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Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions

Metabolomic profiling of cell lines has shown many potential applications and advantages compared to animal models and human subjects, and an accurate cellular metabolite analysis is critical to understanding both the intracellular and extracellular environments in cell culture. This study provides...

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Autores principales: Campanella, Beatrice, Colombaioni, Laura, Nieri, Riccardo, Benedetti, Edoardo, Onor, Massimo, Bramanti, Emilia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8085660/
https://www.ncbi.nlm.nih.gov/pubmed/33937186
http://dx.doi.org/10.3389/fchem.2021.621548
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author Campanella, Beatrice
Colombaioni, Laura
Nieri, Riccardo
Benedetti, Edoardo
Onor, Massimo
Bramanti, Emilia
author_facet Campanella, Beatrice
Colombaioni, Laura
Nieri, Riccardo
Benedetti, Edoardo
Onor, Massimo
Bramanti, Emilia
author_sort Campanella, Beatrice
collection PubMed
description Metabolomic profiling of cell lines has shown many potential applications and advantages compared to animal models and human subjects, and an accurate cellular metabolite analysis is critical to understanding both the intracellular and extracellular environments in cell culture. This study provides a fast protocol to investigate in vitro metabolites of immortalized hippocampal neurons HN9.10e with minimal perturbation of the cell system using a targeted approach. HN9.10e neurons represent a reliable model of one of the most vulnerable regions of the central nervous system. Here, the assessment of their extracellular metabolic profile was performed by studying the cell culture medium before and after cell growth under standard conditions. The targeted analysis was performed by a direct, easy, high-throughput reversed-phase liquid chromatography with diode array detector (RP-HPLC-DAD) method and by headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GC-MS) for the study of volatile organic compounds (VOCs). The analysis of six different batches of cells has allowed to investigate the metabolic reproducibility of neuronal cells and to describe the metabolic “starting” conditions that are mandatory for a well-grounded interpretation of the results of any following cellular treatment. An accurate study of the metabolic profile of the HN9.10e cell line has never been performed before, and it could represent a quality parameter before any other targeting assay or further exploration.
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spelling pubmed-80856602021-05-01 Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions Campanella, Beatrice Colombaioni, Laura Nieri, Riccardo Benedetti, Edoardo Onor, Massimo Bramanti, Emilia Front Chem Chemistry Metabolomic profiling of cell lines has shown many potential applications and advantages compared to animal models and human subjects, and an accurate cellular metabolite analysis is critical to understanding both the intracellular and extracellular environments in cell culture. This study provides a fast protocol to investigate in vitro metabolites of immortalized hippocampal neurons HN9.10e with minimal perturbation of the cell system using a targeted approach. HN9.10e neurons represent a reliable model of one of the most vulnerable regions of the central nervous system. Here, the assessment of their extracellular metabolic profile was performed by studying the cell culture medium before and after cell growth under standard conditions. The targeted analysis was performed by a direct, easy, high-throughput reversed-phase liquid chromatography with diode array detector (RP-HPLC-DAD) method and by headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GC-MS) for the study of volatile organic compounds (VOCs). The analysis of six different batches of cells has allowed to investigate the metabolic reproducibility of neuronal cells and to describe the metabolic “starting” conditions that are mandatory for a well-grounded interpretation of the results of any following cellular treatment. An accurate study of the metabolic profile of the HN9.10e cell line has never been performed before, and it could represent a quality parameter before any other targeting assay or further exploration. Frontiers Media S.A. 2021-04-16 /pmc/articles/PMC8085660/ /pubmed/33937186 http://dx.doi.org/10.3389/fchem.2021.621548 Text en Copyright © 2021 Campanella, Colombaioni, Nieri, Benedetti, Onor and Bramanti. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Campanella, Beatrice
Colombaioni, Laura
Nieri, Riccardo
Benedetti, Edoardo
Onor, Massimo
Bramanti, Emilia
Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions
title Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions
title_full Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions
title_fullStr Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions
title_full_unstemmed Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions
title_short Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions
title_sort unraveling the extracellular metabolism of immortalized hippocampal neurons under normal growth conditions
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8085660/
https://www.ncbi.nlm.nih.gov/pubmed/33937186
http://dx.doi.org/10.3389/fchem.2021.621548
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