Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

Abstract Stroke causes death and disability globally but no neuroprotectant is approved for post-stroke neuronal injury. Neuroprotective compounds can be identified using oxygen glucose deprivation (OGD) of neuronal cells as an in vitro stroke model. Nerve growth factor (NGF)-differentiated PC12 phe...

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Autores principales:	PinFen Chua, William K. Lim
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/704ee4132a274bdaa404810324e826c4
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spelling	oai:doaj.org-article:704ee4132a274bdaa404810324e826c42021-12-02T18:03:31ZOptimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents10.1038/s41598-021-87431-42045-2322https://doaj.org/article/704ee4132a274bdaa404810324e826c42021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-87431-4https://doaj.org/toc/2045-2322Abstract Stroke causes death and disability globally but no neuroprotectant is approved for post-stroke neuronal injury. Neuroprotective compounds can be identified using oxygen glucose deprivation (OGD) of neuronal cells as an in vitro stroke model. Nerve growth factor (NGF)-differentiated PC12 pheochromocytoma cells are frequently used. However, investigators often find their clonal variant undifferentiable and are uncertain of optimal culture conditions. Hence we studied 3 commonly used PC12 variants: PC12 Adh, PC12 from Riken Cell Bank (PC12 Riken) and Neuroscreen-1 (NS-1) cells. We found DMEM the optimal media for PC12 Riken and NS-1 cells. Using a novel serum-free media approach, we identified collagen IV as the preferred adhesive substrate for both cell lines. We found PC12 Adh cells cannot attach without serum and is unable to differentiate using NGF. NS-1 cells differentiated to a maximal 72.7 ± 5.2% %, with substantial basal differentiation. We optimised differentiated NS-1 cells for an in vitro stroke model using 3 h of OGD resulting in ~ 70% viable cells. We screened 5 reported neuroprotectants and provide the first report that serotonin is antiapoptotic in a stroke model and the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) is neuroprotective in PC12 cells. Thus we demonstrate the optimisation and validation for a PC12 cell-based in vitro stroke model.PinFen ChuaWilliam K. LimNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q PinFen Chua William K. Lim Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents
description	Abstract Stroke causes death and disability globally but no neuroprotectant is approved for post-stroke neuronal injury. Neuroprotective compounds can be identified using oxygen glucose deprivation (OGD) of neuronal cells as an in vitro stroke model. Nerve growth factor (NGF)-differentiated PC12 pheochromocytoma cells are frequently used. However, investigators often find their clonal variant undifferentiable and are uncertain of optimal culture conditions. Hence we studied 3 commonly used PC12 variants: PC12 Adh, PC12 from Riken Cell Bank (PC12 Riken) and Neuroscreen-1 (NS-1) cells. We found DMEM the optimal media for PC12 Riken and NS-1 cells. Using a novel serum-free media approach, we identified collagen IV as the preferred adhesive substrate for both cell lines. We found PC12 Adh cells cannot attach without serum and is unable to differentiate using NGF. NS-1 cells differentiated to a maximal 72.7 ± 5.2% %, with substantial basal differentiation. We optimised differentiated NS-1 cells for an in vitro stroke model using 3 h of OGD resulting in ~ 70% viable cells. We screened 5 reported neuroprotectants and provide the first report that serotonin is antiapoptotic in a stroke model and the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) is neuroprotective in PC12 cells. Thus we demonstrate the optimisation and validation for a PC12 cell-based in vitro stroke model.
format	article
author	PinFen Chua William K. Lim
author_facet	PinFen Chua William K. Lim
author_sort	PinFen Chua
title	Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents
title_short	Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents
title_full	Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents
title_fullStr	Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents
title_full_unstemmed	Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents
title_sort	optimisation of a pc12 cell-based in vitro stroke model for screening neuroprotective agents
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/704ee4132a274bdaa404810324e826c4
work_keys_str_mv	AT pinfenchua optimisationofapc12cellbasedinvitrostrokemodelforscreeningneuroprotectiveagents AT williamklim optimisationofapc12cellbasedinvitrostrokemodelforscreeningneuroprotectiveagents
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Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

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