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Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies

The neuroblastoma cell line SH-SY5Y has been a well-established and very popular in vitro model in neuroscience for decades, especially focusing on neurodevelopmental disorders, such as Parkinson’s disease. The ability of this cell type to differentiate compared with other models in neurobiology mak...

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Autores principales: Feles, Sebastian, Overath, Christian, Reichardt, Sina, Diegeler, Sebastian, Schmitz, Claudia, Kronenberg, Jessica, Baumstark-Khan, Christa, Hemmersbach, Ruth, Hellweg, Christine E., Liemersdorf, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9326679/
https://www.ncbi.nlm.nih.gov/pubmed/35893584
http://dx.doi.org/10.3390/mps5040058
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author Feles, Sebastian
Overath, Christian
Reichardt, Sina
Diegeler, Sebastian
Schmitz, Claudia
Kronenberg, Jessica
Baumstark-Khan, Christa
Hemmersbach, Ruth
Hellweg, Christine E.
Liemersdorf, Christian
author_facet Feles, Sebastian
Overath, Christian
Reichardt, Sina
Diegeler, Sebastian
Schmitz, Claudia
Kronenberg, Jessica
Baumstark-Khan, Christa
Hemmersbach, Ruth
Hellweg, Christine E.
Liemersdorf, Christian
author_sort Feles, Sebastian
collection PubMed
description The neuroblastoma cell line SH-SY5Y has been a well-established and very popular in vitro model in neuroscience for decades, especially focusing on neurodevelopmental disorders, such as Parkinson’s disease. The ability of this cell type to differentiate compared with other models in neurobiology makes it one of the few suitable models without having to rely on a primary culture of neuronal cells. Over the years, various, partly contradictory, methods of cultivation have been reported. This study is intended to provide a comprehensive guide to the in vitro cultivation of undifferentiated SH-SY5Y cells. For this purpose, the morphology of the cell line and the differentiation of the individual subtypes are described, and instructions for cell culture practice and long-term cryoconservation are provided. We describe the key growth characteristics of this cell line, including proliferation and confluency data, optimal initial seeding cell numbers, and a comparison of different culture media and cell viability during cultivation. Furthermore, applying an optimized protocol in a long-term cultivation over 60 days, we show that cumulative population doubling (CPD) is constant over time and does not decrease with incremental passage, enabling stable cultivation, for example, for recurrent differentiation to achieve the highest possible reproducibility in subsequent analyses. Therefore, we provide a solid guidance for future research that employs the neuroblastoma cell line SH-SY5Y.
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spelling pubmed-93266792022-07-28 Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies Feles, Sebastian Overath, Christian Reichardt, Sina Diegeler, Sebastian Schmitz, Claudia Kronenberg, Jessica Baumstark-Khan, Christa Hemmersbach, Ruth Hellweg, Christine E. Liemersdorf, Christian Methods Protoc Article The neuroblastoma cell line SH-SY5Y has been a well-established and very popular in vitro model in neuroscience for decades, especially focusing on neurodevelopmental disorders, such as Parkinson’s disease. The ability of this cell type to differentiate compared with other models in neurobiology makes it one of the few suitable models without having to rely on a primary culture of neuronal cells. Over the years, various, partly contradictory, methods of cultivation have been reported. This study is intended to provide a comprehensive guide to the in vitro cultivation of undifferentiated SH-SY5Y cells. For this purpose, the morphology of the cell line and the differentiation of the individual subtypes are described, and instructions for cell culture practice and long-term cryoconservation are provided. We describe the key growth characteristics of this cell line, including proliferation and confluency data, optimal initial seeding cell numbers, and a comparison of different culture media and cell viability during cultivation. Furthermore, applying an optimized protocol in a long-term cultivation over 60 days, we show that cumulative population doubling (CPD) is constant over time and does not decrease with incremental passage, enabling stable cultivation, for example, for recurrent differentiation to achieve the highest possible reproducibility in subsequent analyses. Therefore, we provide a solid guidance for future research that employs the neuroblastoma cell line SH-SY5Y. MDPI 2022-07-12 /pmc/articles/PMC9326679/ /pubmed/35893584 http://dx.doi.org/10.3390/mps5040058 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
Feles, Sebastian
Overath, Christian
Reichardt, Sina
Diegeler, Sebastian
Schmitz, Claudia
Kronenberg, Jessica
Baumstark-Khan, Christa
Hemmersbach, Ruth
Hellweg, Christine E.
Liemersdorf, Christian
Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies
title Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies
title_full Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies
title_fullStr Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies
title_full_unstemmed Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies
title_short Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies
title_sort streamlining culture conditions for the neuroblastoma cell line sh-sy5y: a prerequisite for functional studies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9326679/
https://www.ncbi.nlm.nih.gov/pubmed/35893584
http://dx.doi.org/10.3390/mps5040058
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