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

Lothar Seefried, Sigrid Mueller-Deubert, Thomas Schwarz Wentzer, Thomas Lind, Birgit Mentrup, Melanie Kober, Denitsa Docheva, Astrid Liedert, Moustapha Kassem, Anita Ignatius, Matthias Schieker, Lutz Claes, Winfried Wilke, Franz Jakob, Regina Ebert

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissue-specific 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.
OriginalsprogEngelsk
TidsskriftEuropean Cells & Materials
Vol/bind20
Sider (fra-til)344-55
Antal sider12
ISSN1473-2262
StatusUdgivet - 1. jan. 2010

Fingeraftryk

Polyurethanes
Bioreactors
Reporter Genes
Cell culture
Signal transduction
Genes
Crosstalk
Luciferases
Stretching
Chemical activation
Tissue
Transcription factors
Osteoblasts
Stem cells
Screening
Bone
Transcription Factors
Phosphotransferases
Cells
Cellular Mechanotransduction

Citer dette

Seefried, L., Mueller-Deubert, S., Wentzer, T. S., Lind, T., Mentrup, B., Kober, M., ... Ebert, R. (2010). A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes. European Cells & Materials, 20, 344-55.
Seefried, Lothar ; Mueller-Deubert, Sigrid ; Wentzer, Thomas Schwarz ; Lind, Thomas ; Mentrup, Birgit ; Kober, Melanie ; Docheva, Denitsa ; Liedert, Astrid ; Kassem, Moustapha ; Ignatius, Anita ; Schieker, Matthias ; Claes, Lutz ; Wilke, Winfried ; Jakob, Franz ; Ebert, Regina. / A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes. I: European Cells & Materials. 2010 ; Bind 20. s. 344-55.
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title = "A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes",
abstract = "Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissue-specific 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.",
author = "Lothar Seefried and Sigrid Mueller-Deubert and Wentzer, {Thomas Schwarz} and Thomas Lind and Birgit Mentrup and Melanie Kober and Denitsa Docheva and Astrid Liedert and Moustapha Kassem and Anita Ignatius and Matthias Schieker and Lutz Claes and Winfried Wilke and Franz Jakob and Regina Ebert",
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Seefried, L, Mueller-Deubert, S, Wentzer, TS, Lind, T, Mentrup, B, Kober, M, Docheva, D, Liedert, A, Kassem, M, Ignatius, A, Schieker, M, Claes, L, Wilke, W, Jakob, F & Ebert, R 2010, 'A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes', European Cells & Materials, bind 20, s. 344-55.

A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes. / Seefried, Lothar; Mueller-Deubert, Sigrid; Wentzer, Thomas Schwarz; Lind, Thomas; Mentrup, Birgit; Kober, Melanie; Docheva, Denitsa; Liedert, Astrid; Kassem, Moustapha; Ignatius, Anita; Schieker, Matthias; Claes, Lutz; Wilke, Winfried; Jakob, Franz; Ebert, Regina.

I: European Cells & Materials, Bind 20, 01.01.2010, s. 344-55.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes

AU - Seefried, Lothar

AU - Mueller-Deubert, Sigrid

AU - Wentzer, Thomas Schwarz

AU - Lind, Thomas

AU - Mentrup, Birgit

AU - Kober, Melanie

AU - Docheva, Denitsa

AU - Liedert, Astrid

AU - Kassem, Moustapha

AU - Ignatius, Anita

AU - Schieker, Matthias

AU - Claes, Lutz

AU - Wilke, Winfried

AU - Jakob, Franz

AU - Ebert, Regina

PY - 2010/1/1

Y1 - 2010/1/1

N2 - Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissue-specific 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.

AB - Mechanical forces are translated into biochemical signals and contribute to cell differentiation and phenotype maintenance. Mesenchymal stem cells and their tissue-specific 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.

M3 - Journal article

C2 - 21154241

VL - 20

SP - 344

EP - 355

JO - European Cells & Materials

JF - European Cells & Materials

SN - 1473-2262

ER -

Seefried L, Mueller-Deubert S, Wentzer TS, Lind T, Mentrup B, Kober M et al. A small scale cell culture system to analyze mechanobiology using reporter gene constructs and polyurethane dishes. European Cells & Materials. 2010 jan 1;20:344-55.