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MB Sample ID: SA205754
Local Sample ID: | OI1_D1_S1_Medium |
Subject ID: | SU002228 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002228 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
OI1_D1_S1_Medium | SA205754 | FL025242 | D1 | Experiment day |
Collection:
Collection ID: | CO002221 |
Collection Summary: | hAMSCs conditioned media samples were collected in triplicate at days 1, 6, 9, 12, 14, 16, and 21 after filtration through 40 μm pore strainers. |
Sample Type: | Stem cells |
Treatment:
Treatment ID: | TR002240 |
Treatment Summary: | hAMSCs were detached, at passage 7, from the flasks by trypsinization, counted in a Neubauer chamber and seeded at a density of 0.5 × 10 6 cells/flask. Cells were maintained under basal conditions until reaching ~100% confluence, then the basal culture medium was exchanged and supplemented with osteoinductive factors, specifically 10 mM β-glycerophosphate (β-GP, Sigma-Aldrich G9422), 50 µg/mL L-ascorbic acid (Sigma A0278), and 10 nM Dexa (ACROS Organics™ 230300010). |
Sample Preparation:
Sampleprep ID: | SP002234 |
Sampleprep Summary: | Both blank and conditioned media samples were subjected to a protein-precipitation procedure and dried under vacuum. Prior to NMR analysis, dried media samples were resuspended in 100 mM phosphate buffer at pH 7.4 (prepared in D2O) containing 0.1 mM 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid (TSP in D2O, Sigma-Aldrich 293040), homogenised and transferred to 5 mm NMR tubes. |
Extract Storage: | -80℃ |
Sample Resuspension: | 100 mM phosphate buffer at pH 7.4 (prepared in D2O) containing 0.1 mM 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid (TSP in D2O, Sigma-Aldrich 293040) |
Analysis:
MB Sample ID: | SA205754 |
Analysis ID: | AN003508 |
Analysis Type: | NMR |
Software Version: | Topspin 4.0.8 |
Operator Name: | Daniela Bispo |
Results File: | ST002143_AN003508_Results.txt |
Units: | ppm |
NMR:
NMR ID: | NM000238 |
Analysis ID: | AN003508 |
Instrument Name: | Bruker Avance III |
Instrument Type: | FT-NMR |
NMR Experiment Type: | 1D-1H |
Field Frequency Lock: | D2O |
Spectrometer Frequency: | 500 MHz |
NMR Probe: | TXI |
NMR Solvent: | 100 mM phosphate buffer at pH 7.4 (prepared in D2O) containing 0.1 mM 3-(trimethylsilyl)-propionic-2,2,3,3-d4 acid |
NMR Tube Size: | 5 mm |
Shimming Method: | Topshim |
Receiver Gain: | 203 |
Temperature: | 298 K |
Number Of Scans: | 256 |
Dummy Scans: | 4 |
Acquisition Time: | 2.3 s |
Relaxation Delay: | 4 s |
Spectral Width: | 7002.801 Hz |
Num Data Points Acquired: | 32k |
Zero Filling: | 131k |
Baseline Correction Method: | Manual |
Chemical Shift Ref Std: | TSP |