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MB Sample ID: SA125861
| Local Sample ID: | C1 |
| Subject ID: | SU001566 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
| Genotype Strain: | Fischer 344 |
| Age Or Age Range: | 6 wk |
| Gender: | Female |
| Animal Animal Supplier: | Envigo |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001566 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
| Genotype Strain: | Fischer 344 |
| Age Or Age Range: | 6 wk |
| Gender: | Female |
| Animal Animal Supplier: | Envigo |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| C1 | SA125861 | FL015450 | immobilized IFNy | Treatment group |
Collection:
| Collection ID: | CO001561 |
| Collection Summary: | Neural stem cells were collected from the subventricular zone of the brain in a 6-8 wk old rat. Papain tissue dissociation was used to isolate cells and the neurosphere culture system used to expand them for up to 7 passages. |
| Sample Type: | Brain |
Treatment:
| Treatment ID: | TR001581 |
| Treatment Summary: | Neurospheres were dissociated and plated onto laminin functionalized soft chitosan hydrogel surfaces. Three groups were grown for 7 days on their substrate in basal media . One group had no IFNy, another group 300 ng/mL soluble IFNy and the final group 300 ng/mL hydrogel immobilized IFNy. |
Sample Preparation:
| Sampleprep ID: | SP001574 |
| Sampleprep Summary: | NSC seeded gels were snap frozen and stored at -80C until extraction. Two gels were combined per group and extracted using a modified Bligh and Dyer extraction technique. In brief, 100 uL of methanol was added to each sample then underwent a series of snap freezing, sonication and vortexing three times. 750 μl of 1:2 chloroform:methanol was added to each homogenized sample, vortexed, then an additional 250 μL of chloroform was added, finally 250 μL water was added, all solvents used were LC grade. Samples were stored in -20 overnight and centrifuged to separate the phase layers and solidify the protein precipitate interface. Aqueous and organic layers were separated, dried down using Centrivap (Labconco) and stored in the -80 C until use. Aqueous portions of the extract were resuspended in 200 μL of 35% acetonitrile. |
Combined analysis:
| Analysis ID | AN002474 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Eksigent microLC 200 |
| Column | Phenomenex Luna NH2(150 x 1.0mm,3um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 5600+ TripleTOF |
| Ion Mode | POSITIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH001812 |
| Chromatography Summary: | The mobile phases for separation consisted of water (A) and acetonitrile (B), both supplemented with 5 mM ammonium acetate and adjusted to pH 7.3 using ammonium hydroxide. The gradient proceeded at a flow rate of 30 μL/min as follows: 98% B at 0 min, 95% B at 1 min, 80% B at 5 min, 46% B at 6 min, 14.7% B at 13 min, 0% B at 17 min, 100% B at 17.1 min, and 100% B at 23 min. |
| Chromatography Comments: | Phenomenex (Luna 3 μ NH2 100 Å, 150 mm × 1.0 mm) |
| Instrument Name: | Eksigent microLC 200 |
| Column Name: | Phenomenex Luna NH2(150 x 1.0mm,3um) |
| Solvent A: | 100% water |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002294 |
| Analysis ID: | AN002474 |
| Instrument Name: | ABI Sciex 5600+ TripleTOF |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Independent data acquisition was used with rolling collision energy for selected parent ions.Retention times and mass to charge values were aligned between samples using MarkerView software. Putative metabolite identifications were made using metaboanalyst, masstrix and HMDB softwares/databases. |
| Ion Mode: | POSITIVE |
