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MB Sample ID: SA316365
Local Sample ID: | SGBS_ctrl_d0_1 |
Subject ID: | SU003027 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003027 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
SGBS_ctrl_d0_1 | SA316365 | FL037622 | d0 | Treatment |
SGBS_ctrl_d0_1 | SA316365 | FL037622 | 1 | Replicate |
SGBS_ctrl_d0_1 | SA316365 | FL037622 | Homo sapiens | Species |
Collection:
Collection ID: | CO003020 |
Collection Summary: | Human SGBS cells were provided by the laboratory of Prof. Dr. Wabitsch at the University Clinic Ulm. Cells were differentiated according to the standard protocol described previously (Wabitsch et al., 2001). 3T3-L1 mouse embryonic fibroblasts were purchased from the American Type Culture Collection (ATCC, CL-173, USA). |
Sample Type: | Adipose tissue |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR003036 |
Treatment Summary: | Both cell lines were maintained under 5% CO2 at 37°C and 95% humidity. Cell culture experiments for metabolomic (SGBS and 3T3L1) data analysis were performed in 6 replicates. Cells were cultivated until day0,day6 and day12 for the SGBS cell line, and until day0, day5 and day10 for the 3T3-L1 cells. |
Sample Preparation:
Sampleprep ID: | SP003033 |
Sampleprep Summary: | Intracellular metabolites were extracted using a 1:1:1 methanol:water:chloroform extraction protocol. Briefly, culture media was removed and cells were rinsed with icecold 0.9 % sodium chloride solution. The rinsing solution was removed and the cells metabolism was quenched by adding equal volumes of methanol, followed by icecold deionized water. Cells were scraped off the culture plate and combined chloroform. Samples were vortexed and kept on a shaker at 4°C for 20 min at 1400 rpm. Cell debris was removed by centrifugation. A fixed volume of the polar upper phase was transferred to new tubes and dried to completeness. |
Combined analysis:
Analysis ID | AN004784 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 Infinity II |
Column | Waters XSelect XP HSS T3 (150 x 2.1mm, 2.5um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex 6500+ |
Ion Mode | NEGATIVE |
Units | AUC |
Chromatography:
Chromatography ID: | CH003615 |
Instrument Name: | Agilent 1290 Infinity II |
Column Name: | Waters XSelect XP HSS T3 (150 x 2.1mm, 2.5um) |
Column Temperature: | 40 |
Flow Gradient: | 0-5 min 0% B, 5-9 min 0%- 2% B, 9-9.5 min 2-6% B, 9.5-11.5 min 6% B, 11.5-12 min 6-11% B, 12-13.5 min 11% B, 13.5-15.5 min 11-28% B, 15.5-16.5 min 28-53% B, 16.5-22.5 53% B, 22.5-23 min 53-0% B, 23-33 min 0% B |
Flow Rate: | 0-15.5 min 0.4 mL/min, 15.5-16.5 min 0.4-0.15 mL/min, 16.5-23 min 0.15 mL/min, 23-27 min 0.15-0.4 mL/min, 27-33 min 0.4 mL/min |
Solvent A: | 10mM TBA, 10mM acetic acid, 5% MeOH, 2% IPA in water |
Solvent B: | 100% IPA |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS004530 |
Analysis ID: | AN004784 |
Instrument Name: | ABI Sciex 6500+ |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | For identification and quantitation, a scheduled MRM method was used, with specific transitions for every metabolite. Data acquisition and peak integration were performed in Analyst® software (Version 1.7.1). |
Ion Mode: | NEGATIVE |