Summary of study ST001438

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000988. The data can be accessed directly via it's Project DOI: 10.21228/M80680 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001438
Study TitleSub-nanoliter metabolomics via mass spectrometry to characterize volume-limited samples - MSC
Study SummaryThe human metabolome provides a window into the mechanisms and biomarkers of various diseases. However, because of limited availability, many sample types are still difficult to study by metabolomic analyses. Here, we present a new mass spectrometry (MS)-based metabolomics strategy that only consumes sub-nanoliter sample volumes. The approach consists of combining a customized metabolomics workflow with a pulsed MS ion generation method, known as triboelectric nanogenerator inductive nanoelectrospray ionization (TENGi nanoESI) MS. A second example to illustrate TENGi MS capabilities involves rare cell metabolomics of cultured mesenchymal stromal cells (MSCs), a cell type that has shown potential for treating a variety of chronic diseases. Examination of metabolic changes of MSCs cultured under conditions that may impact in vitro therapeutic activity, such as aggregate culture, or preconditioning with interferon gamma (IFN- γ)13, is critical for identifying attributes of cell quality. Reducing cell numbers required to perform MSC metabolomic analysis is essential for improving the manufacturing of highly therapeutic MSCs without significantly impeding production.
Institute
Georgia Institute of Technology
Last NameFernandez
First NameFacundo
Address901 Atlantic Dr NE, Atlanta, GA, 30332, USA
Emailfernandez@gatech.edu
Phone404-385-4432
Submit Date2020-08-02
Raw Data AvailableYes
Raw Data File Type(s).raw
Analysis Type DetailESI
Release Date2020-09-14
Release Version1
Facundo Fernandez Facundo Fernandez
https://dx.doi.org/10.21228/M80680
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Treatment:

Treatment ID:TR001527
Treatment Summary:MSCs were then washed with PBS and trypsinized as described above, and the number of harvested cells counted using a hemocytometer. MSCs were then resuspended in 155 mM ammonium acetate (Fluka) at a concentration of 1.6x106 cells/mL and aliquoted into 50-µL samples (8x104 cells per aliquot). Cells were then quenched by adding 200-µL MeOH into each sample vial and stored at -80 C until metabolite extraction. Frozen cells were subject to three freeze-thaw cycles, with liquid nitrogen for freezing and ice-water sonication for thawing. Cell samples were then centrifuged at 14,800 rpm for 5 min to precipitate proteins. From the supernatant, 200 µL was transferred into a new vial for lyophilization. The pooled QC sample was formed by mixing 30 µL of each sample. All cell extracts and the QC sample were then lyophilized at -40 C and 100 mTorr for 24h in a VirTis Benchtop free-drier (LP Industries, Stone Ridge, NY, USA). Residues were reconstituted in a 5.9 ×10-5M 13C-phenylalanine methanolic solution to a final volume of 10 µL (for samples), and 18 µL (for QCs).
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