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MB Sample ID: SA238135
| Local Sample ID: | iC99-R1-007_2_Gln |
| Subject ID: | SU002478 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
| Cell Biosource Or Supplier: | ForIPS consortium |
| Cell Strain Details: | Midbrain neuronal precursor cells differentiated from patient derived induced pluripotent stem cells |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002478 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
| Cell Biosource Or Supplier: | ForIPS consortium |
| Cell Strain Details: | Midbrain neuronal precursor cells differentiated from patient derived induced pluripotent stem cells |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| iC99-R1-007_2_Gln | SA238135 | FL029589 | sPD | disease_state |
| iC99-R1-007_2_Gln | SA238135 | FL029589 | Glutamine | 13C_label |
Collection:
| Collection ID: | CO002471 |
| Collection Summary: | Neuronal precursor cells differentiated from patient derived induced pluripotent stem cells were seeded on Geltrex coated 6-well plates containing neuronal precursor maintenance medium at a density of 1,000,000 cells/well with six replicates per cell line. hNPCs were cultured at 37 °C, 7 % CO2, 21 % O2 for at least 72 h. |
| Sample Type: | Neurons |
| Storage Conditions: | -80? |
Treatment:
| Treatment ID: | TR002490 |
| Treatment Summary: | Growth medium was replaced by a labeling medium containing the respective stable-isotope tracer instead of its unlabeled variant. Cells were cultured with either 1.25 mM [U-13C]-glutamine or 21.25 mM [U-13C]-glucose for 24 h. |
Sample Preparation:
| Sampleprep ID: | SP002484 |
| Sampleprep Summary: | Subsequently, cell culture supernatant was stored for profiling the extracellular metabolome. Three replicates per cell line and blanks were washed with 0.9% NaCl and quenched with ice-cold methanol and ice-cold ddH2O (containing 1 µg/ml D6-glutaric acid as internal standard). Cells were scraped and extracts were added into tubes containing ice-cold chloroform. Following vortexing at 1,400 rpm for 20 min at 4°C and centrifugation at 17,000 g for 5 min at 4°C, 300 µl of the polar phase were transferred into GC glass vials with microinsert and dried under vacuum at 4°C. Dried extracts were derivatized using equal amounts of methoxylamine (20 mg/ml in pyridine) and MTBSTFA. |
Combined analysis:
| Analysis ID | AN003893 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890B gas chromatograph |
| Column | 30 m DB-35 ms and 5 m Duruguard capillary column |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Agilent 5977 |
| Ion Mode | POSITIVE |
| Units | Mass isotopomer distributions |
Chromatography:
| Chromatography ID: | CH002883 |
| Instrument Name: | Agilent 7890B gas chromatograph |
| Column Name: | 30 m DB-35 ms and 5 m Duruguard capillary column |
| Chromatography Type: | GC |
MS:
| MS ID: | MS003633 |
| Analysis ID: | AN003893 |
| Instrument Name: | Agilent 5977 |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | One microliter sample was injected into an SSL injector at 270 °C in a split or splitless mode. GC-MS analysis was performed using an Agilent 7890B gas chromatograph equipped with a 30 m DB-35 ms and 5 m Duruguard capillary column. Metabolites were detected in selected ion mode by an Agilent 5977 MSD system. The MetaboliteDetector software was used to analyze chromatograms, calculate mass isotopomer distributions (MIDs) and perform relative comparison of metabolite levels. |
| Ion Mode: | POSITIVE |
