Disease-related metabolites in culture medium of fibroblasts from patients with D-2-hydroxyglutaric aciduria, L-2-hydroxyglutaric aciduria, and combined D/L-2-hydroxyglutaric aciduria

Clin Chem. 2003 Jul;49(7):1133-8. doi: 10.1373/49.7.1133.


Background: D-2-Hydroxyglutaric aciduria (D-2-HGA), L-2-hydroxyglutaric aciduria (L-2-HGA), and the combined D/L-2-hydroxyglutaric aciduria (D/L-2-HGA) are poorly understood organic acidurias. To investigate the usefulness of cultured human skin fibroblasts for both diagnostic and research purposes, we measured disease-related metabolites in the cell culture medium.

Methods: We measured D-2-hydroxyglutarate (D-2-HG), L-2-hydroxyglutarate (L-2-HG), succinate, 2-ketoglutarate, and citrate in fibroblast cell medium by stable-isotope-dilution gas chromatography-mass spectrometry and glutamine, glutamic acid, and lysine with an amino acid analyzer. We used six cell lines from patients with D-2-HGA, two from patients with L-2-HGA, three from patients with D/L-2-HGA, and seven control cell lines. Culture medium was analyzed after a 96-h incubation period.

Results: Culture media from cell lines from D-2-HGA patients contained D-2-HG at concentrations 5- to 30-fold higher than media from controls, whereas the concentration of L-2-HG in media was not increased. Media from L-2-HGA cell lines showed a fivefold increase in L-2-HG compared with controls. Media containing fibroblasts from D/L-2-HGA patients contained moderately increased amounts of both D-2-HG and L-2-HG. For all cell lines, succinate concentrations in the blank medium were higher than after 96 h of incubation with the exception of two of three D/L-2-HGA cell lines. Media of D-2-HGA cell lines had 2-ketoglutarate concentrations that were 40% of that for controls. Glutamic acid concentrations in media of these cell lines were 60% lower than in controls.

Conclusions: Cell culture media from fibroblasts from patients with D-2-HGA, L-2-HGA, or D/L-2-HGA contain increased amounts the corresponding 2-HGs, demonstrating the suitability of fibroblasts for both diagnosis of and research concerning 2-HGAs.

MeSH terms

  • Biomarkers / analysis
  • Cell Line
  • Culture Media
  • Fibroblasts / metabolism*
  • Fibroblasts / pathology
  • Gas Chromatography-Mass Spectrometry
  • Glutamic Acid / metabolism
  • Glutamine / metabolism
  • Glutarates / chemistry
  • Glutarates / metabolism*
  • Glutarates / urine*
  • Humans
  • Ketoglutaric Acids / metabolism
  • Lysine / metabolism
  • Metabolic Diseases / metabolism*
  • Metabolic Diseases / pathology
  • Stereoisomerism
  • Succinic Acid / metabolism


  • Biomarkers
  • Culture Media
  • Glutarates
  • Ketoglutaric Acids
  • Glutamine
  • alpha-hydroxyglutarate
  • Glutamic Acid
  • Succinic Acid
  • Lysine