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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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Isolation of Primary Human Colon Tumor Cells from Surgical Tissues and Culturing Them Directly on Soft Elastic Substrates for Traction Cytometry
Bölümler
- 00:05Başlık
- 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
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.
Etiketler
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
