Summary of Study ST000403

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 PR000315. The data can be accessed directly via it's Project DOI: 10.21228/M8002N This work is supported by NIH grant, U2C- DK119886.

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Study IDST000403
Study TitleMetabolomics-based elucidation of active metabolic pathways in erythrocytes and HSC-derived reticulocytes
Study TypeCell type comparison
Study SummaryHuman stem cell derived reticulocytes were compared with mature erythrocytes by metabolomics analysis.
Institute
Monash Institute of Pharmaceutical Sciences
DepartmentDrug delivery, disposition and dynamics
LaboratoryCreek lab
Last NameCreek
First NameDarren
Address381 Royal Parade, Parkville, Melbourne, VIC3052, Australia
EmailDarren.Creek@monash.edu
PhoneN/A
Submit Date2016-05-11
Num Groups6
Total Subjects18
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2016-05-20
Release Version1
Darren Creek Darren Creek
https://dx.doi.org/10.21228/M8002N
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP000431
Sampleprep Summary:Metabolism was quenched by immersion of cultures in an ethanol/dry ice bath to 0-4 °C. Cells were pelleted by centrifugation (10,000 rpm for 1 min) and washed in cold PBS (1 mL). Metabolites were extracted from 1 x 108 RBCs and 0.5 x 108 reticulocytes by addition of 300 µL chloroform/methanol/water (1:3:1 v/v) containing internal standards (CHAPS, CAPS, PIPES and TRIS; 1 µM) and left for 30 mins at 4 °C with periodic mixing and sonication. The number of cells used for RBCs and reticulocytes was based on the mean cell volume of each cell type, and ensured that the total cell pellet volume was equivalent for each sample, allowing direct comparison of metabolite concentrations from these samples. After mixing, cellular debris was removed by centrifugation (16,000 rpm for 10 mins) and the supernatant was kept at -80 °C prior to analysis.
Processing Method:Lysis with mixing and sonication at 4°C
Processing Storage Conditions:on ice or 4°C
Extraction Method:chloroform/methanol/water (1:3:1 v/v)
Extract Storage:-80°C
Sample Spiking:internal standards (CHAPS, CAPS, PIPES and TRIS; all at 1 µM) mixed in extraction solvent
Cell Type:Human stem cell derived reticulocytes and mature erythrocytes
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