Summary of Study ST001622
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 PR001041. The data can be accessed directly via it's Project DOI: 10.21228/M84M6M This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001622 |
| Study Title | Isotope Tracing Analysis in Intestinal Organoids with fructose |
| Study Summary | We isolated and cultured Intestinal Organoids from Mouse. To trace the metabolism of fructose, intestinal organoids were cultured in unlabeled fructose-containing DMEM/F12 medium (10 mM D-fructose, 0 mM D-glucose, 2.5 mM L-glutamine) for 12 hr, and then switched into 13C labeled fructose-containing medium (10 mM U-[13C] D-fructose, Sigma-aldrich) for 6 hr. |
| Institute | China Pharmaceutical University |
| Department | School of Medicine |
| Laboratory | Metabonomics |
| Last Name | Hou |
| First Name | Yuanlong |
| Address | tongjiaxiang, nanjing, jiangsu, 210000, China |
| jian2103@163.com | |
| Phone | 18851105337 |
| Submit Date | 2020-12-09 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Waters) |
| Analysis Type Detail | LC-MS |
| Release Date | 2020-12-16 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001041 |
| Project DOI: | doi: 10.21228/M84M6M |
| Project Title: | Dietary composition analysis studies |
| Project Type: | Untargeted metabolomics analysis with two diets |
| Project Summary: | Dietary patterns and psychosocial factors, ubiquitous part of modern lifestyle, critically shape the gut microbiota and human health. However, it remains obscure how dietary and psychosocial inputs coordinately modulate the gut microbiota and host impact. Here, we show that dietary raffinose metabolism to fructose couples stress-induced gut microbial remodeling to intestinal stem cells (ISC) renewal and epithelial homeostasis. Chow diet (CD) and purified diet (PD) confer distinct vulnerability to gut epithelial injury, microbial alternation and ISC dysfunction in chronically restrained mice. CD preferably enriches Lactobacillus reuteri, and its colonization is sufficient to rescue stress-triggered epithelial injury. |
| Institute: | China Pharmaceutical University |
| Department: | School of Medicine |
| Laboratory: | metabonomics |
| Last Name: | Yunalong |
| First Name: | Hou |
| Address: | Nanjing, Jiangsu |
| Email: | jian2103@163.com |
| Phone: | 18851105337 |
Subject:
| Subject ID: | SU001699 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Female |
Factors:
Subject type: Cultured cells; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA137921 | 13C Fru1 | 13C Fructose |
| SA137922 | 13C Fru3 | 13C Fructose |
| SA137923 | 13C Fru2 | 13C Fructose |
| SA137924 | Fru3 | Fructose |
| SA137925 | Fru1 | Fructose |
| SA137926 | Fru2 | Fructose |
| Showing results 1 to 6 of 6 |
Collection:
| Collection ID: | CO001692 |
| Collection Summary: | Isolation and Culture of Mouse Intestinal Organoids |
| Sample Type: | Intestine |
Treatment:
| Treatment ID: | TR001712 |
| Treatment Summary: | For metabolic flux analysis of 13C-labeled glucose, intestinal organoids were pretreated with unlabeled fructose (10 mM) for 24 hr, washed with PBS, and then switched into 13C labeled glucose-containing DMEM/F12 medium (10 mM U-[13C] D-glucose, Cambridge Isotope Laboratories) for 6 hr. To trace the metabolism of fructose, intestinal organoids were cultured in unlabeled fructose-containing DMEM/F12 medium (10 mM D-fructose, 0 mM D-glucose, 2.5 mM L-glutamine) for 12 hr, and then switched into 13C labeled fructose-containing medium (10 mM U-[13C] D-fructose, Sigma-aldrich) for 6 hr. |
Sample Preparation:
| Sampleprep ID: | SP001705 |
| Sampleprep Summary: | To harvest intracellular metabolites, organoid samples were washed with pre-cooled PBS for three times and metabolism was quenched with 80% ice-cold methanol. Pre-cooled methanol (1mL) with para-chlorophenylalanine (1 µM) as the internal standard was added to each sample and incubated on ice for 10 min. Sample lysates were then centrifuged at 4 °C for 15 min at 23,000 g. |
Combined analysis:
| Analysis ID | AN002657 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | UPLC-Q-TOF-MS (Synapt G2si, waters) |
| Column | Waters XBridge Amide (100 x 4.6mm,3.5um) |
| MS Type | ESI |
| MS instrument type | GC-TOF |
| MS instrument name | Waters Synapt-G2-Si |
| Ion Mode | NEGATIVE |
| Units | abundance |
Chromatography:
| Chromatography ID: | CH001960 |
| Instrument Name: | UPLC-Q-TOF-MS (Synapt G2si, waters) |
| Column Name: | Waters XBridge Amide (100 x 4.6mm,3.5um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002464 |
| Analysis ID: | AN002657 |
| Instrument Name: | Waters Synapt-G2-Si |
| Instrument Type: | GC-TOF |
| MS Type: | ESI |
| MS Comments: | MS MassLynx v4.1 Quanlynx v4.1 |
| Ion Mode: | NEGATIVE |
