Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA069341

Local Sample ID:	ms6982-52
Subject ID:	SU001075
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Treatment:

Treatment ID:	TR001089
Treatment Summary:	Murine derived pure cortical astrocytes were plated at 4x10^6 density in 10cm petri dish and grown in astro growth medium (AGM) for 2-3 days until it reaches the 90% confluency. Day before the treatment with glutamine or glutamate AGM was changed to serum free “E” media. In case of isotomer analysis 40% of glucose was switched with U13C6 (glucose) and added to serum free “E” media. On the day of treatment first the spent media was collected and then washed with base media. After washing, astrocytes were exposed to different ratio of glutamate: glutamine for 1h under non-CO2 condition. 1h spent media was collected. After washing three times with phosphate buffer saline cell pellet was collected in ice cold 100% methanol by scrapping. All the samples were immediately transferred to -70C for future analysis. E media composition: DMEM (w/o phenol red)+ 1% PenStrep+ 10% FBS+ Glutamate+ Glucose Abbreviation: Gn-Glutamine Gt-Glutamate B- Base media G- Glucose P- Sodium pyruvate

Treatment ID:

TR001089

Treatment Summary:

Murine derived pure cortical astrocytes were plated at 4x10^6 density in 10cm petri dish and grown in astro growth medium (AGM) for 2-3 days until it reaches the 90% confluency. Day before the treatment with glutamine or glutamate AGM was changed to serum free “E” media. In case of isotomer analysis 40% of glucose was switched with U13C6 (glucose) and added to serum free “E” media. On the day of treatment first the spent media was collected and then washed with base media. After washing, astrocytes were exposed to different ratio of glutamate: glutamine for 1h under non-CO2 condition. 1h spent media was collected. After washing three times with phosphate buffer saline cell pellet was collected in ice cold 100% methanol by scrapping. All the samples were immediately transferred to -70C for future analysis. E media composition: DMEM (w/o phenol red)+ 1% PenStrep+ 10% FBS+ Glutamate+ Glucose Abbreviation: Gn-Glutamine Gt-Glutamate B- Base media G- Glucose P- Sodium pyruvate