Clemens WGM Löwik  Karien E de Rooij
Institution:1. Department of Radiology, Leiden University Medical Center, The Netherlands;2. Department of Orthopedic Surgery, Leiden University Medical Center, The Netherlands;3. Percuros BV, Leiden, The Netherlands
Abstract:Alizarin Red S staining is the standard method to indicate and quantify matrix mineralization during differentiation of osteoblast cultures. KS483 cells are multipotent mouse mesenchymal progenitor cells that can differentiate into chondrocytes, adipocytes and osteoblasts and are a well-characterized model for the study of bone formation. Matrix mineralization is the last step of differentiation of bone cells and is therefore a very important outcome measure in bone research. Fluorescently labelled calcium chelating agents, e.g. BoneTag and OsteoSense, are currently used for in vivo imaging of bone. The aim of the present study was to validate these probes for fast and simple detection and quantification of in vitro matrix mineralization by KS483 cells and thus enabling high-throughput screening experiments.
Keywords:Osteoblast  Mineralization  Fluorescence  Alizarin Red S  Staining  Quantification
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