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Bioengineering

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JoVE Journal Bioengineering
Isolation of Primary Human Colon Tumor Cells from Surgical Tissues and Culturing Them Directly on Soft Elastic Substrates for Traction Cytometry
 

Isolation of Primary Human Colon Tumor Cells from Surgical Tissues and Culturing Them Directly on Soft Elastic Substrates for Traction Cytometry

Article DOI: 10.3791/52532-v • 09:28 min • June 4th, 2015
June 4th, 2015 •
1, 1, 2,3, 2,3, 1,4
1Department of Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, 2College of Medicine, University of Illinois at Urbana-Champaign, 3Provena Covenant Medical Centre, 4Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign

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Summary
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A protocol is described to extract primary human cells from surgical colon tumor and normal tissues. The isolated cells are then cultured on soft elastic substrates (polyacrylamide hydrogels) functionalized by an extracellular matrix protein, and embedded with fluorescent microbeads. Traction cytometry is performed to assess cellular contractile stresses.

Tags

Isolation Primary Human Colon Tumor Cells Surgical Tissues Culturing Soft Elastic Substrates Traction Cytometry Cancer Cells Matrix Mechanical Stiffness Mechano-chemical System Adhesion Receptors Signal Transduction Membrane Proteins Cytoskeletal Architecture Molecular Motors Mechanosensitivity Immortalized Cell Lines Murine Derived Primary Cells Primary Human Cancer Cells Optimized Protocol Healthy And Cancerous Surgical Human Tissue Samples Soft Polyacrylamide Hydrogels Stiff Polyacrylamide Hydrogels Rigid Polystyrene Substrates Extracellular Matrix (fibronectin) Fluorescent Microbeads Traction Assay Cellular Contractile Stresses Immunofluorescence Microscopy Cell Morphology Cytoskeleton Organization Vinculin Containing Focal Adhesions
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