Return to study ST001035 main page
MB Sample ID: SA069330
| Local Sample ID: | MS6735-30 |
| Subject ID: | SU001074 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001074 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| MS6735-30 | SA069330 | FL011096 | 23h | Time |
| MS6735-30 | SA069330 | FL011096 | E Med | condition |
| MS6735-30 | SA069330 | FL011096 | Spent Sup | matrix |
| MS6735-30 | SA069330 | FL011096 | Unlabelled | Tracer |
Collection:
| Collection ID: | CO001068 |
| Collection Summary: | 4,000,000 CLOD cells collected by methanol scraping. 1ml sup also collected. There was no tracer element added to the sample. Also methanol extarction was done in ice. After the extraction samples were kept in dry ice and delivered to core. |
| Sample Type: | astrocytes |
Treatment:
| Treatment ID: | TR001088 |
| Treatment Summary: | Murine derived pure cortical astrocytes were plated at 4x10^6 density in 10cm petri dish and grown in astro growth medium (AGM) for 2-3 days until it reaches the 90% confluency. Day before the treatment with glutamine or glutamate AGM was changed to serum free “E” media. In case of isotomer analysis 40% of glucose was switched with U13C6 (glucose) and added to serum free “E” media. On the day of treatment first the spent media was collected and then washed with base media. After washing, astrocytes were exposed to different ratio of glutamate: glutamine for 1h under non-CO2 condition. 1h spent media was collected. After washing three times with phosphate buffer saline cell pellet was collected in ice cold 100% methanol by scrapping. All the samples were immediately transferred to -70C for future analysis. E media composition: DMEM (w/o phenol red)+ 1% PenStrep+ 10% FBS+ Glutamate+ Glucose Abbreviation: Gn-Glutamine Gt-Glutamate B- Base media G- Glucose P- Sodium pyruvate |
Sample Preparation:
| Sampleprep ID: | SP001081 |
| Sampleprep Summary: | 13C6 isotopomer analysis of Astrocyte We intend to test the experimental hypothesis that stimulation of primary murine astrocytes with patient-derived NMO IgG will drive a metabolic shift marked by alterations in cellular levels of glutamate and glutamine. To do so we will use isotopic tracing to measure glutamate and glutamine levels in cell extracts following stimulation with NMO IgG. The cell preparations, patient-derived antibody preps, isotopic tracer incubations, and cell stimulations will be performed in the PI’s lab. |
Combined analysis:
| Analysis ID | AN001696 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890B |
| Column | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Agilent 5977A |
| Ion Mode | POSITIVE |
| Units | Enrichment |
Chromatography:
| Chromatography ID: | CH001195 |
| Instrument Name: | Agilent 7890B |
| Column Name: | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS001571 |
| Analysis ID: | AN001696 |
| Instrument Name: | Agilent 5977A |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
