Summary of Study ST002285
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 PR001465. The data can be accessed directly via it's Project DOI: 10.21228/M8C12D 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 | ST002285 |
Study Title | Multiplatform mass spectrometry-based analysis of Leishmania donovani infected macrophages at different time points after infection |
Study Type | Mutiplatform mass spectrometry-based metabolomics, time course experiment |
Study Summary | The project aims to measure targeted and non-targeted metabolite data of intracellular extracts of uninfected and Leishmania-donovani infected macrophages at 0, 12, 36 and 72 hours post infection using a multiplatform mass spectrometry approach combining CE-TOF/MS (polar metabolites), LC-QTOF/MS (non-polar metabolites) and LC-QqQ/MS (polar metabolites) to characterize the dynamics of metabolic alterations ocurring in the human macrophage upon L. donovani infection. |
Institute | Universidad CEU San Pablo |
Department | Chemistry and Biochemistry |
Laboratory | Centre for Metabolomics and Bioanalysis (CEMBIO) |
Last Name | Fernández García |
First Name | Miguel |
Address | Centro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660 Boadilla del Monte. España |
mig.fernandez.ce@ceindo.ceu.es | |
Phone | +0034690090778 |
Submit Date | 2022-07-01 |
Raw Data Available | Yes |
Raw Data File Type(s) | mzML |
Analysis Type Detail | LC-MS |
Release Date | 2024-07-01 |
Release Version | 1 |
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Collection:
Collection ID: | CO002364 |
Collection Summary: | Cloned lines of Leishmania donovani (MHOM/IN/82/Patra1 and MHOM/ET/1967/Hu3) were maintained with weekly subpassages at 27 ºC in complete RPMI 1640 medium, supplemented with 10% heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin plus 100 mg/mL streptomycin and 20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer. Only parasites under ten passages were used in the experimental work. Peripheral blood mononuclear cells (PBMCs) were enriched from blood from healthy volunteers using a density gradient with Histopaque 1077. The mononuclear phase was recovered and washed with PBS. Next, cells were labeled with magnetic CD14 MicroBeads and CD14+ monocytes were positively separated using an MS column. Human CD14+ monocytes were differentiated in macrophages using recombinant macrophage colony stimulating factor 1 (M-CSF). Briefly, monocytes were plated at 1·106 cells/mL with 20 ng/mL of M-CSF. Growth factor was renewed at day 4 post-differentiation. The macrophages were used 7 days after differentiation. Macrophages were infected with L. donovani promastigotes at a 1:10 ratio. After 4 hours of incubation, non-phagocytosed parasites were removed, and cells were recovered. Macrophages were left in culture for 12, 36 and 72 h for metabolomic analysis. Uninfected macrophages were used as a control. To isolate uninfected and L. donovani-infected human monocyte-derived macrophages for further metabolomic analyses, cells were recovered, washed with phosphate buffer saline (PBS) and transferred into a previously weighted Eppendorf. After centrifugation the supernatant was discarded, and the pellet was immediately frozen in liquid nitrogen and stored at -80 ºC. |
Sample Type: | Cultured cells |