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Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues
Little is known about the metabolic regulation of rare cell populations because most metabolites are hard to detect in small numbers of cells. We previously described a method for metabolomic profiling of flow cytometrically isolated hematopoietic stem cells (HSCs) that detects 60 metabolites in 10,...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7847306/ https://www.ncbi.nlm.nih.gov/pubmed/33470192 http://dx.doi.org/10.7554/eLife.61980 |
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author | DeVilbiss, Andrew W Zhao, Zhiyu Martin-Sandoval, Misty S Ubellacker, Jessalyn M Tasdogan, Alpaslan Agathocleous, Michalis Mathews, Thomas P Morrison, Sean J |
author_facet | DeVilbiss, Andrew W Zhao, Zhiyu Martin-Sandoval, Misty S Ubellacker, Jessalyn M Tasdogan, Alpaslan Agathocleous, Michalis Mathews, Thomas P Morrison, Sean J |
author_sort | DeVilbiss, Andrew W |
collection | PubMed |
description | Little is known about the metabolic regulation of rare cell populations because most metabolites are hard to detect in small numbers of cells. We previously described a method for metabolomic profiling of flow cytometrically isolated hematopoietic stem cells (HSCs) that detects 60 metabolites in 10,000 cells (Agathocleous et al., 2017). Here we describe a new method involving hydrophilic liquid interaction chromatography and high-sensitivity orbitrap mass spectrometry that detected 160 metabolites in 10,000 HSCs, including many more glycolytic and lipid intermediates. We improved chromatographic separation, increased mass resolution, minimized ion suppression, and eliminated sample drying. Most metabolite levels did not significantly change during cell isolation. Mouse HSCs exhibited increased glycerophospholipids relative to bone marrow cells and methotrexate treatment altered purine biosynthesis. Circulating human melanoma cells were depleted for purine intermediates relative to subcutaneous tumors, suggesting decreased purine synthesis during metastasis. These methods facilitate the routine metabolomic analysis of rare cells from tissues. |
format | Online Article Text |
id | pubmed-7847306 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-78473062021-02-01 Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues DeVilbiss, Andrew W Zhao, Zhiyu Martin-Sandoval, Misty S Ubellacker, Jessalyn M Tasdogan, Alpaslan Agathocleous, Michalis Mathews, Thomas P Morrison, Sean J eLife Stem Cells and Regenerative Medicine Little is known about the metabolic regulation of rare cell populations because most metabolites are hard to detect in small numbers of cells. We previously described a method for metabolomic profiling of flow cytometrically isolated hematopoietic stem cells (HSCs) that detects 60 metabolites in 10,000 cells (Agathocleous et al., 2017). Here we describe a new method involving hydrophilic liquid interaction chromatography and high-sensitivity orbitrap mass spectrometry that detected 160 metabolites in 10,000 HSCs, including many more glycolytic and lipid intermediates. We improved chromatographic separation, increased mass resolution, minimized ion suppression, and eliminated sample drying. Most metabolite levels did not significantly change during cell isolation. Mouse HSCs exhibited increased glycerophospholipids relative to bone marrow cells and methotrexate treatment altered purine biosynthesis. Circulating human melanoma cells were depleted for purine intermediates relative to subcutaneous tumors, suggesting decreased purine synthesis during metastasis. These methods facilitate the routine metabolomic analysis of rare cells from tissues. eLife Sciences Publications, Ltd 2021-01-20 /pmc/articles/PMC7847306/ /pubmed/33470192 http://dx.doi.org/10.7554/eLife.61980 Text en © 2021, DeVilbiss et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Stem Cells and Regenerative Medicine DeVilbiss, Andrew W Zhao, Zhiyu Martin-Sandoval, Misty S Ubellacker, Jessalyn M Tasdogan, Alpaslan Agathocleous, Michalis Mathews, Thomas P Morrison, Sean J Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
title | Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
title_full | Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
title_fullStr | Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
title_full_unstemmed | Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
title_short | Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
title_sort | metabolomic profiling of rare cell populations isolated by flow cytometry from tissues |
topic | Stem Cells and Regenerative Medicine |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7847306/ https://www.ncbi.nlm.nih.gov/pubmed/33470192 http://dx.doi.org/10.7554/eLife.61980 |
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