Summary of Study ST002743

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 PR001708. The data can be accessed directly via it's Project DOI: 10.21228/M8XM8H This work is supported by NIH grant, U2C- DK119886.

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Study IDST002743
Study TitleMetabolomics comparison of lung fibroblasts from Pteropus alecto and Homo sapiens
Study TypeMS Quantitative analysis
Study SummaryTargeted Metabolomics was performed to identify differences in central carbon metabolism between lung fibroblasts from black flying fox fruit bat Pteropus alecto (PaLung cells) and Humans (WI-38 cells). For both PaLung and WI-38 cells, metabolomics measurements were carried out for n = 3 biological replicates.
Institute
Duke-NUS Medical School
Last NameJagannathan
First NameNarendra Suhas
Address8 College Road, Singapore, Singapore, 169857, Singapore
Emailgmsnsja@nus.edu.sg
Phone(65) 66015073
Submit Date2023-06-25
Analysis Type DetailLC-MS
Release Date2024-06-27
Release Version1
Narendra Suhas Jagannathan Narendra Suhas Jagannathan
https://dx.doi.org/10.21228/M8XM8H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001708
Project DOI:doi: 10.21228/M8XM8H
Project Title:Metabolomics comparison of lung fibroblasts from Pteropus alecto and Homo sapiens
Project Type:MS quantitative analysis
Project Summary:Targeted Metabolomics was performed to identify differences in central carbon metabolism between lung fibroblasts from black flying fox fruit bat Pteropus alecto (PaLung cells) and Humans (WI-38 cells).
Institute:Duke-NUS Medical School
Last Name:Jagannathan
First Name:Narendra Suhas
Address:Centre for Computational Biology, 8 College Road, Singapore, Singapore, 169857, Singapore
Email:gmsnsja@nus.edu.sg
Phone:(65) 66015073
Project Comments:Metabolomics was performed by commissioning to Human Metabolome Technologies (HMT) service.
Contributors:Javier Yu Peng Koh, Yoko Itahana, Koji Itahana, Lisa Tucker-Kellogg

Subject:

Subject ID:SU002850
Subject Type:Cultured cells
Subject Species:Pteropus alecto ; Homo sapiens
Taxonomy ID:9402;9606
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id Genotype Species
SA289154WI38-1Wild-type H. sapiens
SA289155WI38-3Wild-type H. sapiens
SA289156WI38-2Wild-type H. sapiens
SA289157PaLung-3Wild-type P. alecto
SA289158PaLung-1Wild-type P. alecto
SA289159PaLung-2Wild-type P. alecto
Showing results 1 to 6 of 6

Collection:

Collection ID:CO002843
Collection Summary:WI-38 and PaLung cells were seeded in a 100 mm dish and harvested 2 days after seeding according to the protocol provided by Human Metabolome Technologies (HMT Japan).
Sample Type:Cultured fibroblasts

Treatment:

Treatment ID:TR002859
Treatment Summary:PaLung and WI-38 cells were washed with 5% (w/w) mannitol twice and fixed with methanol containing the internal standard solution. The internal standard solution was provided by HMT. Lysates were centrifuged at 2,300 x g for 5 min at 4°C, and supernatants were filtered using the provided filter units by centrifuging at 9,100 × g for 3 hours at 4 °C. Filtrates were evaporated by SpeedVac Concentrator SPD1010 (Thermo Fisher Scientific) and submitted to HMT for analysis. The total cell numbers at the time of harvest were about 1.4 x 107 cells for each cell line. Targeted quantitative analysis was performed by HMT using capillary electrophoresis mass spectrometry (CE-TOFMS and CE-QqQMS). Absolute abundances (normalized by cell numbers) were obtained for a total of 116 metabolites (54 and 62 metabolites in the cation and anion modes, respectively).

Sample Preparation:

Sampleprep ID:SP002856
Sampleprep Summary:PaLung and WI-38 cells were washed with 5% (w/w) mannitol twice and fixed with methanol containing the internal standard solution. The internal standard solution was provided by HMT. Lysates were centrifuged at 2,300 x g for 5 min at 4°C, and supernatants were filtered using the provided filter units by centrifuging at 9,100 × g for 3 hours at 4 °C. Filtrates were evaporated by SpeedVac Concentrator SPD1010 (Thermo Fisher Scientific) and submitted to HMT for analysis. The total cell numbers at the time of harvest were about 1.4 x 107 cells for each cell line. Targeted quantitative analysis was performed by HMT using capillary electrophoresis mass spectrometry (CE-TOFMS and CE-QqQMS). Absolute abundances (normalized by cell numbers) were obtained for a total of 116 metabolites (54 and 62 metabolites in the cation and anion modes, respectively).

Combined analysis:

Analysis ID AN004447 AN004448
Analysis type MS MS
Chromatography type CE CE
Chromatography system Agilent CE Agilent CE
Column Fused silica capillary, i.d. 50 μm × 80 cm Fused silica capillary, i.d. 50 μm × 80 cm
MS Type ESI ESI
MS instrument type TOF Triple quadrupole
MS instrument name Agilent CE-TOFMS Agilent 6460 QQQ
Ion Mode POSITIVE NEGATIVE
Units pmol/10^6 cells pmol/10^6 cells

Chromatography:

Chromatography ID:CH003341
Instrument Name:Agilent CE
Column Name:Fused silica capillary, i.d. 50 μm × 80 cm
Column Temperature:NA
Flow Gradient:NA
Flow Rate:NA
Solvent A:NA
Solvent B:NA
Chromatography Type:CE

MS:

MS ID:MS004194
Analysis ID:AN004447
Instrument Name:Agilent CE-TOFMS
Instrument Type:TOF
MS Type:ESI
MS Comments:Peaks detected in CE-TOFMS analysis were extracted using automatic integration software (MasterHands ver.2.18.0.1 developed at Keio University) (4) and those in CE-QqQMS analysis were extracted using automatic integration software (MassHunter Quantitative Analysis B.06.00 Agilent Technologies, Santa Clara, CA, USA) in order to obtain peak information, which includes m/z, migration time (MT), and peak area.
Ion Mode:POSITIVE
  
MS ID:MS004195
Analysis ID:AN004448
Instrument Name:Agilent 6460 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Peaks detected in CE-TOFMS analysis were extracted using automatic integration software (MasterHands ver.2.18.0.1 developed at Keio University) (4) and those in CE-QqQMS analysis were extracted using automatic integration software (MassHunter Quantitative Analysis B.06.00 Agilent Technologies, Santa Clara, CA, USA) in order to obtain peak information, which includes m/z, migration time (MT), and peak area.
Ion Mode:NEGATIVE
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