Summary of Study ST002469

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 PR001595. The data can be accessed directly via it's Project DOI: 10.21228/M8J13B This work is supported by NIH grant, U2C- DK119886.

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

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study ID	ST002469
Study Title	Mesenchymal stromal cell (MSC) Metabolite MS study
Study Type	Untargeted Metabolite Study
Study Summary	Metabolomics and lipidomics workflows were used to analyze Mesenchymal stromal cell (MSC) metabolites. Metabolite abundances were used to model MSC potency results in IDO and T-cell proliferation assays.
Institute	Georgia Institute of Technology
Department	Chemistry and Biochemistry
Laboratory	Fernandez Lab
Last Name	Van Grouw
First Name	Alexandria
Address	311 Ferst Dr. NW Atlanta, GA 30332
Email	agrouw3@gatech.edu
Phone	7072391412
Submit Date	2023-02-07
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2023-02-26
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP002565
Sampleprep Summary:	Approximately one million MSCs were analyzed for each sample. Frozen cell pellets were thawed and washed prior to undergoing a modified Bligh-Dyer extraction to yield two phases. Extraction solvent (2:2:1 chloroform:methanol:water) and glass beads (400-600 µm) were added to cell pellets for extraction and homogenization in a TissueLyser II to 30 Hz for 6 minutes. Samples were then sonicated and centrifuged. Following extraction, 300 µL aliquots from each layer were transferred to new microcentrifuge tubes and solvent was dried using vacuum centrifugation. Dried organic phase samples were re-constituted in isopropyl alcohol, while dried aqueous phase samples were re-constituted in 80% methanol. Re-constitution was followed by sonication, centrifugation, and transfer to liquid chromatography (LC) vials for ultrahigh performance liquid chromatography mass spectrometry (UHPLC-MS) analysis. Media samples without cells were also analyzed as blanks to remove any features corresponding to remaining media components on the cells. Ten µL of media was subject to the same Bligh-Dyer extraction as above and extracts were run according to the instrumental methods listed above. A quality control (QC) sample for hydrophilic interaction chromatography (HILIC) and reverse phase datasets was created by pooling 20 µL from each experimental sample. The pooled QC injections were used for drift correction of peak areas. Sample queue was randomized with a mix of samples, QCs, and blanks.

Sampleprep ID:

SP002565

Sampleprep Summary:

Approximately one million MSCs were analyzed for each sample. Frozen cell pellets were thawed and washed prior to undergoing a modified Bligh-Dyer extraction to yield two phases. Extraction solvent (2:2:1 chloroform:methanol:water) and glass beads (400-600 µm) were added to cell pellets for extraction and homogenization in a TissueLyser II to 30 Hz for 6 minutes. Samples were then sonicated and centrifuged. Following extraction, 300 µL aliquots from each layer were transferred to new microcentrifuge tubes and solvent was dried using vacuum centrifugation. Dried organic phase samples were re-constituted in isopropyl alcohol, while dried aqueous phase samples were re-constituted in 80% methanol. Re-constitution was followed by sonication, centrifugation, and transfer to liquid chromatography (LC) vials for ultrahigh performance liquid chromatography mass spectrometry (UHPLC-MS) analysis. Media samples without cells were also analyzed as blanks to remove any features corresponding to remaining media components on the cells. Ten µL of media was subject to the same Bligh-Dyer extraction as above and extracts were run according to the instrumental methods listed above. A quality control (QC) sample for hydrophilic interaction chromatography (HILIC) and reverse phase datasets was created by pooling 20 µL from each experimental sample. The pooled QC injections were used for drift correction of peak areas. Sample queue was randomized with a mix of samples, QCs, and blanks.