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MB Sample ID: SA007398
| Local Sample ID: | SeMSC (PNT1A) |
| Subject ID: | SU000153 |
| Subject Type: | Human cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | ATCC, Sigma |
| Cell Strain Details: | DU145 prostate cancer, PNT1A prostate nontumor |
| Subject Comments: | 10 |
| Cell Primary Immortalized: | Immortalized |
| Cell Passage Number: | 10 |
| Cell Counts: | 10 |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000153 |
| Subject Type: | Human cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | ATCC, Sigma |
| Cell Strain Details: | DU145 prostate cancer, PNT1A prostate nontumor |
| Subject Comments: | 10 |
| Cell Primary Immortalized: | Immortalized |
| Cell Passage Number: | 10 |
| Cell Counts: | 10 |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| SeMSC (PNT1A) | SA007398 | FL001882 | SeMSC | Treatment |
| SeMSC (PNT1A) | SA007398 | FL001882 | PNT1A | Prostate Cell Line |
Collection:
| Collection ID: | CO000137 |
| Collection Summary: | After incubation for 24 hrs, cells were washed twice with 0.5 mL PBS in D2O (Cambridge Isotope Laboratories). D2O (0.7 mL) was again added to each petri dish and cells were gently scraped from the surface with a sterile cell scraper. A volume of 0.7 mL of the harvested cell suspension was transferred into a 5 mm NMR tube and 10 mL of 100 mM TMSP in D2O was added to each tube. |
| Sample Type: | cell line |
| Collection Method: | scraping cells from flask |
| Collection Location: | lab |
| Collection Frequency: | once a week |
| Volumeoramount Collected: | 0.7 mL per dish |
| Storage Conditions: | in ice |
| Collection Tube Temp: | in ice |
Treatment:
| Treatment ID: | TR000155 |
| Treatment Summary: | Cells were grown to 80% confluence in Petri dishes (50 mm in diameter) in 4.5% CO2 at 37 OC for 5-6 days for full attachment to the substratum. After aspirating the old growth medium, 3 mL of SeM or SeMSC (300 mM) in fresh pre-warmed medium at 37 ºC was added to each dish. |
| Treatment Compound: | Selenomethionine / Se-methylselenocysteine |
| Treatment Dose: | 300 mM |
| Treatment Dosevolume: | 3 mL |
| Treatment Doseduration: | 24 hrs |
| Treatment Vehicle: | dissolved in culture media |
| Cell Storage: | incubator |
| Cell Growth Container: | petri dish (Corning) |
| Cell Growth Config: | monolayer, adherent |
| Cell Growth Rate: | 5-6 days |
| Cell Inoc Proc: | trypsinized, then subculture |
| Cell Media: | RPMI, EMEM |
| Cell Envir Cond: | 37 C, 4.5% CO2 incubator |
| Cell Harvesting: | after trypsinization |
| Cell Pct Confluence: | 80% |
| Cell Media Lastchanged: | 2-3 days |
Sample Preparation:
| Sampleprep ID: | SP000150 |
| Sampleprep Summary: | Cells were grown to 80% confluence in Petri dishes (50 mm in diameter) in 4.5% CO2 at 37 OC for 5-6 days for full attachment to the substratum. After aspirating the old growth medium, 3 mL of SeM or SeMSC (300 mM) in fresh pre-warmed medium at 37 ºC was added to each dish. After incubation for 24 hrs, cells were washed twice with 0.5 mL PBS in D2O (Cambridge Isotope Laboratories). D2O (0.7 mL) was again added to each petri dish and cells were gently scraped from the surface with a sterile cell scraper. A volume of 0.7 mL of the harvested cell suspension was transferred into a 5 mm NMR tube and 10 mL of 100 mM TMSP in D2O was added to each tube. Prior to NMR analysis, tubes were vortexed to ensure samples were in suspension. Control experiments using untreated cells were conducted in parallel with treated cells with the same incubation protocols and sample preparation. |
| Sampleprep Protocol Filename: | Prostate_Cells_Metabolomics_Procedure.doc |
| Sample Resuspension: | in deuterated water |
| Cell Type: | prostate |
Analysis:
| MB Sample ID: | SA007398 |
| Analysis ID: | AN000216 |
| Laboratory Name: | Purdue Chemistry NMR Facility |
| Analysis Type: | NMR |
| Acquisition Date: | May 30, 2014; June 6, 2014 |
| Software Version: | AV-III-800 |
| Operator Name: | Dr. John Harwood |
| Detector Type: | TX1 |
| Chromatography ID: | CH000148 |
| Num Factors: | 4 |
| Num Metabolites: | 12 |
NMR:
| NMR ID: | NM000050 |
| Analysis ID: | AN000216 |
| Instrument Name: | Bruker Avance III |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D 1H |
| Field Frequency Lock: | Deuterium |
| Standard Concentration: | 1.43 mM |
| Spectrometer Frequency: | 800 MHz |
| NMR Probe: | TX1, inverse |
| NMR Solvent: | D2O |
| NMR Tube Size: | 5 mm x 7 in |
| Shimming Method: | gradient, topshim |
| Pulse Sequence: | zg one-pulse |
| Water Suppression: | presaturation |
| Pulse Width: | 8.5 msec |
| Power Level: | 12.25 W |
| Receiver Gain: | 912 |
| Offset Frequency: | 4.8 ppm |
| Presaturation Power Level: | 57 dB |
| Chemical Shift Ref Cpd: | TMSP |
| Temperature: | 295.9 |
| Number Of Scans: | 256 |
| Dummy Scans: | 8 |
| Acquisition Time: | 1.4680564 |
| Relaxation Delay: | 6.5 msec |
| Spectral Width: | 11160 Hz |
| Num Data Points Acquired: | 32 K |
| Real Data Points: | 32 K |
| Line Broadening: | 1 Hz |
| Zero Filling: | 16K |
| Apodization: | lorentzian |
| Baseline Correction Method: | polynomial |
| Chemical Shift Ref Std: | TMSP |
