DataEntryID 58
    General information
      Manuscript title: Dynamics and control of the central carbon metabolism in Hepatoma cells.
      PubMed ID: http://www.ncbi.nlm.nih.gov/pubmed/20426867
      Journal: BMC Systems Biology
      Year: 2010
      Authors: Klaus Maier, Ute Hofmann, Mathias Reuss and Klaus Mauch
      Affiliations: Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany
      Keywords: Hepatoma Cells, in vivo Dynamics, Dynamic Model, Control
      Full text article: https://kimosys.org/rails/active_storage/blobs/eyJfcmFpbHMiOnsibWVzc2FnZSI6IkJBaHBBbmdFIiwiZXhwIjpudWxsLCJwdXIiOiJibG9iX2lkIn19--d6c64c2fef47bbbe32782b68d2366188d6d3f4b8/Maier_2010.pdf
      Project name: not specified

    Experiment description
      Organism: Homo sapiens
      Strain: HepG2 (ATCC number HB-8065)
      Data type: metabolites at steady-state
      Data units: (mmol/L)
      Execution date: not specified

    Experimental details
      Temperature (°C): 37.0
      pH: not specified
      Carbon source: glucose
      Culture mode: batch
      Process condition: aerobic
      Dilution rate (h⁻¹): —
      Working volume: not specified L
      Biomass concentration (g/L): not specified
      Medium composition: Alanyl-glutamine-free William's medium E (PAN Biotech GmbH) that was supplemented with penicillin (100 U/mL), streptomycin (100 mg/mL), and Gibco Insulin-Transferrin-Selenium (100X) supplement (Invitrogen, Karlsruhe, Germany).
      General protocol information: Sampling Method: Removal of cell culture medium by suction (5 seconds).; Quenching: hot air; Extraction list: hot water; Analysis list: GC-MS, LC-MS, HPLC-UV/RI;
      Methods description: Extra- and intracellular samples were collected in triplicate directly before and after the stimulus, as well as 1, 2, 5, 10, 30, 60, 120, and 180 min after glucose deprivation. The sampling approach and the processing of the samples were done as previously described [1].
The concentrations of Lanine, serine, glucose, lactate, pyruvate, fumarate, malate, cis-aconitate, isocitrate, and citrate were determined by GC-MS as described before [1,2]. After glucose deprivation, extracellular glucose concentrations were determined in 10 μl of diluted (1:9 v/v) medium samples, the intracellular glucose concentrations before and after perturbation were determined in 5 and 50 μl of cell extract, respectively. Phosphoenolpyruvate, 3-phosphoglycerate, dihydroxyacetonphosphate, fructose-1,6-bisphosphate, glucose-6-phosphate, 6-phosphogluconate, sedoheptulose-7-phosphate, ribose-5-phosphate, and ribulose-5-phosphate were determined by LC-MS-MS as described by Hofmann [1] with the following modifications: HPLC separation was performed at 20 °C on a Synergi Hydro-RP column (150 × 2 mm, 4 μm; Phenomenex, Aschaffenburg, Germany) at a flow rate of 0.2 ml/min. Nucleotide analysis was performed by reversed phase ion pair high performance liquid chromatography. The HPLC system (Agilent Technologies, Waldbronn, Germany) consisted of an Agilent 1200 series autosampler, an Agilent 1200 series Binary Pump SL, an Agilent 1200 series thermostatted column compartment, and an Agilent 1200 series diode array detector set at 260 and 340 nm. The nucleotides were separated and quantified on an RP-C-18 column that was combined with a guard column (Supelcosil LC-18-T; 15 cm × 4.6 mm, 3 μm packing and Supelguard LC-18-T replacement cartridges, 2 cm; Supelco, Bellefonte, USA) at a flow rate of 1 ml/min.

-----------References-----------
[1] Hofmann U, Maier K, Niebel A, Vacun G, Reuss M, Mauch K. Identification of metabolic fluxes in hepatic cells from transient 13C-labeling experiments: Part I. Experimental observations. Biotechnol Bioeng 2008, 100:344-354. http://doi.org/chtgpd
[2] Maier K, Hofmann U, Bauer A, Niebel A, Vacun G, Reuss M, Mauch K. Quantification of statin effects on hepatic cholesterol synthesis by transient (13)C-flux analysis. Metab Eng 2009, 11:292-309. http://doi.org/dbkj9b 
      Data file: http://kimosys.org/repository/58/download?parameter=1143; 
      Alternative formats: http://kimosys.org/repository/58/attached_files/127/download; http://kimosys.org/repository/58/attached_files/46/download; 

    Submission and curation
      Entered by: Administrator KiMoSys
      Created: 2013-06-06 13:47:09 UTC
      Updated: 2020-04-24 16:10:32 UTC
      Version: 0
      Status: (reviewed) 2013-12-06 17:17:38 UTC
      Views: 314
      Downloads: 50