A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

Eric B. Berens; Jon M. Holy; Anna T. Riegel; Anton Wellstein

doi:10.3791/53409

A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

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Medicine

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JoVE Journal Medicine

A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting

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05:34 min

November 20, 2015

DOI: 10.3791/53409-v

Eric B. Berens¹, Jon M. Holy², Anna T. Riegel¹, Anton Wellstein¹

¹Lombardi Comprehensive Cancer Center, Department of Oncology,Georgetown University, ²Department of Biomedical Sciences,University of Minnesota

Overview

This method evaluates cancer cell invasion from spheroids into a surrounding 3D matrix. Spheroids are generated via the hanging drop culture method and then embedded in a matrix comprised of basement membrane materials and type I collagen. Invasion out of the spheroids is subsequently monitored.

Key Study Components

Area of Science

Cancer Biology
Cell Invasion
3D Cell Culture

Background

This method allows for monitoring cancer cell invasion from a cellular bolus.
It helps identify conditions that promote or inhibit cell invasion.
The technique evaluates cell invasion under physiologically relevant settings in vitro.
Utilizes a 3D matrix for realistic modeling of cancer cell behavior.

Purpose of Study

To assess cancer cell invasion dynamics.
To explore factors influencing cell invasion.
To provide insights into cancer biology mechanisms.

Methods Used

Remove a cultured dish with adherent cancer cells from the incubator.
Wash cells with PBS and add 1.5 milliliters of 0.05% trypsin EDTA.
Incubate cells for three minutes at 37 degrees Celsius.
Embed spheroids in a matrix of basement membrane materials and type I collagen.

Main Results

Successful monitoring of cancer cell invasion from spheroids.
Evaluation of invasion under controlled conditions.
Insights into the mechanisms of cancer cell behavior.
Identification of potential therapeutic targets.

Conclusions

The method provides a reliable way to study cancer cell invasion.
It enhances understanding of cancer biology.
Future studies can build on this technique for therapeutic development.

Frequently Asked Questions

What is the significance of using a 3D matrix?

A 3D matrix provides a more physiologically relevant environment for studying cell behavior compared to traditional 2D cultures.

How are spheroids generated in this method?

Spheroids are generated using the hanging drop culture method, which allows for the formation of compact cellular aggregates.

What conditions can be tested using this assay?

The assay can be used to test various conditions that may promote or inhibit cancer cell invasion.

What are the advantages of this technique?

The technique allows for detailed evaluation of cell invasion dynamics in a controlled, relevant environment.

Can this method be applied to other types of cells?

Yes, while this method focuses on cancer cells, it can potentially be adapted for other cell types as well.

What is the role of type I collagen in the matrix?

Type I collagen provides structural support and mimics the extracellular matrix found in tissues, facilitating realistic cell behavior.

The overall goal of this cancer cell steroid invasion assay is to allow for the monitoring of cancer cell invasion out of a cellular bolus and into a surrounding three dimensional matrix. This method can help answer key questions in cancer biology, like the identification of conditions that promote or inhibit cell invasion. The main advantage of this technique is that the various nuances of how cells invade can be evaluated under a more physiologically relevant setting in vitro.

To begin, remove a cultured dish containing adherent cancer cells from the incubator. Wash the cells once with PBS and add 1.5 milliliters of 0.05%tripsin EDTA. Then incubate the cells for three minutes at 37 degrees Celsius.

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