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

DOI:
10.3791/52532-v

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09:28 min

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June 04, 2015

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, , , ,
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

Chapters

  • 00:05Title
  • 01:25Collection and Digestion of Surgical Tissue Sample
  • 02:40Dissociation of Tissue Sample into Single Cell Suspension and Culturing Isolated Cells on Substrates with Different Rigidity
  • 04:45Traction Force Microscopy
  • 05:43Immunofluorescence Microscopy Assays
  • 07:10Results: Traction Cytometry and Immunofluorescence Microscopy
  • 08:51Conclusion

Summary

Automatic Translation

Automatic Translation

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

Keywords: Primary Human Colon Tumor CellsSoft Elastic SubstratesTraction CytometryMatrix Mechanical StiffnessAdhesion ReceptorsSignal TransductionCytoskeletonMolecular MotorsMechanosensitivityImmortalized Cell LinesMurine Derived Primary CellsPolyacrylamide HydrogelsExtracellular MatrixFibronectinTraction AssayCellular Contractile StressesImmunofluorescence MicroscopyCell MorphologyCytoskeleton OrganizationVinculinFocal Adhesions

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