Automated platform for cell selection and separation based on four-dimensional motility and matrix degradation

Area of Science:

• Biomedical Engineering
• Cancer Biology
• Cellular Mechanobiology

Background:

• Metastasis involves cell motility and invasion, crucial steps in cancer progression.
• Existing in vitro assays often fail to simultaneously measure and separate cells based on these complex behaviors.
• Understanding single-cell heterogeneity in motility and invasion is vital for cancer research.

Purpose of the Study:

• To develop and validate a novel cell-separation platform for simultaneous tracking of cell movement (chemokinesis) and invasion.
• To isolate single cancer cells exhibiting distinct motility phenotypes.
• To investigate whether isolated single cells pass on their motility phenotype to their offspring.

Main Methods:

• Development of a cell-separation platform using a collagen scaffold with embedded tumor cells on a microraft array.
• High-resolution confocal microscopy for real-time monitoring of cell movement within the scaffold.
• Separation and sub-culturing of single cells with the highest and lowest motility.
• Assessment of the invasive potential of offspring populations using a Transwell invasion assay.

Main Results:

• The platform successfully monitored cell movement and invasion in space and time with high resolution.
• Significant variability in motility was observed within the same cancer cell line.
• Isolated high-motility cells produced offspring with significantly higher invasion rates (227 ± 56 cells) compared to offspring from low-motility cells (48 ± 10 cells).
• The motility phenotype was demonstrated to be heritable.

Conclusions:

• The developed platform enables simultaneous tracking and separation of single cells based on complex motility and invasion phenotypes.
• This technology allows for the isolation of distinct cell populations with heritable phenotypic traits.
• This approach offers a powerful tool for dissecting cellular heterogeneity and its role in cancer metastasis.