Patient derived glioma stem cell spheroid reporter assays for live cell high content analysis

Jayne Culley; Peter W Nagle; John C Dawson; Neil O Carragher

doi:10.1016/j.slasd.2022.09.002

Patient derived glioma stem cell spheroid reporter assays for live cell high content analysis

Jayne Culley, Peter W Nagle, John C Dawson, Neil O Carragher^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Three dimensional models of cell culture enables researchers to recreate aspects of tumour biology not replicated by traditional two dimensional techniques. Here we describe a protocol to enable automated high throughput phenotypic profiling across panels of patient derived glioma stem cell spheroid models. We demonstrate the use of both live/dead cell end-points and monitor the dynamic changes in the cell cycle using cell lines expressing the FUCCI cell cycle reporter. Together, these assays provide additional insight into the mechanism of action of compound treatments over traditional cell viability assay endpoints.

Original language	English
Journal	Slas Discovery
Early online date	15 Sep 2022
DOIs	https://doi.org/10.1016/j.slasd.2022.09.002
Publication status	E-pub ahead of print - 15 Sep 2022

Keywords

Glioblastoma
3-dimensional
Spheroid
Cell Cycle
Drug Discovery

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10.1016/j.slasd.2022.09.002Licence: CC BY-NC-ND

Patient derived glioma stem cell spheroid reporter assays for live cell high content analysisFinal published version, 3 MBLicence: CC BY-NC-ND

https://linkinghub.elsevier.com/retrieve/pii/S2472555222136944

1 Active

Systems approach to therapeutic combinations for glioblastoma: new targets and agents delivered and sequenced for synergy
Carragher, N. & Frame, M.
UK-based charities
1/09/19 → 31/08/24
Project: Research

Cite this

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title = "Patient derived glioma stem cell spheroid reporter assays for live cell high content analysis",

abstract = "Three dimensional models of cell culture enables researchers to recreate aspects of tumour biology not replicated by traditional two dimensional techniques. Here we describe a protocol to enable automated high throughput phenotypic profiling across panels of patient derived glioma stem cell spheroid models. We demonstrate the use of both live/dead cell end-points and monitor the dynamic changes in the cell cycle using cell lines expressing the FUCCI cell cycle reporter. Together, these assays provide additional insight into the mechanism of action of compound treatments over traditional cell viability assay endpoints.",

keywords = "Glioblastoma, 3-dimensional, Spheroid, Cell Cycle, Drug Discovery",

author = "Jayne Culley and Nagle, {Peter W} and Dawson, {John C} and Carragher, {Neil O}",

note = "Funding Information: This work was supported by a Cancer Research UK Accelerator award ( A29368 ) and a joint Cancer Research UK ( A28596 ) and The Brain Tumour Charity award ( GN-000676 ) to N.O.C. We also acknowledge Professor Steven Pollard and Dr Gillian Morrison and the CRUK Glioma Cellular Genetic resource for the provision of patient derived GSCs. Funding Information: Neil Carragher reports financial support was provided by Cancer Research UK. Neil Carragher reports financial support was provided by Brain Tumour Charity. Neil Carragher reports a relationship with Cancer Research UK that includes: funding grants and travel reimbursement. Publisher Copyright: {\textcopyright} 2022",

year = "2022",

month = sep,

day = "15",

doi = "10.1016/j.slasd.2022.09.002",

language = "English",

journal = "Slas Discovery",

issn = "2472-5552",

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T1 - Patient derived glioma stem cell spheroid reporter assays for live cell high content analysis

AU - Culley, Jayne

AU - Nagle, Peter W

AU - Dawson, John C

AU - Carragher, Neil O

N1 - Funding Information: This work was supported by a Cancer Research UK Accelerator award ( A29368 ) and a joint Cancer Research UK ( A28596 ) and The Brain Tumour Charity award ( GN-000676 ) to N.O.C. We also acknowledge Professor Steven Pollard and Dr Gillian Morrison and the CRUK Glioma Cellular Genetic resource for the provision of patient derived GSCs. Funding Information: Neil Carragher reports financial support was provided by Cancer Research UK. Neil Carragher reports financial support was provided by Brain Tumour Charity. Neil Carragher reports a relationship with Cancer Research UK that includes: funding grants and travel reimbursement. Publisher Copyright: © 2022

PY - 2022/9/15

Y1 - 2022/9/15

N2 - Three dimensional models of cell culture enables researchers to recreate aspects of tumour biology not replicated by traditional two dimensional techniques. Here we describe a protocol to enable automated high throughput phenotypic profiling across panels of patient derived glioma stem cell spheroid models. We demonstrate the use of both live/dead cell end-points and monitor the dynamic changes in the cell cycle using cell lines expressing the FUCCI cell cycle reporter. Together, these assays provide additional insight into the mechanism of action of compound treatments over traditional cell viability assay endpoints.

AB - Three dimensional models of cell culture enables researchers to recreate aspects of tumour biology not replicated by traditional two dimensional techniques. Here we describe a protocol to enable automated high throughput phenotypic profiling across panels of patient derived glioma stem cell spheroid models. We demonstrate the use of both live/dead cell end-points and monitor the dynamic changes in the cell cycle using cell lines expressing the FUCCI cell cycle reporter. Together, these assays provide additional insight into the mechanism of action of compound treatments over traditional cell viability assay endpoints.

KW - Glioblastoma

KW - 3-dimensional

KW - Spheroid

KW - Cell Cycle

KW - Drug Discovery

U2 - 10.1016/j.slasd.2022.09.002

DO - 10.1016/j.slasd.2022.09.002

M3 - Article

JO - Slas Discovery

JF - Slas Discovery

SN - 2472-5552

ER -

Patient derived glioma stem cell spheroid reporter assays for live cell high content analysis

Abstract

Keywords

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Projects

Systems approach to therapeutic combinations for glioblastoma: new targets and agents delivered and sequenced for synergy

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