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Automated Platform for Cell Selection and Separation Based on Four-Dimensional Motility and Matrix Degradation

preprint
submitted on 31.10.2019 and posted on 06.11.2019 by Hannah Nowotarski, Pete Attayek, Nancy Allbritton

Motility and invasion are key steps in the metastatic cascade, enabling cells to move through normal tissue borders into the surrounding stroma. Most available in vitro assays track cell motility or cell invasion but lack the ability to measure both simultaneously and then separate single cells with unique behaviors. In this work, we developed a cell-separation platform capable of tracking cell movement and invasion through an extracellular matrix in space and time. The platform utilized a collagen scaffold with embedded tumor cells overlaid onto a microraft array. Confocal microscopy enabled high resolution (0.4×0.4×3.5 µm voxel) monitoring of cell movement within the scaffolds. Two pancreatic cancer cell lines with known differing invasiveness were characterized on this platform, with median motilities of 14±6 mm and 10±4 mm over 48 h. Within the same cell line, cells demonstrated highly variable motility, with a range of XYZ movement from 144 mm to 2 mm over 24 h. The ten lowest and highest motility cells, with median movements of 33±11 mm and 3±1 mm, respectively, were separated and sub-cultured. After 6 weeks of culture, the cell populations were assayed on a Transwell invasion assay and 227±56 cells were invasive in the high motility population while only 48±10 cells were invasive in the low motility population, indicating that the resulting offspring possessed a motility phenotype reflective of the parental cells. This work demonstrates the feasibility of sorting single cells based on complex phenotypes along with the capability to further probe those cells and explore biological phenomena.

Funding

Single-Cell Measurement of Lipid Signaling in Colorectal Cancer

National Cancer Institute

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PROFILING SIGNALING ACTIVITY AND GENE EXPRESSION IN SINGLE, PANCREATIC ADENOCARCINOMA CELLS USING CE-RNA-SEQ

National Cancer Institute

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History

Email Address of Submitting Author

nowo@live.unc.edu

Institution

University of North Carolina at Chapel Hill

Country

United States of America

ORCID For Submitting Author

0000-0002-8702-7817

Declaration of Conflict of Interest

The authors declare the following competing financial interests: N.L.A. has a financial interest in Cell Microsystems, Inc.

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