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Overview
This video demonstrates a fluorescence-based methodology to measure cell invasion through cell culture inserts. The procedure highlights the importance of cell invasion in cancer progression and angiogenesis.
Key Study Components
Area of Science
- Cell biology
- Cancer research
- In vitro assays
Background
- Cell invasion is critical in cancer progression.
- In vitro assays assess the ability of cells to migrate through barriers.
- Fluorescence-based methods allow for the detection of invasive cells.
- Different cell lines can serve as positive and negative controls.
Purpose of Study
- To demonstrate a procedure for measuring tumor cell invasion.
- To provide a method for studying the interaction of cells with a basement membrane.
- To compare invasive and non-invasive cell lines.
Methods Used
- Rehydration of the BD bio coat tumor invasion system.
- Cell preparation and seeding in culture inserts.
- Incubation with chemo attractants to promote cell migration.
- Fluorescent staining and measurement of cell invasion.
Main Results
- HT 10 80 cells showed significant invasion through the matrigel.
- NIH 3T3 cells did not invade but migrated through the uncoated membrane.
- Fluorescence measurements correlated with visual observations.
- Percent invasion was calculated based on fluorescent readings.
Conclusions
- The procedure effectively demonstrates tumor cell invasion.
- Fluorescence-based assays provide reliable data on cell behavior.
- Cell lines can be utilized to study invasive properties in cancer research.
What is the significance of cell invasion in cancer?
Cell invasion is a key step in cancer metastasis, allowing tumor cells to spread to other parts of the body.
How are the cells prepared for the invasion assay?
Cells are trypsinized, resuspended in serum-free medium, and seeded into the culture inserts.
What controls are used in the assay?
HT 10 80 cells serve as positive controls, while NIH 3T3 cells act as negative controls.
How is cell invasion measured?
Invasion is measured using fluorescence intensity after staining the cells that have migrated through the matrigel.
What are the incubation conditions for the assay?
The assay is incubated at 37 degrees Celsius with 5% carbon dioxide for optimal cell growth.
Can different cell types be used in this assay?
Yes, different cell types can be tested, but optimal seeding densities must be determined empirically.
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