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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000403 |
| Study Title | Metabolomics-based elucidation of active metabolic pathways in erythrocytes and HSC-derived reticulocytes |
| Study Type | Cell type comparison |
| Study Summary | Human stem cell derived reticulocytes were compared with mature erythrocytes by metabolomics analysis. |
| Institute | Monash Institute of Pharmaceutical Sciences |
| Department | Drug delivery, disposition and dynamics |
| Laboratory | Creek lab |
| Last Name | Creek |
| First Name | Darren |
| Address | 381 Royal Parade, Parkville, Melbourne, VIC3052, Australia |
| Darren.Creek@monash.edu | |
| Phone | N/A |
| Submit Date | 2016-05-11 |
| Num Groups | 6 |
| Total Subjects | 18 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2016-05-20 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO000418 |
| Collection Summary: | Peripheral blood mononucleated cells were obtained from blood by Percoll density purification and CD34+ hemopoietic progenitor cells were isolated by magnetic bead separation according to the manufacturer's instructions (Miltenyi Biotec). CD34+ cells were cultured in a three-stage protocol based on the methods of 2d. Initially cells were cultured at 37 °C in a humid atmosphere of 5% CO2 at a density of 1 x 104 cells/mL and then maintained in the range of 2-10 × 105 cells/mL in IMDM (LifeTech) containing 5% (v/v) AB Serum (Interstate Companies Laboratories), 10 μg/mL Insulin (Sigma), 3 U/mL heparin (Pfizer), 200 μg/mL Transferrin (Prospec), 3 U/mL EPO (Eprex). During stage one (days 0-8) this was supplemented with 10 ng/mL SCF (GenScript) and 1 ng/mL IL-3 (R&D systems); during stage two (days 8-11) with 10 ng/mL SCF and additional 800 μg/mL transferrin and stage 3 (days 11-18) with 3 U/mL EPO and additional 800 μg/mL transferrin. Cultured reticulocytes (cRetics) were filtered at day 18 using a PALL WBF leukocyte filter. Isogenic control red blood cells (RBCs) were retained from donor blood, washed in IMDM and stored in saline-adenine-glucose-mannitol solution (SAG-M) at 4°C prior to use. Before analysis, cells were washed and cultured overnight in stage 3-supplemented IMDM (as outlined above). |
| Sample Type: | Cells |
| Collection Method: | See summary |
