Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA023070

Local Sample ID:	M_12
Subject ID:	SU000476
Subject Type:	Human Cell Media
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

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Subject:

Subject ID:	SU000476
Subject Type:	Human Cell Media
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
M_12	SA023070	FL005612	DMSO	Treatment
M_12	SA023070	FL005612	Normal	Growth_type

Collection:

Collection ID:	CO000470
Collection Summary:	An aliquot of the media following treatment was collected, and the remainder of the media was aspirated.
Sample Type:	Fibroblasts

Treatment:

Treatment ID:	TR000490
Treatment Summary:	Primary cell cultures of desmoid tumor samples were established as previously described [1,2]. Monolayer cell cultures were grown in DMEM supplemented with 5% fetal bovine serum and maintained at 37°C in 5% CO2. Cells were divided when confluent and experiments were performed between the third and sixth passages. Approximately 10 x 106 cells were treated with 1.0uM Dasatinib (Selleck, Houston, USA) dissolved in DMSO, or 0.5uM FAK Inhibitor 14 (Cayman Chemicals Company, Michigan, USA) dissolved in water. Cells were incubated in fresh media containing the inhibitors, or vehicle, for 24 hours.

Treatment ID:

TR000490

Treatment Summary:

Primary cell cultures of desmoid tumor samples were established as previously described [1,2]. Monolayer cell cultures were grown in DMEM supplemented with 5% fetal bovine serum and maintained at 37°C in 5% CO2. Cells were divided when confluent and experiments were performed between the third and sixth passages. Approximately 10 x 106 cells were treated with 1.0uM Dasatinib (Selleck, Houston, USA) dissolved in DMSO, or 0.5uM FAK Inhibitor 14 (Cayman Chemicals Company, Michigan, USA) dissolved in water. Cells were incubated in fresh media containing the inhibitors, or vehicle, for 24 hours.

Sample Preparation:

Sampleprep ID:	SP000483
Sampleprep Summary:	Cell media was extracted with 50% acetonitrile:water, vortexed, and centrifuged. An analytical quality control (QC) total pool was generated from equal aliquots from all of the media samples, divided into three total pool aliquots, and processed identically to the media study samples. All study and pool samples were lyophilized to dryness and reconstituted in a 0.2M phosphate buffer, pH 7.4, in D2O with 10% Chenomx ISTD and 6mM Imidazole.

Sampleprep ID:

SP000483

Sampleprep Summary:

Cell media was extracted with 50% acetonitrile:water, vortexed, and centrifuged. An analytical quality control (QC) total pool was generated from equal aliquots from all of the media samples, divided into three total pool aliquots, and processed identically to the media study samples. All study and pool samples were lyophilized to dryness and reconstituted in a 0.2M phosphate buffer, pH 7.4, in D2O with 10% Chenomx ISTD and 6mM Imidazole.

Analysis:

MB Sample ID:	SA023070
Analysis ID:	AN000712
Analysis Type:	NMR
Num Factors:	14

NMR:

NMR ID:	NM000079
Analysis ID:	AN000712
Instrument Name:	Bruker Avance III
Instrument Type:	FT-NMR
NMR Experiment Type:	1D 1H
Spectrometer Frequency:	700 MHz