Summary of Study ST003166
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 PR001970. The data can be accessed directly via it's Project DOI: 10.21228/M8314P 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.
| Study ID | ST003166 |
| Study Title | Metabolomics of Murine WT or MCJ KO CD19-BBz CD8 CAR-T cells |
| Study Summary | MCJ/DnaJC15 is an endogenous negative regulator of Complex I and mitochondrial respiration. The goal of these experiments are to characterize the metabolic profile of murine WT or MCJ KO CD8 CD19-BBz CAR-T cells after 3 expansions (6 days) with IL-2 (60IU/ml). |
| Institute | University of Colorado School of Medicine |
| Laboratory | Laboratory of Angelo D'Alessandro in collaboration with Mercedes Rincon |
| Last Name | Cendali |
| First Name | Francesca |
| Address | 13199 East Montview Boulevard, Aurora, CO, 80045, USA |
| francesca.cendali@cuanschutz.edu | |
| Phone | 3037246131 |
| Submit Date | 2024-04-11 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-05-02 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO003278 |
| Collection Summary: | Mouse CD8 cells were isolated with negative selection as described above, activated with anti-CD3/anti-CD28 beads (Dynabeads™ Mouse T-Activator CD3/CD28 for T-Cell Expansion and Activation, Gibco, Cat#11453D) with addition of recombinant human IL-2 (rhIL-2) (60 IU/ml) and human recombinant IL-7 (10 ng/ml). On the second and third day of activation, the supernatant containing the CD19-BBz CAR/hEGFR retrovirus was spun on a plate coated with retronectin (Takara Bio Inc.) (2000xg for 2.5h at 32C). The activated cells were added to the viral-coated plate for transduction. The transduced CD8 CAR-T cells were removed from the beads on the fourth day and expanded with rhIL-2 (60 IU/ml) or rhIL-7 (10 ng/ml) and rhL-15 (100 ng/ml). After 2 days (1st expansion) cells were split 1/3 with fresh cytokine-containing medium. After 2 more days (2nd expansion), cells were split again with fresh cytokine-containing medium. Two days later (3rd expansion) cells were used for experiment or incubated in cytokine free-medium. For CAR+ cells enrichment, the CAR-T cells expanded with IL-2 for 3 expansions were incubated with an anti-hEGFR-PE (BioLegend, Cat#352904, RRID: AB_10896794) antibody, followed by incubation with anti-PE microbeads (Miltenyi, order#130-048-801), and purified using the Miltenyi LS columns as recommended by the manufacturer (Miltenyi). The enrichment resulted in a 98-100% CAR+ population, as verified by flow cytometry. |
| Sample Type: | T-cells |
