Summary of Study ST003879

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 PR002433. The data can be accessed directly via it's Project DOI: 10.21228/M84Z62 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST003879
Study Title	Establishing an insulin-sensitive human adipocyte culture model
Study Summary	To enable functional screens of insulin action in human fat cells, we generated adipocytes from adipose progenitor cells in vitro. Under standardized culture conditions, the adipocytes displayed limited insulin responsiveness. Our results prompted us to establish optimal insulin sensitizing conditions. One of the tested conditions resulted in fat cells with enhanced insulin responsiveness. To confirm that our results reflected expected effects in energy metabolism, we traced glucose conversion into central carbon metabolites following two hours of insulin stimulation. This revealed a robust increase in labeling of glycolytic and tricarboxylic acid (TCA) cycle intermediates in insulin-treated versus untreated cells.
Institute	Helmholtz Centre for Environmental Research
Last Name	Engelmann
First Name	Beatrice
Address	Permoserstr. 15
Email	beatrice.engelmann@ufz.de
Phone	00493412351099
Submit Date	2025-04-24
Raw Data Available	Yes
Raw Data File Type(s)	mzML, wiff
Analysis Type Detail	LC-MS
Release Date	2025-05-21
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002433
Project DOI:	doi: 10.21228/M84Z62
Project Title:	PLCXD1 as a Regulator of Insulin-Stimulated Lipid Metabolism in Human Adipocytes
Project Summary:	Insulin regulates adipocyte biology through transcriptional and post-transcriptional mechanisms, yet many insulin-responsive targets remain poorly characterized. Phospholipase C X domain-containing protein-1 (PLCXD1) emerged as a regulator of insulin-stimulated lipogenesis, without affecting lipolysis or adipogenesis. Our findings establish PLCXD1 as an insulin-responsive effector of lipid metabolism, with implications for adipocyte dysfunction in insulin resistance.
Institute:	Helmholtz Centre for Environmental Research
Last Name:	Engelmann
First Name:	Beatrice
Address:	Permoserstr. 15
Email:	beatrice.engelmann@ufz.de
Phone:	00493412351099

Subject:

Subject ID:	SU004014
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Male

Factors:

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

mb_sample_id	local_sample_id	Tretament
SA426635	21_Basal_sgCON	Basal
SA426636	13_Basal_sgCON	Basal
SA426637	14_Basal_sgCON	Basal
SA426638	17_Basal_sgCON	Basal
SA426639	18_Basal_sgCON	Basal
SA426640	22_Basal_sgCON	Basal
SA426641	2_Insulin_sgCON	Insulin
SA426642	9_Insulin_sgCON	Insulin
SA426643	5_Insulin_sgCON	Insulin
SA426644	10_Insulin_sgCON	Insulin
SA426645	1_Insulin_sgCON	Insulin
SA426646	6_Insulin_sgCON	Insulin

Showing results 1 to 12 of 12

Showing results 1 to 12 of 12

Collection:

Collection ID:	CO004007
Collection Summary:	Human preadipocytes were obtained from abdominal subcutaneous white adipose tissue (WAT) of a male patient.
Sample Type:	Subcutaneous white adipose tissue

Treatment:

Treatment ID:	TR004023
Treatment Summary:	Cells were cultured in 5% CO2 at 37°C. Cells were proliferated in DMEM with 10 mmol/l HEPES, 10% FBS, 50 μg/mL penicillin/streptomycin and 2.5 ng/mL Fibroblast growth factor-2 (FGF2). FGF was removed 24h prior to initiating differentiation. To initiate differentiation, media was switched to a 50/50 mixture of DMEM/F12 with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and insulin, triiodothyronine (T3), transferrin, dexamethasone, 3-isobutyl-1-methylxanthine (IBMX) and rosiglitazone for three days, after which dexamethasone and IBMX were removed. Rosiglitazone was removed on day 10 of differentiation. Various T3, transferrin and insulin supplementation conditions were compared for insulin sensitization, so that afterwards all experiments were performed with T3 and transferrin maintained in the media throughout differentiation, while insulin was reduced on day 10 of differentiation. On D13, cells were washed once with DMEM no glucose/glutamine supplemented with HEPES and PEST prior to incubation in DMEM with glucose and for 3h at 37°C. Cells were washed with DMEM no glucose/no glutamine and media replaced with tracing media containing labeled C13-glucose and unlabeled glutamine with or without insulin.

Treatment ID:

TR004023

Treatment Summary:

Cells were cultured in 5% CO2 at 37°C. Cells were proliferated in DMEM with 10 mmol/l HEPES, 10% FBS, 50 μg/mL penicillin/streptomycin and 2.5 ng/mL Fibroblast growth factor-2 (FGF2). FGF was removed 24h prior to initiating differentiation. To initiate differentiation, media was switched to a 50/50 mixture of DMEM/F12 with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and insulin, triiodothyronine (T3), transferrin, dexamethasone, 3-isobutyl-1-methylxanthine (IBMX) and rosiglitazone for three days, after which dexamethasone and IBMX were removed. Rosiglitazone was removed on day 10 of differentiation. Various T3, transferrin and insulin supplementation conditions were compared for insulin sensitization, so that afterwards all experiments were performed with T3 and transferrin maintained in the media throughout differentiation, while insulin was reduced on day 10 of differentiation. On D13, cells were washed once with DMEM no glucose/glutamine supplemented with HEPES and PEST prior to incubation in DMEM with glucose and for 3h at 37°C. Cells were washed with DMEM no glucose/no glutamine and media replaced with tracing media containing labeled C13-glucose and unlabeled glutamine with or without insulin.

Sample Preparation:

Sampleprep ID:	SP004020
Sampleprep Summary:	Extraction of intracellular and extracellular metabolites was performed by a 1:1:1 methanol:water:chloroform extraction protocol. For the extraction of intracellular metabolites, the culture medium was removed and the cells were rinsed twice with 1 ml of 0.9% ice-cold NaCl. The rinsing solution was removed, and the metabolism of the cells was stopped by adding 400 µL of MeOH (-20 °C) followed by 400 µL of ice-cold H2O containing 10 µM d6-glutarate. Cells were collected using a cell lifter and 400 µL of chloroform was added. After shaking at 1,400 rpm and 4 °C for 20 min, the extraction mixture was centrifuged at 18,000 g and 4 °C for 5 min. Subsequently, the polar upper phase was collected and evaporated to complete dryness.

Sampleprep ID:

SP004020

Sampleprep Summary:

Extraction of intracellular and extracellular metabolites was performed by a 1:1:1 methanol:water:chloroform extraction protocol. For the extraction of intracellular metabolites, the culture medium was removed and the cells were rinsed twice with 1 ml of 0.9% ice-cold NaCl. The rinsing solution was removed, and the metabolism of the cells was stopped by adding 400 µL of MeOH (-20 °C) followed by 400 µL of ice-cold H2O containing 10 µM d6-glutarate. Cells were collected using a cell lifter and 400 µL of chloroform was added. After shaking at 1,400 rpm and 4 °C for 20 min, the extraction mixture was centrifuged at 18,000 g and 4 °C for 5 min. Subsequently, the polar upper phase was collected and evaporated to complete dryness.

Chromatography:

Chromatography ID:	CH004834
Chromatography Summary:	Solvent A: 10mM tributylamine, 10mM acetic acid, 5% MeOH, 2% 2-propanol in water; Solvent B: 100% 2-propanol
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:	93% water/5% methanol/2% isopropanol; 10mM tributylamine; 10mM acetic acid
Solvent B:	100% isopropanol
Chromatography Type:	Reversed phase

Analysis:

Analysis ID:	AN006372
Analysis Type:	MS
Chromatography ID:	CH004834
Num Factors:	2
Num Metabolites:	76
Units:	Peak Area