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MB Sample ID: SA184585
| Local Sample ID: | post-differentiation_later-1 |
| Subject ID: | SU002038 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 51.3±5 |
| Gender: | Female |
| Cell Biosource Or Supplier: | Promocell (GmbH, Heidelberg, Germany) and Zen-Bio (Zen-Bio, Inc., Research Triangle Park, NC, USA) |
| Cell Strain Details: | Human subcutaneous white adipocytes |
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Subject:
| Subject ID: | SU002038 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 51.3±5 |
| Gender: | Female |
| Cell Biosource Or Supplier: | Promocell (GmbH, Heidelberg, Germany) and Zen-Bio (Zen-Bio, Inc., Research Triangle Park, NC, USA) |
| Cell Strain Details: | Human subcutaneous white adipocytes |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| post-differentiation_later-1 | SA184585 | FL022693 | 1 | Cell Line |
| post-differentiation_later-1 | SA184585 | FL022693 | 5 | Stage |
Collection:
| Collection ID: | CO002031 |
| Collection Summary: | Three cell lines (Cell Line-1, Cell Line-2, and Cell Line-3) of Caucasian-derived subcutaneous preadipocytes were divided into five stages: subcutaneous preadipocytes (stage-1), after inducing differentiation into adipocytes (stage-2), after the initiation of fat accumulation, the early stage (stage-3), the middle stage (stage-4), and mature subcutaneous adipocytes (stage-5), respectively. |
| Sample Type: | Adipose tissue |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002050 |
| Treatment Summary: | Preadipocytes were grown in Promocell Preadipocyte Growth Medium until 100% confluent growth was observed (stage-1). Further, their differentiation into adipocytes was induced using Promocell Preadipocyte Differentiation Medium for 3 consecutive days. The post-differentiation adipocytes were designated as stage-2. After induction of differentiation, Promocell Preadipocyte Differentiation Medium was replaced with Promocell Adipocyte Nutrition Medium. The culture was continued for 7 days until the cytoplasm was filled with many lipid droplets, and the old medium was replaced with fresh Promocell Adipocyte Nutrition Medium after every 2 to 3 days during culturing. During this culture period, small lipid droplets began to appear in the cytoplasm (stage -3), followed by more lipid droplets (stage-4) and the cytoplasm filled with many lipid droplets (stage-5). |
Sample Preparation:
| Sampleprep ID: | SP002044 |
| Sampleprep Summary: | The cell suspension was transferred to a tube and centrifuged at 220g for 3 min, the supernatant was discarded and the cells were suspended in 100 μL of Milli-Q water. Each sample was normalized by measuring the amount of protein by bicinchoninic acid (BCA) protein assay using Pierce Micro BCA Protein Assay Kit (Pierce, Rockford, IL, USA) and the final volume of 600 μL was adjusted using water.10 μL of Splash Lipidomix internal standards (Avanti, Alabaster, AL, USA) was added and the Bligh and Dyer method was used for lipid extraction. |
| Processing Storage Conditions: | Room temperature |
| Extract Storage: | -20℃ |
Combined analysis:
| Analysis ID | AN003193 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Thermo Acclaim 120 C18 (150 x 2.1mm,3um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | normalized area values |
Chromatography:
| Chromatography ID: | CH002361 |
| Chromatography Summary: | We used a set of linear gradients starting at 20% of solvent B and increasing linearly to 100% over 50 min, maintained at 100% solvent B for 60 min, then decreased linearly to 20% B from 60 min to 60.1 min, and finished with 20% solvent B for the last 10 min. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Thermo Acclaim 120 C18 (150 x 2.1mm,3um) |
| Column Temperature: | 50 °C |
| Flow Rate: | 300 μL/min |
| Solvent A: | 50% water/25% methanol/25% acetonitrile; 0.1% formic acid; 5 mM ammonium formate |
| Solvent B: | 90% acetonitrile/ 10% water; 0.1% formic acid; 5 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002971 |
| Analysis ID: | AN003193 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Xcalibur software. LipidSearch for data processing. |
| Ion Mode: | UNSPECIFIED |
