Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA064318

Local Sample ID:	Sample # 2
Subject ID:	SU001062
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Treatment:

Treatment ID:	TR001076
Treatment Summary:	Metabolic profiling will be conducted similarly to published methods and standard methods from our metabolic core. Glucose, glutamine and lactate levels in culture medium will be measured using gas chromatography/mass spectroscopy (GC/MS). Briefly, cells will be seeded in 6 well plates in triplicate using our standard neurosphere media (MH+++) and cultured for 24 to 48h before experiments. Changes in metabolite concentrations relative to fresh media will be normalized to protein content of each well. Cellular metabolite levels will be measured using standard protocols using deuterated 2-hydroxyglutarate (d5-5HG) as an internal standard and analyzed using GC/MS. To determine the fraction of metabolites from Glu and Gln, 13C versions of each metabolite [U-13C]glucose or [U-13C]glutamine (Cambridge Isotope Lab) will be used. All of these experiements will be performed by the Mayo Clinic Metabolomics Research Core.

Treatment ID:

TR001076

Treatment Summary:

Metabolic profiling will be conducted similarly to published methods and standard methods from our metabolic core. Glucose, glutamine and lactate levels in culture medium will be measured using gas chromatography/mass spectroscopy (GC/MS). Briefly, cells will be seeded in 6 well plates in triplicate using our standard neurosphere media (MH+++) and cultured for 24 to 48h before experiments. Changes in metabolite concentrations relative to fresh media will be normalized to protein content of each well. Cellular metabolite levels will be measured using standard protocols using deuterated 2-hydroxyglutarate (d5-5HG) as an internal standard and analyzed using GC/MS. To determine the fraction of metabolites from Glu and Gln, 13C versions of each metabolite [U-13C]glucose or [U-13C]glutamine (Cambridge Isotope Lab) will be used. All of these experiements will be performed by the Mayo Clinic Metabolomics Research Core.