TY - JOUR
T1 - Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues
AU - Devilbiss, Andrew W.
AU - Zhao, Zhiyu
AU - Martin-Sandoval, Misty S.
AU - Ubellacker, Jessalyn M.
AU - Tasdogan, Alpaslan
AU - Agathocleous, Michalis
AU - Mathews, Thomas P.
AU - Morrison, Sean J.
N1 - Publisher Copyright:
© DeVilbiss et al.
PY - 2021/1
Y1 - 2021/1
N2 - 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.
AB - 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.
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U2 - 10.7554/eLife.61980
DO - 10.7554/eLife.61980
M3 - Article
C2 - 33470192
AN - SCOPUS:85100099532
SN - 2050-084X
VL - 10
SP - 1
EP - 23
JO - eLife
JF - eLife
M1 - e61980
ER -