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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Treatment:

Treatment ID:	TR002571
Treatment Summary:	Bone marrow-derived MSCs (BMMSCs) were purchased from RoosterBio, Inc. (Frederick, MD), and iMSCs were purchased from Fujifilm Cellular Dynamics Inc (Madison, WI). Prior to this study’s expansion, MSCs were previously expanded to an initial population doubling level (PDL0). Cryopreserved vials from each donor were thawed, and 106 MSCs were seeded into an initial T-150 tissue culture flask in complete media containing Gibco™ Minimum Essential Media α with nucleosides (Thermo Fisher Scientific, Waltham, MA), 10% fetal bovine serum (FBS; HyClone Laboratories, Logan, UT), and 1% penicillin-streptomycin solution (10,000 U/mL; Sigma-Aldrich, St. Louis, MO) for a culture rescue period of 48 hr. The same lot of FBS was used throughout the study. MSCs were washed with endotoxin-free Dulbecco’s phosphate buffered saline (PBS) without calcium and magnesium (Millipore Sigma), harvested using 1X TrypLE™ Express Enzyme (Thermo Fisher Scientific), neutralized with complete media, and centrifuged 300g to create a cell pellet. MSCs were then resuspended in complete media and counted. Next, MSCs from each donor were seeded at 500 cells/cm2 into 10 T-75 flasks containing 10 mL complete media. Control flasks containing 10 mL complete media only were also prepared. All flasks were then transferred to a humidified incubator set to 37° C and 5% CO2. MSC conditioned medium (CM) sample collection of 300 µl was performed for each flask at approximately the same time each day (±1 hr) and total complete media exchange was performed every 3 days until MSCs achieved 70-80% confluence. All media samples were placed directly into –80° C storage until further analysis by NMR. MSCs were then harvested using the same procedure described above. Cell pellets were split for cryopreservation (and functional analysis, see below) or preparation for intracellular lipidomic/metabolic analysis. Cell pellets designated for cryopreservation were prepared into cryovials containing 106 MSCs in 1 mL CryoStor® CS 10 (Sigma-Aldrich) and stored at –80° C for 24 hr using controlled rate freezing containers. Vials were then transferred to the vapor phase in a liquid nitrogen cryogenic freezer until further analysis.

Treatment ID:

TR002571

Treatment Summary:

Bone marrow-derived MSCs (BMMSCs) were purchased from RoosterBio, Inc. (Frederick, MD), and iMSCs were purchased from Fujifilm Cellular Dynamics Inc (Madison, WI). Prior to this study’s expansion, MSCs were previously expanded to an initial population doubling level (PDL0). Cryopreserved vials from each donor were thawed, and 106 MSCs were seeded into an initial T-150 tissue culture flask in complete media containing Gibco™ Minimum Essential Media α with nucleosides (Thermo Fisher Scientific, Waltham, MA), 10% fetal bovine serum (FBS; HyClone Laboratories, Logan, UT), and 1% penicillin-streptomycin solution (10,000 U/mL; Sigma-Aldrich, St. Louis, MO) for a culture rescue period of 48 hr. The same lot of FBS was used throughout the study. MSCs were washed with endotoxin-free Dulbecco’s phosphate buffered saline (PBS) without calcium and magnesium (Millipore Sigma), harvested using 1X TrypLE™ Express Enzyme (Thermo Fisher Scientific), neutralized with complete media, and centrifuged 300g to create a cell pellet. MSCs were then resuspended in complete media and counted. Next, MSCs from each donor were seeded at 500 cells/cm2 into 10 T-75 flasks containing 10 mL complete media. Control flasks containing 10 mL complete media only were also prepared. All flasks were then transferred to a humidified incubator set to 37° C and 5% CO2. MSC conditioned medium (CM) sample collection of 300 µl was performed for each flask at approximately the same time each day (±1 hr) and total complete media exchange was performed every 3 days until MSCs achieved 70-80% confluence. All media samples were placed directly into –80° C storage until further analysis by NMR. MSCs were then harvested using the same procedure described above. Cell pellets were split for cryopreservation (and functional analysis, see below) or preparation for intracellular lipidomic/metabolic analysis. Cell pellets designated for cryopreservation were prepared into cryovials containing 106 MSCs in 1 mL CryoStor® CS 10 (Sigma-Aldrich) and stored at –80° C for 24 hr using controlled rate freezing containers. Vials were then transferred to the vapor phase in a liquid nitrogen cryogenic freezer until further analysis.