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dc.contributor.authorBhandary, Lekhana
dc.contributor.authorBailey, Patrick C
dc.contributor.authorChang, Katarina T
dc.contributor.authorUnderwood, Karen F
dc.contributor.authorLee, Cornell J
dc.contributor.authorWhipple, Rebecca A
dc.contributor.authorJewell, Christopher M
dc.contributor.authorOry, Eleanor
dc.contributor.authorThompson, Keyata N
dc.contributor.authorJu, Julia A
dc.contributor.authorMathias, Trevor M
dc.contributor.authorPratt, Stephen J P
dc.contributor.authorVitolo, Michele I
dc.contributor.authorMartin, Stuart S
dc.date.accessioned2021-02-25T21:31:28Z
dc.date.available2021-02-25T21:31:28Z
dc.date.issued2021-02-05
dc.identifier.urihttp://hdl.handle.net/10713/14763
dc.description.abstractMammosphere 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. © 2021, The Author(s).en_US
dc.description.urihttps://doi.org/10.1038/s41598-021-81919-9en_US
dc.language.isoenen_US
dc.publisherSpringer Natureen_US
dc.relation.ispartofScientific Reportsen_US
dc.subjectmammosphereen_US
dc.subject.lcshBreast--Canceren_US
dc.subject.meshBreast Neoplasmsen_US
dc.subject.meshLipidsen_US
dc.subject.meshNeoplastic Stem Cellsen_US
dc.subject.meshClone Cellsen_US
dc.subject.meshCell Trackingen_US
dc.subject.meshMCF-7 Cellsen_US
dc.titleLipid tethering of breast tumor cells reduces cell aggregation during mammosphere formation.en_US
dc.typeArticleen_US
dc.identifier.doi10.1038/s41598-021-81919-9
dc.identifier.pmid33547369
dc.source.journaltitleScientific reports
dc.source.volume11
dc.source.issue1
dc.source.beginpage3214
dc.source.endpage
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryUnited States
dc.source.countryEngland


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