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.

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Study IDST002285
Study TitleMultiplatform mass spectrometry-based analysis of Leishmania donovani infected macrophages at different time points after infection
Study TypeMutiplatform mass spectrometry-based metabolomics, time course experiment
Study SummaryThe 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
DepartmentChemistry and Biochemistry
LaboratoryCentre for Metabolomics and Bioanalysis (CEMBIO)
Last NameFernández García
First NameMiguel
AddressCentro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660 Boadilla del Monte. España
Emailmig.fernandez.ce@ceindo.ceu.es
Phone+0034690090778
Submit Date2022-07-01
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2024-07-01
Release Version1
Miguel Fernández García Miguel Fernández García
https://dx.doi.org/10.21228/M8C12D
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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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
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