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MB Sample ID: SA205747
| Local Sample ID: | Medium_Blank_S3 |
| 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 |
|---|---|---|---|---|
| Medium_Blank_S3 | SA205747 | FL025240 | Blank | 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: | SA205747 |
| 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 |
