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A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping

We recently described cell-projection pumping as a mechanism transferring cytoplasm between cells. The uptake of fibroblast cytoplasm by co-cultured SAOS-2 osteosarcoma cells changes SAOS-2 morphology and increases cell migration and proliferation, as seen by single-cell tracking and in FACS separat...

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Autores principales: Mahadevan, Swarna, Kwong, Kenelm, Lu, Mingjie, Kelly, Elizabeth, Chami, Belal, Romin, Yevgeniy, Fujisawa, Sho, Manova, Katia, Moore, Malcolm A. S., Zoellner, Hans
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9316567/
https://www.ncbi.nlm.nih.gov/pubmed/35887295
http://dx.doi.org/10.3390/ijms23147949
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author Mahadevan, Swarna
Kwong, Kenelm
Lu, Mingjie
Kelly, Elizabeth
Chami, Belal
Romin, Yevgeniy
Fujisawa, Sho
Manova, Katia
Moore, Malcolm A. S.
Zoellner, Hans
author_facet Mahadevan, Swarna
Kwong, Kenelm
Lu, Mingjie
Kelly, Elizabeth
Chami, Belal
Romin, Yevgeniy
Fujisawa, Sho
Manova, Katia
Moore, Malcolm A. S.
Zoellner, Hans
author_sort Mahadevan, Swarna
collection PubMed
description We recently described cell-projection pumping as a mechanism transferring cytoplasm between cells. The uptake of fibroblast cytoplasm by co-cultured SAOS-2 osteosarcoma cells changes SAOS-2 morphology and increases cell migration and proliferation, as seen by single-cell tracking and in FACS separated SAOS-2 from co-cultures. Morphological changes in SAOS-2 seen by single cell tracking are consistent with previous observations in fixed monolayers of SAOS-2 co-cultures. Notably, earlier studies with fixed co-cultures were limited by the absence of a quantitative method for identifying sub-populations of co-cultured cells, or for quantitating transfer relative to control populations of SAOS-2 or fibroblasts cultured alone. We now overcome that limitation by a novel Cartesian plot analysis that identifies individual co-cultured cells as belonging to one of five distinct cell populations, and also gives numerical measure of similarity to control cell populations. We verified the utility of the method by first confirming the previously established relationship between SAOS-2 morphology and uptake of fibroblast contents, and also demonstrated similar effects in other cancer cell lines including from melanomas, and cancers of the ovary and colon. The method was extended to examine global DNA methylation, and while there was no clear effect on SAOS-2 DNA methylation, co-cultured fibroblasts had greatly reduced DNA methylation, similar to cancer associated fibroblasts.
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spelling pubmed-93165672022-07-27 A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping Mahadevan, Swarna Kwong, Kenelm Lu, Mingjie Kelly, Elizabeth Chami, Belal Romin, Yevgeniy Fujisawa, Sho Manova, Katia Moore, Malcolm A. S. Zoellner, Hans Int J Mol Sci Article We recently described cell-projection pumping as a mechanism transferring cytoplasm between cells. The uptake of fibroblast cytoplasm by co-cultured SAOS-2 osteosarcoma cells changes SAOS-2 morphology and increases cell migration and proliferation, as seen by single-cell tracking and in FACS separated SAOS-2 from co-cultures. Morphological changes in SAOS-2 seen by single cell tracking are consistent with previous observations in fixed monolayers of SAOS-2 co-cultures. Notably, earlier studies with fixed co-cultures were limited by the absence of a quantitative method for identifying sub-populations of co-cultured cells, or for quantitating transfer relative to control populations of SAOS-2 or fibroblasts cultured alone. We now overcome that limitation by a novel Cartesian plot analysis that identifies individual co-cultured cells as belonging to one of five distinct cell populations, and also gives numerical measure of similarity to control cell populations. We verified the utility of the method by first confirming the previously established relationship between SAOS-2 morphology and uptake of fibroblast contents, and also demonstrated similar effects in other cancer cell lines including from melanomas, and cancers of the ovary and colon. The method was extended to examine global DNA methylation, and while there was no clear effect on SAOS-2 DNA methylation, co-cultured fibroblasts had greatly reduced DNA methylation, similar to cancer associated fibroblasts. MDPI 2022-07-19 /pmc/articles/PMC9316567/ /pubmed/35887295 http://dx.doi.org/10.3390/ijms23147949 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Mahadevan, Swarna
Kwong, Kenelm
Lu, Mingjie
Kelly, Elizabeth
Chami, Belal
Romin, Yevgeniy
Fujisawa, Sho
Manova, Katia
Moore, Malcolm A. S.
Zoellner, Hans
A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping
title A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping
title_full A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping
title_fullStr A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping
title_full_unstemmed A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping
title_short A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping
title_sort novel cartesian plot analysis for fixed monolayers that relates cell phenotype to transfer of contents between fibroblasts and cancer cells by cell-projection pumping
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9316567/
https://www.ncbi.nlm.nih.gov/pubmed/35887295
http://dx.doi.org/10.3390/ijms23147949
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