Summary of Study ST003492

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 PR002141. The data can be accessed directly via it's Project DOI: 10.21228/M8VZ4T This work is supported by NIH grant, U2C- DK119886.

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Study IDST003492
Study TitleMetabolomic profiling of the secretome in normal primary human pulmonary artery smooth muscle cells
Study SummaryGrowth medium of human pulmonary artery smooth muscle cells (PASMCs) was conditioned for 24 hours and analyzed by LC-MS, termed conditioned medium (CM). Comparing to fresh complete vascular smooth muscle cell growth medium (GM), LC-MS profiling screened 138 metabolites and identified that three branched chain alpha-ketoacids, namely KIC, KIV, and KMV are the most abundant metabolites in CM of human PASMCs.
Institute
Peking University
DepartmentDepartment of Toxicology
Last NameXiao
First NameWusheng
Address38 Xueyuan Rd, Haidian District, Beijing, Beijing, 100191, China
Emailwxiao@bjmu.edu.cn
Phone+1-010-82801628
Submit Date2023-08-07
Analysis Type DetailMALDI
Release Date2024-10-04
Release Version1
Wusheng Xiao Wusheng Xiao
https://dx.doi.org/10.21228/M8VZ4T
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002141
Project DOI:doi: 10.21228/M8VZ4T
Project Title:Secretome of normal primary human pulmonary artery smooth muscle cells
Project Summary:This project aims to delineate the possible mediators of aerobic activation of HIF1alpha in normal primary cells and its biological consequences related to pulmonary vascular pathobiology.
Institute:Peking University
Department:Department of Toxicology
Laboratory:WXIAO
Last Name:Xiao
First Name:Wusheng
Address:38 Xueyuan Rd, Haidian District, Beijing, Beijing, 100191, China
Email:wxiao@bjmu.edu.cn
Phone:+1-010-82801628

Subject:

Subject ID:SU003620
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Gender:Male and female
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id Treatment Sample source
SA384072CMfromSMC 04CM Vascular smooth muscle cells
SA384073CMfromSMC 05CM Vascular smooth muscle cells
SA384074CMfromSMC 06CM Vascular smooth muscle cells
SA384075SmGM 01GM Vascular smooth muscle cells
SA384076SmGM 02GM Vascular smooth muscle cells
SA384077SmGM 03GM Vascular smooth muscle cells
Showing results 1 to 6 of 6

Collection:

Collection ID:CO003613
Collection Summary:Human primary pulmonary artery smooth muscle cells were cultured in complete vascular smooth muscle cell growth medium in a T75 flask. When cells reach to roungly 80% confluence, growth medium were refreshed with 10 mL and cultured for 24 hours. Medium samples were then collected and used for LC-MS analysis after centrifugation and removal of cell debris.
Sample Type:Vascular smooth muscle cells
Storage Conditions:-80℃

Treatment:

Treatment ID:TR003629
Treatment Summary:Human primary pulmonary artery smooth muscle cells were cultured in complete vascular smooth muscle cell growth medium in a T75 flask. When cells reach to roungly 80% confluence, growth medium were refreshed with 10 mL and cultured for 24 hours. Medium samples were then collected (termed conditioned medium, CM) and used for LC-MS analysis after centrifugation and removal of cell debris. Regular complete growth medium (GM) of vascular smooth muscle cells was included for comparison.
Treatment:Conditioned medium (CM) vs regular growth medium (GM)
Treatment Compound:N/A
Treatment Route:N/A
Treatment Dose:N/A
Treatment Dosevolume:N/A
Treatment Doseduration:24 hours
Treatment Vehicle:N/A
Cell Growth Container:75 cm flasks
Cell Growth Config:1200 rpm
Cell Growth Rate:30-40 hours
Cell Media:Complete vascular smooth muscle cell growth medium supplemented with growth factors and 5% FBS
Cell Envir Cond:37 C and 5% CO2
Cell Pct Confluence:70-80%
Cell Media Lastchanged:Every 48 hours

Sample Preparation:

Sampleprep ID:SP003627
Sampleprep Summary:Metabolites in equal volume of GM and CM were extracted using 100% -80 degree prechilled LC-MS grade methanol. The final concentration of methanol was 80%. Proteins were removed by centrifugation and supernatants were collected and evaporate on an Integrated SpeedVac System to the starting volume (~200 uL). Twenty microliters of the resulting medium samples were loaded into a LC-MS sampling tube for analysis.
Processing Storage Conditions:Described in summary
Extract Storage:Described in summary

Combined analysis:

Analysis ID AN005732
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Vanquish
Column Merck ZIC-pHILIC(150 mm × 2.1 mm,3.5um)
MS Type MALDI
MS instrument type Triple quadrupole
MS instrument name Thermo Q Exactive Focus
Ion Mode UNSPECIFIED
Units ion intensity

Chromatography:

Chromatography ID:CH004351
Chromatography Summary:Vanquish ultra-high performance liquid chromatography system coupled to a Q Exactive mass spectrometer
Instrument Name:Thermo Vanquish
Column Name:Merck ZIC-pHILIC(150 mm × 2.1 mm,3.5um)
Column Temperature:4
Flow Gradient:gradient (%B) was 0 min, 80%; 20 min, 20%; 20.5 min, 80%; 28 min, 80%; and 42 min
Flow Rate:100
Solvent A:100% water; 20 mM ammonium carbonate; 0.1% ammonium hydroxide
Solvent B:100% acetonitrile
Chromatography Type:HILIC

MS:

MS ID:MS005455
Analysis ID:AN005732
Instrument Name:Thermo Q Exactive Focus
Instrument Type:Triple quadrupole
MS Type:MALDI
MS Comments:The mass spectrometer was operated in polarity-switching full scan mode from 70-1000 m/z. Resolution was set to 70,000, and the AGC target was 1×106 ions. Data were acquired and analyzed using TraceFinder 4.1 (Thermo) with peak identifications based on an in-house library of authentic metabolite standards previously analyzed utilizing this method. Missing values were imputed using random forest. Sample peak areas were normalized using probabilistic quotient normalization
Ion Mode:UNSPECIFIED
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