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Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models
Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with some of the spatial and molecular complexity of tissue. For example, many cell lines form 3D structures given appropriate in vitro conditions. Colon...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4396945/ https://www.ncbi.nlm.nih.gov/pubmed/25549242 http://dx.doi.org/10.3791/52313 |
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author | Ahlf Wheatcraft, Dorothy R. Liu, Xin Hummon, Amanda B. |
author_facet | Ahlf Wheatcraft, Dorothy R. Liu, Xin Hummon, Amanda B. |
author_sort | Ahlf Wheatcraft, Dorothy R. |
collection | PubMed |
description | Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with some of the spatial and molecular complexity of tissue. For example, many cell lines form 3D structures given appropriate in vitro conditions. Colon cancer cell lines form 3D cell culture spheroids, in vitro mimics of avascular tumor nodules. While immunohistochemistry and other classical imaging methods are popular for monitoring the distribution of specific analytes, mass spectrometric imaging examines the distribution of classes of molecules in an unbiased fashion. While MALDI mass spectrometric imaging was originally developed to interrogate samples obtained from humans or animal models, this report describes the analysis of in vitro three dimensional cell cultures, including improvements in sample preparation strategies. Herein is described methods for growth, harvesting, sectioning, washing, and analysis of 3D cell cultures via matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) imaging. Using colon carcinoma 3D cell cultures as a model system, this protocol demonstrates the ability to monitor analytes in an unbiased fashion across the 3D cell culture system with MALDI-MSI. |
format | Online Article Text |
id | pubmed-4396945 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-43969452015-04-23 Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models Ahlf Wheatcraft, Dorothy R. Liu, Xin Hummon, Amanda B. J Vis Exp Bioengineering Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with some of the spatial and molecular complexity of tissue. For example, many cell lines form 3D structures given appropriate in vitro conditions. Colon cancer cell lines form 3D cell culture spheroids, in vitro mimics of avascular tumor nodules. While immunohistochemistry and other classical imaging methods are popular for monitoring the distribution of specific analytes, mass spectrometric imaging examines the distribution of classes of molecules in an unbiased fashion. While MALDI mass spectrometric imaging was originally developed to interrogate samples obtained from humans or animal models, this report describes the analysis of in vitro three dimensional cell cultures, including improvements in sample preparation strategies. Herein is described methods for growth, harvesting, sectioning, washing, and analysis of 3D cell cultures via matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) imaging. Using colon carcinoma 3D cell cultures as a model system, this protocol demonstrates the ability to monitor analytes in an unbiased fashion across the 3D cell culture system with MALDI-MSI. MyJove Corporation 2014-12-05 /pmc/articles/PMC4396945/ /pubmed/25549242 http://dx.doi.org/10.3791/52313 Text en Copyright © 2014, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Bioengineering Ahlf Wheatcraft, Dorothy R. Liu, Xin Hummon, Amanda B. Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models |
title | Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models |
title_full | Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models |
title_fullStr | Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models |
title_full_unstemmed | Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models |
title_short | Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models |
title_sort | sample preparation strategies for mass spectrometry imaging of 3d cell culture models |
topic | Bioengineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4396945/ https://www.ncbi.nlm.nih.gov/pubmed/25549242 http://dx.doi.org/10.3791/52313 |
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