Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/15178
Title: Implementation of a Drug Discovery Tool for the Evaluation of Anti-Fibrotic Compounds: Application in Fibrovascular Disorders
Authors: IRMA ARSIANTI
Keywords: collagen, macromolecular crowding, dextran sulfate, collagen quantification, cell enumeration, drug discovery tool
Issue Date: 22-May-2006
Source: IRMA ARSIANTI (2006-05-22). Implementation of a Drug Discovery Tool for the Evaluation of Anti-Fibrotic Compounds: Application in Fibrovascular Disorders. ScholarBank@NUS Repository.
Abstract: In this project, the principle of macromolecular crowding was applied in a fibroblast culture system to enhance the formation of collagen matrix. We have successfully demonstrated that dextran sulfate (DexS), a polyanionic macromolecule, creates a volume exclusion effect in the culture medium, and thus accelerates the enzymatic processing of procollagen to collagen, and its subsequent deposition.Gel electrophoresis and Western blotting revealed that in normal fibroblast culture, most procollagen remained unconverted in the culture medium. The addition of DexS resulted in the association of collagen with the cell layer. This observation was confirmed with immunocytochemistry.A fluorometric-based anti-fibrotic drug discovery tool was also developed in this project. This tool integrated the cell enumeration assay and the collagen quantification assay on one plate. The cell enumeration assay was based on the measurement of the DAPI-stained nucleus, whereas the collagen quantification assay was based on an immunocytochemistry technique.
URI: http://scholarbank.nus.edu.sg/handle/10635/15178
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