Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation

Abstract Mammosphere assays are widely used in vitro to identify prospective cancer-initiating stem cells that can propagate clonally to form spheres in free-floating conditions. However, the traditional mammosphere assay inevitably introduces cell aggregation that interferes with the measurement of...

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Autores principales:	Lekhana Bhandary, Patrick C. Bailey, Katarina T. Chang, Karen F. Underwood, Cornell J. Lee, Rebecca A. Whipple, Christopher M. Jewell, Eleanor Ory, Keyata N. Thompson, Julia A. Ju, Trevor M. Mathias, Stephen J. P. Pratt, Michele I. Vitolo, Stuart S. Martin
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7fa9e6ad3962419dbe47d6107797d52b
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spelling	oai:doaj.org-article:7fa9e6ad3962419dbe47d6107797d52b2021-12-02T10:44:09ZLipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation10.1038/s41598-021-81919-92045-2322https://doaj.org/article/7fa9e6ad3962419dbe47d6107797d52b2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81919-9https://doaj.org/toc/2045-2322Abstract Mammosphere assays are widely used in vitro to identify prospective cancer-initiating stem cells that can propagate clonally to form spheres in free-floating conditions. However, the traditional mammosphere assay inevitably introduces cell aggregation that interferes with the measurement of true mammosphere forming efficiency. We developed a method to reduce tumor cell aggregation and increase the probability that the observed mammospheres formed are clonal in origin. Tethering individual tumor cells to lipid anchors prevents cell drift while maintaining free-floating characteristics. This enables real-time monitoring of single tumor cells as they divide to form mammospheres. Monitoring tethered breast cancer cells provided detailed size information that correlates directly to previously published single cell tracking data. We observed that 71% of the Day 7 spheres in lipid-coated wells were between 50 and 150 μm compared to only 37% in traditional low attachment plates. When an equal mixture of MCF7-GFP and MCF7-mCherry cells were seeded, 65% of the mammospheres in lipid-coated wells demonstrated single color expression whereas only 32% were single-colored in low attachment wells. These results indicate that using lipid tethering for mammosphere growth assays can reduce the confounding factor of cell aggregation and increase the formation of clonal mammospheres.Lekhana BhandaryPatrick C. BaileyKatarina T. ChangKaren F. UnderwoodCornell J. LeeRebecca A. WhippleChristopher M. JewellEleanor OryKeyata N. ThompsonJulia A. JuTrevor M. MathiasStephen J. P. PrattMichele I. VitoloStuart S. MartinNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lekhana Bhandary Patrick C. Bailey Katarina T. Chang Karen F. Underwood Cornell J. Lee Rebecca A. Whipple Christopher M. Jewell Eleanor Ory Keyata N. Thompson Julia A. Ju Trevor M. Mathias Stephen J. P. Pratt Michele I. Vitolo Stuart S. Martin Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
description	Abstract Mammosphere assays are widely used in vitro to identify prospective cancer-initiating stem cells that can propagate clonally to form spheres in free-floating conditions. However, the traditional mammosphere assay inevitably introduces cell aggregation that interferes with the measurement of true mammosphere forming efficiency. We developed a method to reduce tumor cell aggregation and increase the probability that the observed mammospheres formed are clonal in origin. Tethering individual tumor cells to lipid anchors prevents cell drift while maintaining free-floating characteristics. This enables real-time monitoring of single tumor cells as they divide to form mammospheres. Monitoring tethered breast cancer cells provided detailed size information that correlates directly to previously published single cell tracking data. We observed that 71% of the Day 7 spheres in lipid-coated wells were between 50 and 150 μm compared to only 37% in traditional low attachment plates. When an equal mixture of MCF7-GFP and MCF7-mCherry cells were seeded, 65% of the mammospheres in lipid-coated wells demonstrated single color expression whereas only 32% were single-colored in low attachment wells. These results indicate that using lipid tethering for mammosphere growth assays can reduce the confounding factor of cell aggregation and increase the formation of clonal mammospheres.
format	article
author	Lekhana Bhandary Patrick C. Bailey Katarina T. Chang Karen F. Underwood Cornell J. Lee Rebecca A. Whipple Christopher M. Jewell Eleanor Ory Keyata N. Thompson Julia A. Ju Trevor M. Mathias Stephen J. P. Pratt Michele I. Vitolo Stuart S. Martin
author_facet	Lekhana Bhandary Patrick C. Bailey Katarina T. Chang Karen F. Underwood Cornell J. Lee Rebecca A. Whipple Christopher M. Jewell Eleanor Ory Keyata N. Thompson Julia A. Ju Trevor M. Mathias Stephen J. P. Pratt Michele I. Vitolo Stuart S. Martin
author_sort	Lekhana Bhandary
title	Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
title_short	Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
title_full	Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
title_fullStr	Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
title_full_unstemmed	Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
title_sort	lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/7fa9e6ad3962419dbe47d6107797d52b
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Lipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation

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