Summary of Study ST004430

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 PR002673. The data can be accessed directly via it's Project DOI: 10.21228/M84P1D This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST004430
Study Title	Time-resolved lipidomics analysis of TERT-hWA adipocytes upon differentiation
Study Summary	TERT-hWA cells (5,000 per well) were seeded into ultra-low attachment plates in medium containing 2% FBS and allowed to form spheroids for 5 days. On day 0, pre-formed spheroids were treated with an adipogenic differentiation cocktail and cultured for an additional 2 weeks to generate fully differentiated adipocyte spheroids. At defined days of the differentiation process (days 0, 6, 12, and 19), eight spheroids per sample were collected, and lipids were extracted. Lipids were analyzed by LC-MS/MS in the resulted ACN and HEX fractions.
Institute	University of Szeged
Last Name	David
First Name	Kovacs
Address	Dóm tér 9, Szeged, Csongrád-Csanád, 6723, Hungary
Email	kovacs.david@med.u-szeged.hu
Phone	+3662342665
Submit Date	2025-12-03
Raw Data Available	Yes
Raw Data File Type(s)	mzML, raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2026-01-02
Release Version	1

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Project:

Project ID:	PR002673
Project DOI:	doi: 10.21228/M84P1D
Project Title:	Chain length-dependent mobilization and oxidation of fatty acids in adipocytes
Project Summary:	Fatty acids (FAs) are essential metabolites in energy homeostasis. Adipocytes store FAs as triacylglycerol (TG) in their lipid droplets and mobilize them upon demand in order to provide energy for peripheral tissues. While the mobilization of long-chain fatty acids (LCFAs) during lipolysis is largely documented, there is little information on the metabolism of short- and medium-chain fatty acids (SMCFA) in adipocytes. We demonstrate that adipocytes store SMCFAs in their lipid droplets and that following lipolysis initiation, TGs containing SMCFAs undergo rapid hydrolysis. We found that this process is facilitated by the preferential accumulation of SMCFA-containing TGs at the surface of lipid droplets, thereby enhancing their accessibility to lipases. Unlike LCFAs, SMCFAs are not released from the adipocytes following lipolysis but undergo oxidation within the cell. Our findings suggest that SMCFAs are preferentially mobilized and oxidized to provide energy for the adipocyte, highlighting a distinct metabolic fate compared to LCFAs.
Institute:	University of Szeged
Department:	Institute of Biochemistry
Last Name:	Kovacs
First Name:	David
Address:	Dóm tér 9, Szeged, Csongrád-Csanád, 6723, Hungary
Email:	kovacs.david@med.u-szeged.hu
Phone:	+3662342665

Subject:

Subject ID:	SU004591
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

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

mb_sample_id	local_sample_id	Condition	Injection order	Sample source
SA523710	37	Blank	1	PBS
SA523711	76	Blank	1	PBS
SA523712	77	Blank	2	PBS
SA523713	38	Blank	2	PBS
SA523714	78	Blank	3	PBS
SA523715	39	Blank	3	PBS
SA523716	46	Day_0	1	Adipocyte_spheroids
SA523717	43	Day_0	1	Adipocyte_spheroids
SA523718	1	Day_0	1	Adipocyte_spheroids
SA523719	40	Day_0	1	Adipocyte_spheroids
SA523720	4	Day_0	1	Adipocyte_spheroids
SA523721	7	Day_0	1	Adipocyte_spheroids
SA523722	5	Day_0	2	Adipocyte_spheroids
SA523723	47	Day_0	2	Adipocyte_spheroids
SA523724	44	Day_0	2	Adipocyte_spheroids
SA523725	41	Day_0	2	Adipocyte_spheroids
SA523726	2	Day_0	2	Adipocyte_spheroids
SA523727	8	Day_0	2	Adipocyte_spheroids
SA523728	9	Day_0	3	Adipocyte_spheroids
SA523729	3	Day_0	3	Adipocyte_spheroids
SA523730	48	Day_0	3	Adipocyte_spheroids
SA523731	6	Day_0	3	Adipocyte_spheroids
SA523732	42	Day_0	3	Adipocyte_spheroids
SA523733	45	Day_0	3	Adipocyte_spheroids
SA523734	10	Day_12	1	Adipocyte_spheroids
SA523735	13	Day_12	1	Adipocyte_spheroids
SA523736	55	Day_12	1	Adipocyte_spheroids
SA523737	52	Day_12	1	Adipocyte_spheroids
SA523738	49	Day_12	1	Adipocyte_spheroids
SA523739	16	Day_12	1	Adipocyte_spheroids
SA523740	11	Day_12	2	Adipocyte_spheroids
SA523741	14	Day_12	2	Adipocyte_spheroids
SA523742	56	Day_12	2	Adipocyte_spheroids
SA523743	50	Day_12	2	Adipocyte_spheroids
SA523744	17	Day_12	2	Adipocyte_spheroids
SA523745	53	Day_12	2	Adipocyte_spheroids
SA523746	54	Day_12	3	Adipocyte_spheroids
SA523747	15	Day_12	3	Adipocyte_spheroids
SA523748	12	Day_12	3	Adipocyte_spheroids
SA523749	18	Day_12	3	Adipocyte_spheroids
SA523750	57	Day_12	3	Adipocyte_spheroids
SA523751	51	Day_12	3	Adipocyte_spheroids
SA523752	25	Day_19	1	Adipocyte_spheroids
SA523753	22	Day_19	1	Adipocyte_spheroids
SA523754	64	Day_19	1	Adipocyte_spheroids
SA523755	19	Day_19	1	Adipocyte_spheroids
SA523756	61	Day_19	1	Adipocyte_spheroids
SA523757	58	Day_19	1	Adipocyte_spheroids
SA523758	65	Day_19	2	Adipocyte_spheroids
SA523759	62	Day_19	2	Adipocyte_spheroids
SA523760	59	Day_19	2	Adipocyte_spheroids
SA523761	26	Day_19	2	Adipocyte_spheroids
SA523762	23	Day_19	2	Adipocyte_spheroids
SA523763	20	Day_19	2	Adipocyte_spheroids
SA523764	60	Day_19	3	Adipocyte_spheroids
SA523765	21	Day_19	3	Adipocyte_spheroids
SA523766	63	Day_19	3	Adipocyte_spheroids
SA523767	27	Day_19	3	Adipocyte_spheroids
SA523768	24	Day_19	3	Adipocyte_spheroids
SA523769	66	Day_19	3	Adipocyte_spheroids
SA523770	70	Day_6	1	Adipocyte_spheroids
SA523771	34	Day_6	1	Adipocyte_spheroids
SA523772	28	Day_6	1	Adipocyte_spheroids
SA523773	31	Day_6	1	Adipocyte_spheroids
SA523774	67	Day_6	1	Adipocyte_spheroids
SA523775	73	Day_6	1	Adipocyte_spheroids
SA523776	29	Day_6	2	Adipocyte_spheroids
SA523777	74	Day_6	2	Adipocyte_spheroids
SA523778	71	Day_6	2	Adipocyte_spheroids
SA523779	35	Day_6	2	Adipocyte_spheroids
SA523780	68	Day_6	2	Adipocyte_spheroids
SA523781	32	Day_6	2	Adipocyte_spheroids
SA523782	72	Day_6	3	Adipocyte_spheroids
SA523783	69	Day_6	3	Adipocyte_spheroids
SA523784	75	Day_6	3	Adipocyte_spheroids
SA523785	30	Day_6	3	Adipocyte_spheroids
SA523786	33	Day_6	3	Adipocyte_spheroids
SA523787	36	Day_6	3	Adipocyte_spheroids

Showing results 1 to 78 of 78

Showing results 1 to 78 of 78

Collection:

Collection ID:	CO004584
Collection Summary:	On the indicated days of differentiation, spheroids were pooled in a minimal volume of PBS in glass tubes (8 spheroid/replicate) then ACN and Hex phases were extracted from TERT-hWA as described in the Sampleprep section. The phases were analyzed by LC-MS/MS and each sample was injected at least times.
Sample Type:	Adipocyte spheroids

Treatment:

Treatment ID:	TR004600
Treatment Summary:	Pooled spheroids (8/sample) at different days of differentiation (Day 0, 6, 12 and 19) were collected and extracted.

Sample Preparation:

Sampleprep ID:	SP004597
Sampleprep Summary:	To extract lipids from spheroids, a 3-phase liquid extraction was performed to separate neutral and polar lipids. Spheroids were collected in a minimum phosphate-buffered saline (PBS) volume (aqueous). Spheroids were transferred into glass tubes, containing already the internal standard mix (Splash) and TG 6:0/6:0/6:0. Then, to reduce sample loss, 0.75 mL of ACN were added to microtubes, vortexed, and transferred into the corresponding glass tubes. The remaining solvents were added to each sample: 0.75 mL Hex, 0.25 mL EtAc. The aqueous phase (sample) was completed with ultrapure water when needed, resulting in Hex:EtAc:ACN:Aqueous (3:1:3:2, V:V:V:V). Spheroid samples were first vortexed for 30 min at 4oC with glass beads, and only then centrifuged. The upper phase was collected into a new tube with a Hamilton glass syringe, which was washed three times in solvent between samples. Hex was added (half the volume of the first extraction) to the 2 remaining phases for re-extraction. Samples were again vortexed and centrifuged, and upper and middle phases were collected separately. To reduce phospholipid loss, a re-extraction of middle phase was done with ACN:EtAc (3:1, V:V) (half the volume of first extraction). Extraction (solvents and water) and experiment blanks (PBS) were included. All extraction solvents had 50 μg/mL BHT. Extracted samples were kept dried at -20oC under Ar.

Sampleprep ID:

SP004597

Sampleprep Summary:

To extract lipids from spheroids, a 3-phase liquid extraction was performed to separate neutral and polar lipids. Spheroids were collected in a minimum phosphate-buffered saline (PBS) volume (aqueous). Spheroids were transferred into glass tubes, containing already the internal standard mix (Splash) and TG 6:0/6:0/6:0. Then, to reduce sample loss, 0.75 mL of ACN were added to microtubes, vortexed, and transferred into the corresponding glass tubes. The remaining solvents were added to each sample: 0.75 mL Hex, 0.25 mL EtAc. The aqueous phase (sample) was completed with ultrapure water when needed, resulting in Hex:EtAc:ACN:Aqueous (3:1:3:2, V:V:V:V). Spheroid samples were first vortexed for 30 min at 4oC with glass beads, and only then centrifuged. The upper phase was collected into a new tube with a Hamilton glass syringe, which was washed three times in solvent between samples. Hex was added (half the volume of the first extraction) to the 2 remaining phases for re-extraction. Samples were again vortexed and centrifuged, and upper and middle phases were collected separately. To reduce phospholipid loss, a re-extraction of middle phase was done with ACN:EtAc (3:1, V:V) (half the volume of first extraction). Extraction (solvents and water) and experiment blanks (PBS) were included. All extraction solvents had 50 μg/mL BHT. Extracted samples were kept dried at -20oC under Ar.

Combined analysis:

Analysis ID	AN007411
Chromatography ID	CH005615
MS ID	MS007103
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Thermo Dionex Ultimate 3000
Column	Thermo Accucore C18 (150 x 2.1 mm, 2.6 µm)
MS Type	ESI
MS instrument type	Orbitrap
MS instrument name	Thermo Q Exactive Orbitrap
Ion Mode	POSITIVE
Units	raw intensity

Chromatography:

Chromatography ID:	CH005615
Instrument Name:	Thermo Dionex Ultimate 3000
Column Name:	Thermo Accucore C18 (150 x 2.1 mm, 2.6 µm)
Column Temperature:	35°C
Flow Gradient:	0.0 min, 35% B; 4.0 min, 60% B; 8.0 min, 70% B; 16.0 min, 85% B; 25.0 min, 97% B
Flow Rate:	400 µL/min
Solvent A:	50% Acetonitrile/50% Water; 10 mM ammonium formate; 0.1% formic acid
Solvent B:	88% Isopropanol/10% Acetonitrile/2% Water; 2 mM ammonium formate; 0.02 % formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS007103
Analysis ID:	AN007411
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	Data acquisition was conducted in positive ionization mode using Xcalibur software (v4.1.31.9). Full scan MS spectra were acquired in data-dependent MS2 (dd-MS2) mode at a resolution of 70 000 (at m/z 200) over a mass range of m/z 250-1200. The automatic gain control (AGC) target was set to 1 × 10⁶ with a maximum injection time of 100 ms. The 15 most intense precursor ions per sac cycle were selected for fragmentation. Precursor isolation was performed with a 1.0 m/z isolation window, and fragmentation was achieved using higher-energy collisional dissociation (HCD) at normalized collision energy (NCE) of 25 and 30 eV. MS/MS spectra were acquired in the ion trap at a resolution of 35,000 (at m/z 200) with an AGC target of 1 × 10⁵ and a maximum injection time of 80 ms.
Ion Mode:	POSITIVE