Return to study ST001023 main page
MB Sample ID: SA064316
| Local Sample ID: | Sample # 1 |
| Subject ID: | SU001062 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001062 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Sample # 1 | SA064316 | FL010958 | Cells | Group |
| Sample # 1 | SA064316 | FL010958 | DIPG XIII | cell line |
Collection:
| Collection ID: | CO001056 |
| Collection Summary: | DIPG XIII and DIPG XVII cell lines are collected in this experiment. Susupension cells are harvested using centrifugation at 1200 rpm for 5 min. 1 mL of the supernatant media werr collected in an eppendorf tube and snapped frozen. The cell pellets were broken up into single cell suspension and counted. 1 million cells were taken from the stock and washed 1 x with PBS using table top centrifuge with 10 sec quick spin. The resulting cell pellet was snap frozen. Both the frozen media and cell pellet are stored in -80 C prior transfer. |
| Sample Type: | Glioma cells |
Treatment:
| Treatment ID: | TR001076 |
| Treatment Summary: | Metabolic profiling will be conducted similarly to published methods and standard methods from our metabolic core. Glucose, glutamine and lactate levels in culture medium will be measured using gas chromatography/mass spectroscopy (GC/MS). Briefly, cells will be seeded in 6 well plates in triplicate using our standard neurosphere media (MH+++) and cultured for 24 to 48h before experiments. Changes in metabolite concentrations relative to fresh media will be normalized to protein content of each well. Cellular metabolite levels will be measured using standard protocols using deuterated 2-hydroxyglutarate (d5-5HG) as an internal standard and analyzed using GC/MS. To determine the fraction of metabolites from Glu and Gln, 13C versions of each metabolite [U-13C]glucose or [U-13C]glutamine (Cambridge Isotope Lab) will be used. All of these experiements will be performed by the Mayo Clinic Metabolomics Research Core. |
Sample Preparation:
| Sampleprep ID: | SP001069 |
| Sampleprep Summary: | TCA Concentrations in glioma cell lines |
Combined analysis:
| Analysis ID | AN001680 |
|---|---|
| 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 | nmol/vial |
Chromatography:
| Chromatography ID: | CH001181 |
| Instrument Name: | Agilent 7890B |
| Column Name: | Agilent HP5-MS (30m × 0.25mm, 0.25 um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS001555 |
| Analysis ID: | AN001680 |
| Instrument Name: | Agilent 5977A |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
