Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

Area of Science:

• Cell Biology
• Biotechnology
• Cancer Research

Background:

• Traditional 3D invasion assays disrupt cellular microenvironments and cause cell loss.
• Studying rare or sensitive cells in their native microenvironment is challenging.
• Existing methods necessitate cell displacement, altering intrinsic cellular processes.

Purpose of the Study:

• To develop a novel 3D invasion assay that maintains microenvironmental integrity.
• To enable the study of rare and sensitive cells without displacement.
• To characterize the invasive capacity of hybrid cells formed from breast cancer cells and stem cells.

Main Methods:

• Cells are plated in a well, and an extracellular matrix (ECM) with chemoattractant is overlaid.
• The assay allows cells to invade upwards into the ECM matrix.
• Cell invasion and morphology are assessed within the collagen gel matrix.

Main Results:

• The new assay avoids cell displacement and preserves the microenvironment.
• It successfully enables the assessment of cell invasion and morphology.
• The invasive capacity of rare hybrid cells (MCF7-MSC fusions) was characterized.

Conclusions:

• The developed 3D invasion assay is suitable for studying rare and sensitive cells.
• This method preserves cellular microenvironment integrity during invasion studies.
• It provides a valuable tool for cancer research, particularly for investigating cell fusion and invasion.