Summary of Study ST001958

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001245. The data can be accessed directly via it's Project DOI: 10.21228/M8S985 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001958
Study TitleData on changes in lipid profiles during differentiation and maturation of human subcutaneous white adipocytes analyzed using chromatographic and bioinformatics tools
Study SummaryThree cell lines of Caucasian-derived subcutaneous preadipocytes were divided into five stages (stage-1 to stage-5) from subcutaneous preadipocytes to mature subcutaneous adipocytes filled with many lipid droplets. Lipids were extracted from cells in each stage and processed using untargeted liquid chromatography and Q-Exactive Orbitrap tandem mass spectrometry. The lipids were identified using LipidSearch 4.2.13.
Institute
Hamamatsu University School of Medicine
Last NameKitamoto
First NameTakuya
Address1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan
Emailt.ktmt@hama-med.ac.jp
Phone+81-53-435-2987
Submit Date2021-10-25
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2021-11-27
Release Version1
Takuya Kitamoto Takuya Kitamoto
https://dx.doi.org/10.21228/M8S985
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001245
Project DOI:doi: 10.21228/M8S985
Project Title:Data on changes in lipid profiles during differentiation and maturation of human subcutaneous white adipocytes analyzed using chromatographic and bioinformatics tools
Project Summary:Three cell lines of Caucasian-derived subcutaneous preadipocytes were divided into five stages (stage-1 to stage-5) from subcutaneous preadipocytes to mature subcutaneous adipocytes filled with many lipid droplets. Lipids were extracted from cells in each stage and processed using untargeted liquid chromatography and Q-Exactive Orbitrap tandem mass spectrometry. The lipids were identified using LipidSearch 4.2.13.
Institute:Hamamatsu University School of Medicine
Last Name:KITAMOTO
First Name:TAKUYA
Address:1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka, 431-3192, Japan
Email:t.ktmt@hama-med.ac.jp
Phone:+81-53-435-2987

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:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Cell Line Stage
SA184581preadipocytes-11 1
SA184582differentiation-11 2
SA184583post-differentiation_early-11 3
SA184584post-differentiation_middle-11 4
SA184585post-differentiation_later-11 5
SA184586preadipocytes-22 1
SA184587differentiation-22 2
SA184588post-differentiation_early-22 3
SA184589post-differentiation_middle-22 4
SA184590post-differentiation_later-22 5
SA184591preadipocytes-33 1
SA184592differentiation-33 2
SA184593post-differentiation_early-33 3
SA184594post-differentiation_middle-33 4
SA184595post-differentiation_later-33 5
Showing results 1 to 15 of 15

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
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