✓Establish 3D tumor spheroids and culture them in natural biomatrix
✓Quantify tumor cell invasion using semi-automated microscopy and image analysis
✓Interpret invasion assay results across multiple cancer model types
Protocol
Invasion of surrounding normal tissues is a defining characteristic of malignant tumors. We provide here a simple, semi-automated micro-plate assay of invasion into a natural 3D biomatrix that has been exemplified with a number of models of advanced human cancers.
Difficulty
intermediate
Total time
~5-7 days (spheroid generation and invasion assay)
Model organism
Human cancer cell lines (advanced tumor models)
Biosafety
BSL-2
Steps
1
Prepare and culture 3D tumor spheroids
Generate three-dimensional tumor spheroids from cancer cell lines using appropriate culture conditions. Allow spheroids to reach appropriate size and maturity before invasion assay initiation.
▶ 02:32
2
Embed spheroids in natural 3D biomatrix
Place mature tumor spheroids into wells containing natural extracellular matrix material. This creates a physiologically relevant microenvironment for invasion assessment.
▶ 04:57
3
Acquire images using automated microscopy system
Set up and execute automated image acquisition at defined timepoints to capture spheroid invasion into the biomatrix. Configure image analysis software for quantification.
▶ 07:21
4
Perform manual image acquisition and analysis
Manually image spheroids and measure invasion parameters when automated systems are unavailable. Quantify invaded cell area and distance using standard image analysis tools.
▶ 09:36
5
Compare invasion across cancer model types
Analyze and interpret invasion assay results across different human cancer cell lines and models. Demonstrate differential invasion behavior patterns.
▶ 11:22
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