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MB Sample ID: SA005453
| Local Sample ID: | Media blank |
| Subject ID: | SU000115 |
| Subject Type: | Human cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | SKOV3ip1 |
| Subject Comments: | p80, p85, p86, p89 |
| Cell Passage Number: | p80, p85, p86, p89 |
| Cell Counts: | p80, p85, p86, p89 |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000115 |
| Subject Type: | Human cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | SKOV3ip1 |
| Subject Comments: | p80, p85, p86, p89 |
| Cell Passage Number: | p80, p85, p86, p89 |
| Cell Counts: | p80, p85, p86, p89 |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Media blank | SA005453 | FL001370 | Media blank | Sample Type |
| Media blank | SA005453 | FL001370 | 4 hours | Timepoint |
Collection:
| Collection ID: | CO000098 |
| Collection Summary: | Samples were collected at 18 hours post co-culture |
| Collection Protocol Filename: | Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Enpdfannabinoid_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf |
| Sample Type: | Media |
| Storage Conditions: | -80 C |
Treatment:
| Treatment ID: | TR000116 |
| Treatment Summary: | The study investigated the interaction between omental adipocytes and OvCa cells, as a follow up to preliminary data indicating this leads to reprograming of metabolic (especially lipid) profiles in both adipocytes and OvCa cells as ovarian cancer cells (OvCa) readily metastasize to the omental fat pad in the abdomen and stimulate the release of fatty acids. In order to mimic the interaction between OvCa and omental adipocytes during metastasis, a coculture system was used that employed OvCa cells and primary human adipocytes isolated from omentum. Human primary adipocytes were isolated from omental explants from patients undergoing surgery for benign conditions. After surgical removal, omental tissue was digested with collagenase I, and primary cultures of adipocytes were established, characterized, and incorporated into the co-culture. The primary adipocytes were isolated and co-cultured with the OvCa cell line Skov3ip1. In this current submission, the the samples will be collected at 4, 18 and 24 hour time points post co-culture to determine the time dependent effect on lipid mediators, including oxylipins and ceramides. The study results included in this DRCC submission were the 18 hour time point data for oxylipins and ceramides. |
| Treatment Protocol Filename: | Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Enpdfannabinoid_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf |
| Cell Storage: | -80 C |
Sample Preparation:
| Sampleprep ID: | SP000111 |
| Sampleprep Summary: | See sample prep protocol file |
| Sampleprep Protocol Filename: | Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Enpdfannabinoid_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf |
| Processing Storage Conditions: | - 20 °C |
| Extraction Method: | SPE |
| Extract Concentration Dilution: | 250 µL |
| Extract Cleanup: | SPE |
| Extract Storage: | - 20 °C |
| Sample Resuspension: | 100 µL |
| Sample Spiking: | See sample prep protocol file |
Combined analysis:
| Analysis ID | AN000152 | AN000153 | AN000154 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | |||
| Chromatography system | |||
| Column | |||
| MS Type | ESI | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex API 4000 QTrap | ABI Sciex API 4000 QTrap | ABI Sciex API 4000 QTrap |
| Ion Mode | NEGATIVE | POSITIVE | POSITIVE |
| Units | Area % | nM | nM |
Chromatography:
| Chromatography ID: | CH000108 |
| Chromatography Summary: | Oxylipin analysis |
| Methods Filename: | Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf |
| Column Temperature: | 60 C |
| Flow Gradient: | See protocol/methods file |
| Flow Rate: | 0.25 |
| Internal Standard: | See protocol/methods file |
| Retention Time: | See protocol/methods file |
| Sample Injection: | 5 L |
| Solvent A: | 100% water; 0.1% acetic acid |
| Solvent B: | 90% acetonitrile/ 10% isopropanol |
| Analytical Time: | 16 min |
| Weak Wash Solvent Name: | 20% methanol, 10% isopropanol |
| Weak Wash Volume: | 600 L |
| Strong Wash Solvent Name: | 50:50 Acetonitrile:Methanol |
| Strong Wash Volume: | 600 L |
| Sample Loop Size: | 17 L |
MS:
| MS ID: | MS000128 |
| Analysis ID: | AN000152 |
| Instrument Name: | ABI Sciex API 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Oxylipins analysis |
| Ion Mode: | NEGATIVE |
| Ion Source Temperature: | See protocol/methods file |
| Ion Spray Voltage: | See protocol/methods file |
| MS ID: | MS000129 |
| Analysis ID: | AN000153 |
| Instrument Name: | ABI Sciex API 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Ceramide analysis |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | See protocol/methods file |
| Ion Spray Voltage: | See protocol/methods file |
| MS ID: | MS000130 |
| Analysis ID: | AN000154 |
| Instrument Name: | ABI Sciex API 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Endocannabinoid analysis |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | See protocol/methods file |
| Ion Spray Voltage: | See protocol/methods file |
