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A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes

Please always quote using this URN: urn:nbn:de:bvb:20-opus-68099
  • Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissuespecific offspring, as osteoblasts and chondrocytes, cells of cardiovascular tissues and lung cells are sensitive to mechanical loading but molecules and mechanisms involved have to be unraveled. It is well established that cellular mechanotransduction is mediated e.g. by activation of the transcription factor SP1 and by kinase signaling cascades resulting in the activation of the AP1Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissuespecific offspring, as osteoblasts and chondrocytes, cells of cardiovascular tissues and lung cells are sensitive to mechanical loading but molecules and mechanisms involved have to be unraveled. It is well established that cellular mechanotransduction is mediated e.g. by activation of the transcription factor SP1 and by kinase signaling cascades resulting in the activation of the AP1 complex. To investigate cellular mechanisms involved in mechanotransduction and to analyze substances, which modulate cellular mechanosensitivity reporter gene constructs, which can be transfected into cells of interest might be helpful. Suitable small-scale bioreactor systems and mechanosensitive reporter gene constructs are lacking. To analyze the molecular mechanisms of mechanotransduction and its crosstalk with biochemically induced signal transduction, AP1 and SP1 luciferase reporter gene constructs were cloned and transfected into various cell lines and primary cells. A newly developed bioreactor and small-scale 24-well polyurethane dishes were used to apply cyclic stretching to the transfected cells. 1 Hz cyclic stretching for 30 min in this system resulted in a significant stimulation of AP1 and SP1 mediated luciferase activity compared to unstimulated cells. In summary we describe a small-scale cell culture/bioreactor system capable of analyzing subcellular crosstalk mechanisms in mechanotransduction, mechanosensitivity of primary cells and of screening the activity of putative mechanosensitizers as new targets, e.g. for the treatment of bone loss caused by both disuse and signal transduction related alterations of mechanotransduction.show moreshow less

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Metadaten
Author: Lothar Seefried, Sigrid Mueller-Deubert, Thomas Schwarz, Thomas Lind, Birgit Mentrup, Melanie Kober, Denitsa Docheva, Astrid Liedert, Moustapha Kassem, Anita Ignatius, Matthias Schieker, Lutz Claes, Winfried Wilke, Franz Jakob, Regina Ebert
URN:urn:nbn:de:bvb:20-opus-68099
Document Type:Journal article
Faculties:Medizinische Fakultät / Lehrstuhl für Orthopädie
Language:English
Year of Completion:2010
Source:In: European Cells & Materials (2010) 20, 344-355.
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
GND Keyword:Bioreaktor
Tag:Mechanical strain; bioreactor; gene constructs; mechanosensitive reporter
Release Date:2012/10/09
Licence (German):License LogoDeutsches Urheberrecht