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A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between
Swarna Mahadevan1, Kenelm Kwong1, Mingjie Lu1
1The Cellular and Molecular Pathology Research Unit, Oral Pathology and Oral Medicine, School of Dentistry, Faculty of Medicine and Health, The University of Sydney, Westmead Hospital, Westmead, NSW 2145, Australia.
International Journal of Molecular Sciences
|July 27, 2022
Summary
Cell-projection pumping transfers cytoplasm between cells, altering cancer cell behavior and migration. A new Cartesian plot analysis quantifies this transfer and its effects on cell populations.
Area of Science:
- Cell Biology
- Cancer Research
- Biotechnology
Background:
- Cell-projection pumping facilitates cytoplasm transfer between cells.
- Previous studies observed morphological changes and increased migration/proliferation in SAOS-2 cells upon fibroblast cytoplasm uptake.
- Limitations existed in quantifying cell subpopulations and transfer rates in co-cultures.
Purpose of the Study:
- To develop a quantitative method for analyzing cell-projection pumping.
- To assess the impact of fibroblast cytoplasm uptake on various cancer cell lines.
- To investigate the effect of cell-projection pumping on DNA methylation.
Main Methods:
- Single-cell tracking and Fluorescence-Activated Cell Sorting (FACS) for cell separation.
- Novel Cartesian plot analysis to identify and quantify distinct cell populations in co-cultures.
- Global DNA methylation analysis.
Main Results:
- The Cartesian plot analysis successfully identified five distinct cell populations and quantified cytoplasm transfer.
- Fibroblast cytoplasm uptake altered SAOS-2 cell morphology and increased migration/proliferation, consistent with prior findings.
- Similar effects were observed in melanoma, ovarian, and colon cancer cell lines.
- Co-cultured fibroblasts exhibited significantly reduced DNA methylation, resembling cancer-associated fibroblasts, while SAOS-2 cells showed no clear methylation changes.
Conclusions:
- Cell-projection pumping is a significant mechanism for intercellular cytoplasm transfer with functional consequences for cancer cells.
- The developed Cartesian plot analysis provides a robust tool for quantitative assessment of cell-cell interactions and cytoplasm transfer.
- Fibroblast cytoplasm transfer impacts cancer cell behavior and can alter DNA methylation patterns in fibroblasts, suggesting potential implications for cancer biology.
