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.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

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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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Collection:

Collection ID:CO001507
Collection Summary:Bone marrow-derived MSCs (RoosterBio Inc., Lot #000139) were expanded for two passages in culture after being received. They were frozen in ~5x105 aliquots in Cryostor CS10 freeze media (BioLife). Frozen aliquots were revived and plated in tissue culture polystyrene flasks (Corning) for 3-4 days prior to seeding onto test surfaces. MSCs were cultured in low-glucose DMEM (Gibco) supplemented with 10% fetal bovine serum (FBS, Atlanta Biologicals, lot E16063), and 1% antibiotic/antimycotic solution (Gibco). Once confluent, MSCs were washed with sterile-filtered phosphate-buffered saline (PBS, Thermo Fisher) and detached from flasks using TrypLE express (Thermo Fisher). Dissociated cells were counted using a hemacytometer and replated at 13,000 cells/cm2 in T-75 tissue culture flasks. After overnight incubation, MSCs then were exposed to 48 hours of culture media (control conditions), or culture media supplemented with 50 ng/mL IFN- γ (Thermo Fisher). MSCs were then washed with PBS and trypsinized as described above, and the number of harvested cells counted using a hemocytometer.
Sample Type:Mesenchymal stromal cells
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