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MB Sample ID: SA356298
Local Sample ID: | CD_PC9_3_1_CTL1_rep2 |
Subject ID: | SU003406 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
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Subject:
Subject ID: | SU003406 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
CD_PC9_3_1_CTL1_rep2 | SA356298 | FL041595 | Non-small cell lung cancer cells | Sample source |
CD_PC9_3_1_CTL1_rep2 | SA356298 | FL041595 | PC-9 | Cell line |
CD_PC9_3_1_CTL1_rep2 | SA356298 | FL041595 | Monoculture | Culture Type |
CD_PC9_3_1_CTL1_rep2 | SA356298 | FL041595 | No treatment | Immune checkpoint inhibitor |
Collection:
Collection ID: | CO003399 |
Collection Summary: | Using Nivolumab (anti-PD1) and Ipilimumab (anti-CTLA4), we pharmacologically modulated non-small cell lung cancer (NSCLC) cell lines in co-culture with CD8+ T lymphocytes (T CD8+) obtained from healthy blood donors. Two different modalities of co-cultures were established: direct and indirect (with T CD8+ separated from the cancer cells by a Boyden chamber with 1.0 μm pores). 1H-NMR spectroscopy was conducted to characterize the impact of the ICI in the metabolic profile of the immune and cancer cells, as well as the influence of immune cells in the cancer cells exometabolome. Therefore, the media obtained from the established cultures were centrifuged at 135 × g for 5 minutes and the supernatants were collected, stored at -80⁰C and later explored and defined by NMR. |
Sample Type: | Non-small cell lung cancer |
Treatment:
Treatment ID: | TR003415 |
Treatment Summary: | For the experimental conditions, cells were exposed to Nivolumab (10 mg/mL, OPDIVO), anti-PD1) or Ipilimumab (5 mg/mL, YERVOY, anti-CTLA-4) for 48 h. The Nivolumab and Ipilimumab concentrations were chosen in accordance to the posology indications from the European Medicines Agency (EMA) and using as reference a male adult with 80 kg and 5 L of blood. The conditions were tested in monoculture or in co-culture with TCD8+. |
Sample Preparation:
Sampleprep ID: | SP003413 |
Sampleprep Summary: | The supernatant samples were diluted in Milli-Q water to make up 540 μL (150 μL of water was added to 390 μL of supernatant). A volume of 30 µL of phosphate buffer was added to the samples along with 30 μL of a solution of 2.2 mM 3-(trimethylsilyl) propionic 2,2,3,3-d4 acid (TSP) in D2O. TSP was used as an internal reference for quantification and chemical shift. |
Analysis:
MB Sample ID: | SA356298 |
Analysis ID: | AN005383 |
Analysis Type: | NMR |
Num Factors: | 27 |
Num Metabolites: | 28 |
Units: | nanomoles |
NMR:
NMR ID: | NM000285 |
Analysis ID: | AN005383 |
Instrument Name: | Bruker Ultrashield 500 Plus |
Instrument Type: | FT-NMR |
NMR Experiment Type: | 1D-1H |
NMR Comments: | Spectra were acquired in a magnetic field of 500 MHz in an Ultrashied 500 Plus spectrometer (Bruker) with a TCI-Z probe at Instituto de Tecnologia Química e Biológica da Universidade NOVA de Lisboa (ITQB-UNL), Oeiras. The spectra analysis was made in TopSpin 4.1 software, according to Chenomx NMR suite 8.11 and Human Metabolome Database (HMDB). |
Spectrometer Frequency: | 500 MHz |
NMR Probe: | 5 mm prodigy TCI-z |
NMR Solvent: | 10% D2O/90% H20 |
NMR Tube Size: | 5 mm |