Characterization of 3D Printed Stretching Devices for Imaging Force Transmission in Live-Cells

Introduction Cell stretch is a method which can rapidly apply mechanical force through cell-matrix and cell-cell adhesions and can be utilized to better understand underlying biophysical questions related to intracellular force transmission and mechanotransduction. Methods 3D printable stretching de...

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Bibliographic Details
Published in:Cellular and molecular bioengineering 2019-08, Vol.12 (4), p.289-300
Main Authors: Mayer, Carl R., Arsenovic, Paul T., Bathula, Kranthidhar, Denis, Kevin B., Conway, Daniel E.
Format: Article
Language:English
Subjects:
Actin
Biological and Medical Physics
Biomaterials
Biomedical Engineering and Bioengineering
Biomedical Engineering/Biotechnology
Biophysics
Biosensors
Cell Biology
Cytology
Cytoskeleton
Deformation
Devices
Engineering
Finite element method
Fluorescence resonance energy transfer
Fluoroscopic imaging
Mechanical unloading
Mechanotransduction
Morphology
Nuclei (cytology)
Plastic deformation
Questions
Stretching
Three dimensional printing
Unloading
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Summary:Introduction Cell stretch is a method which can rapidly apply mechanical force through cell-matrix and cell-cell adhesions and can be utilized to better understand underlying biophysical questions related to intracellular force transmission and mechanotransduction. Methods 3D printable stretching devices suitable for live-cell fluorescent imaging were designed using finite element modeling and validated experimentally. These devices were then used along with FRET based nesprin-2G force sensitive biosensors as well as live cell fluorescent staining to understand how the nucleus responds to externally applied mechanical force in cells with both intact LINC (linker of nucleoskeleton and cytoskeleton) complex and cells with the LINC complex disrupted using expression of dominant negative KASH protein. Results The devices were shown to provide a larger strain ranges (300% uniaxial and 60% biaxial) than currently available commercial or academic designs we are aware of. Under uniaxial deformation, the deformation of the nucleus of NIH 3T3 cells per unit of imposed cell strain was shown to be approximately 50% higher in control cells compared to cells with a disrupted LINC complex. Under biaxial deformation, MDCK II cells showed permanent changes in the nuclear morphology as well as actin organization upon unloading, indicating that failure, plastic deformation, or remodeling of the cytoskeleton is occurring in response to the applied stretch. Conclusion Development and open distribution of low-cost, 3D-printable uniaxial and biaxial cell stretching devices compatible with live-cell fluorescent imaging allows a wider range of researchers to investigate mechanical influences on biological questions with only a minimal investment of resources.
ISSN:1865-5025
1865-5033
DOI:10.1007/s12195-019-00579-y