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MB Sample ID: SA338824
| Local Sample ID: | FLi44.3 |
| Subject ID: | SU003242 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003242 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| FLi44.3 | SA338824 | FL040115 | Fetal liver | Sample source |
| FLi44.3 | SA338824 | FL040115 | lps | exposure |
| FLi44.3 | SA338824 | FL040115 | 12 | timepoint |
Collection:
| Collection ID: | CO003235 |
| Collection Summary: | At 2, 5, 12 and 24h, dams were terminally anesthetized by subcutaneous injection of 0.2 ml of Zoletil mixture (tiletamin/zolazepam, xylazin og fentanyl) and killed by exanguination by withdrawal of heart blood into Eppendorf tubes containing 36 ml K2EDTA (N=7-9 per exposure/time point). The uterus was excised and opened. Fetuses were excised from their embryonic sac, their viability confirmed, killed by decapitation, sexed by visual inspection, and their position in the uterus noted. From each litter, the first female fetus encountered in the right uterine horn, counting from the cervix, was selected and saved for analyses. The placenta was dissected into chorion (chorionic plate, labyrinth and junctional zones) and decidua by blunt/stump dissection under stereomicroscope (Wild Heerbrugg, Switzerland)76. From dams, the liver and right lung were dissected. Dissected organs were snap frozen in liquid nitrogen and kept at -80ºC |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR003251 |
| Treatment Summary: | Lipopolysaccharide (LPS; E. Coli serotype 00:55 B5 LPS (Sigma Lot nr. 025M4040V)) was diluted to the final concentration (0.02 µg/µl) in double distilled pyrogen-free water (Chem-Lab, Zedelgem, Belgium). In the morning of GD 17, the pregnant mice were semi-randomized into control and LPS treatment groups (denoted Ctrl and LPS, respectively), evenly distributing weights among the groups. Out of 80 mice in total, 74 were pregnant. Animals were placed in a whole-body inhalation chamber with an attached anaesthetic vaporizer (Penlon Sigma Delta, Abingdon, UK), delivering 3-4% isoflurane in filtered air, and were intratracheally instilled with 50 µl of vehicle (Ctrl) or 1 μg LPS in 50 µl vehicle, followed by 200μl of air. Vehicle and LPS were administered through a 0.58 mm polyethylene tube (Ref: 427411, Becton Dickinson, Brøndby, Denmark) attached to a plastic syringe. The procedure has been shown not to affect gestation, offspring viability nor growth75. After instillation, animals were returned to their cage, briefly placed on heating pads and checked upon regularly until euthanization |
Sample Preparation:
| Sampleprep ID: | SP003249 |
| Sampleprep Summary: | Lipids were extracted from fetal liver samples (20 mg) using Folch extraction* with 8-12 replicates from each experimental group at each time point. Prior to tissue lysis, Splash mix (Merck) was added to the extraction solvent and tissue samples were lysed by beat beating in a FastPrep-24 homogenizer. After centrifugation and phase separation, the apolar and polar phases were transferred to separate tubes, and the apolar phase dried under N2. *Folch, J., Lees, M. & Sloane Stanley, G. H. A simple method for the isolation and purification of total lipides from animal tissues. J. Biol. Chem. 226, 497–509 (1957). |
Combined analysis:
| Analysis ID | AN005123 | AN005124 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Bruker Tims TOF flex | Bruker Tims TOF flex |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Peak intensity | Peak intensity |
Chromatography:
| Chromatography ID: | CH003877 |
| Chromatography Summary: | Reverse phase (C18), 10 min |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 55 |
| Flow Gradient: | from 0 to 0.5 min, 40–43% B; from 0.5 to 0.7 min, 43‐65% B; from 0.7 to 0.8 min, 65-70% B; from 0.8 to 2.3 min, 70-99% B; from 2.3 to 6 min, 99% B; from 6-6.8 min, 99-40% B; from 6.8-7 min before equilibration for 3 min with the initial conditions |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 60% acetonitrile/40% water; 10 mM ammonium formate; 0.1% formic acid |
| Solvent B: | 90% isopropanol/10% acetonitrile; 10 mM ammonium formate; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004859 |
| Analysis ID: | AN005123 |
| Instrument Name: | Bruker Tims TOF flex |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Data was acquired with Trapped ion mobility spectrometry (TIMS) activated. Bruker Metaboscape software was used for data processing. Annotation was done using the built in "lipid search" module and Lipid blast. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004860 |
| Analysis ID: | AN005124 |
| Instrument Name: | Bruker Tims TOF flex |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Data was acquired with Trapped ion mobility spectrometry (TIMS) activated. Bruker Metaboscape software was used for data processing. Annotation was done using the built in "lipid search" module and Lipid blast. |
| Ion Mode: | NEGATIVE |
