Mass Spectrometry raws files-metabololites from the secretome of Astrocytes (Progressive Supranuclear Palsy patients and controls)
Raw files from mass spectrometry (MS) for metabolomic analysis of the secretome from astrocytes differentiated from iPSC (reprogramed fibroblasts) of Progressive Supranuclear Palsy patients (PK01, PK02, PK04, PK08, PK09) and controls (PK03, PK05, PK06, CQ16) in triplicates.
Steps to reproduce
Astrocytes were cultured (in triplicates) in an astrocyte differentiation medium (AGM, Lonza #3186) for approximately 5 weeks until passage 5. The secretome was collected from astrocytes in passage 5. Astrocyte medium was switched to in DMEM/F12 without phenol red (Gibco # 21041-025) without fetal bovine serum (FBS) for 48h. The supernatants were collected and concentrated in 3000 KDa Ultracel® filters (Merck #UFC900324) and frozen at -80°C until processing form mass spectrometry (MS) analysis. Each sample was prepared using 20 µL of astrocyte supernatant, mixed with 200 µL of tetrahydrofuran (THF), followed by vortex for 30 seconds and the addition of 780 µL of methanol. The resulting solution was vortexed until homogenization for 30 seconds and the supernatant was separated by centrifugation at 3200 rpm, for 5 min at 4°C. Twenty microliters of the supernatant were diluted in 480 µL of methanol. One microliter of formic acid was added to the final aliquot for analysis in positive ion mode through direct infusion in an ESI-LTQ-XL Discovery (Thermo Scientific, Bremen, Germany) mass spectrometer. The equipment setup was as follows: flow rate of 10 μL.min−1, capillary temperature at 275 °C, spray voltage of 5 kV, and nitrogen sheath gas at 8 arbitrary units. A mass range of 150-1500 m/z was used for spectrum data acquisition. Experimental groups Progressive Supranuclear Palsy patients (PK01, PK02, PK04, PK08, PK09) controls (PK03, PK05, PK06, CQ16)
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