Summary of Study ST002190

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 PR001396. The data can be accessed directly via it's Project DOI: 10.21228/M88H8K 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.

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Study ID	ST002190
Study Title	Mass Spectrometry Imaging of Lipids In A Gut Epithelial Cell Model
Study Type	Mass spectromery imaging of cells.
Study Summary	Scope: The Caco2/HT29-MTX co-culture system is widely used as a cell model of the intestinal epithelium. Although the gut epithelium plays an important role in the uptake of free fatty acids and the resynthesis of triglycerides the lipid distribution profile of the co-culture system is not well understood. Desorption electrospray ionization (DESI) is a mass spectrometry (MS) technique which has been widely used to study the main classes of lipid molecules on different tissue surfaces. This has been used to map lipid species and their distribution in Caco2 and HT29-MTX co-culture system. Methods and results: Caco2 and HT29-MTX cells were seeded on coverslips either singly or as cocultures in ratios of 75:25 and 50:50. Cells were cultured for 21 days before MS imaging using a DESI source in both the positive and negative ionization modes. The identity of selected lipids was confirmed in negative and positive ionisation modes using tandem MS. Although many lipids were common to both cell lines, there were distinctive patterns in the lipidomes. Thus, the lipidome of Caco2 cells was more heterogeneous and rich in cholesterol esters and triglycerides whilst HT29-MTX cells has a distinctive lipidome relating to phosphatidylethanolamines, phosphatidylinositols and odd chain lipids, including C17 fatty acids. Conclusion: DESI-MSI has shown that Caco2 and HT29-MTX cells have distinctive lipidomes which are still evident when the cells are cocultured. It has potential to both allow further validation of these widely used cell models and provide insights into how dietary components may modify lipid metabolism in future.
Institute	Manchester Institute of Biotechnology, University of Manchester
Last Name	Mattar
First Name	Hadeer
Address	Manchester Institute of Biotechnology, Princess Street, Manchester, UK, M1 7DN
Email	hmatar@bu.edu.sa
Phone	0161 306 6000
Submit Date	2022-05-11
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	MS(Dir. Inf.)
Release Date	2022-07-06
Release Version	1

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Treatment:

Treatment ID:	TR002288
Treatment Summary:	Caco2 and HT29-MTX cells were cultured separately in bulk in 25 cm2 flasks at 37 ºC, 5% CO2 for 2-3 days. DMEM complete media supplemented with 20% (v/v) foetal serum bovine, 1% (w/v) non-essential amino acids, 1% (w/v) L-glutamine and 1% (w/v) penicillin-streptomycin was used as culture medium. On reaching 80–90% confluency cells were trypsinised and then seeded on a coverslip at a density of 1x105 cells/ml either as separate cultures or in a co-culture system seeded at ratios of 75:25, 50:50 and 25:75 (Caco2: HT29-MTXcells). After seeding, each coverslip was placed in the well of a 6-well cell culture plate, and incubated at 37 ºC, 5% CO2. Media was changed every 48 h for 21 days until cells were confluent. Cells were prepared for DESI MSI analysis by rinsing coverslips in 150 mM ammonium acetate, pH 7.1 for 30 s before being allowed to dry in the air stream of a biological safety cabinet for 15 min. Coverslips were then thoroughly dried using a vacuum desiccator for 15 min before storage at -80 °C in petri dishes until required.

Treatment ID:

TR002288

Treatment Summary:

Caco2 and HT29-MTX cells were cultured separately in bulk in 25 cm2 flasks at 37 ºC, 5% CO2 for 2-3 days. DMEM complete media supplemented with 20% (v/v) foetal serum bovine, 1% (w/v) non-essential amino acids, 1% (w/v) L-glutamine and 1% (w/v) penicillin-streptomycin was used as culture medium. On reaching 80–90% confluency cells were trypsinised and then seeded on a coverslip at a density of 1x105 cells/ml either as separate cultures or in a co-culture system seeded at ratios of 75:25, 50:50 and 25:75 (Caco2: HT29-MTXcells). After seeding, each coverslip was placed in the well of a 6-well cell culture plate, and incubated at 37 ºC, 5% CO2. Media was changed every 48 h for 21 days until cells were confluent. Cells were prepared for DESI MSI analysis by rinsing coverslips in 150 mM ammonium acetate, pH 7.1 for 30 s before being allowed to dry in the air stream of a biological safety cabinet for 15 min. Coverslips were then thoroughly dried using a vacuum desiccator for 15 min before storage at -80 °C in petri dishes until required.