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MB Sample ID: SA017207
Local Sample ID: | 5Glucose10minReplicate1_0068.d |
Subject ID: | SU000401 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
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Collection:
Collection ID: | CO000395 |
Collection Summary: | HepG2 cells (ATCC HB-8065) were cultured in MEM containing 10 % (v:v) FBS, 100 U/mL penicillin and 100 mg/mL streptomycin (Invitrogen, Carlsbad, CA), 1 x MEM non-essential amino acids, and 5.5 mM glucose at 37 °C in a 5 % CO2 environment. Cells were grown for four to six passages in 10 cm tissue culture dishes with 14 mL MEM, and transferred to MULTIWELL™ 12 well culture dishes for incubation in 2 mL of the treatment medium. Upon reaching 80 % confluency, the cell culture medium was changed to media representative of experimental condition: 5.5 mM glucose MEM, 5.5 mM glucose MEM + 5 mM glucose, or 5.5 mM glucose MEM + 5 mM fructose. Media was replenished after 24 and 48 h. Cell material was collected at 10 min, 1, 6, and 24 h time points following the 48 h media replacement. Culture dishes were placed on ice and each well was washed twice with 1 mL ice-cold PBS. 4 mL ice-cold 3:1 methanol/H2O extraction solvent was added to each well. Cell material was manually scraped from each well, and the extraction solvent cell material suspension was transferred to collection tubes and frozen at −80 °C prior to further processing. |
Collection Protocol Filename: | metabolomic_responses_of_cultured_HepG2_liver_cells.pdf |
Sample Type: | Cell |
Collection Location: | Invitrogen, Carlsbad, CA |
Collection Frequency: | Cell material was collected at 10 min, 1, 6, and 24 h time points following the 48 h media replacement. |
Tissue Cell Identification: | HepG2 |