title: Nonviability of cells with oxidative defects in galactose medium: a screening test for affected patient fibroblasts.
abstract: Diagnosis of respiratory chain defects in cultured skin fibroblasts is a difficult diagnostic procedure. We investigated the feasibility of using survival of skin fibroblasts in culture medium with galactose as the major carbon source as a method of quickly diagnosing cell lines that were compromised in oxidative metabolism. We found that cells from patients with most forms of cytochrome oxidase deficiency, cells with complex I deficiency, cells with multiple respiratory chain defects and cells with severe pyruvate dehydrogenase (PDH) complex deficiency failed to survive when subcultured into galactose (5 mM) medium. Cells from patients with Lebers hereditary optic neuropathy (LHON), Kearns-Sayre syndrome (KSS), myoclonus-epilepsy-lactic acidosis-stroke (MELAS), the hepatic form of cytochrome oxidase deficiency, and mild PDH complex deficiency survived well in galactose (5 mM)-containing medium. This could be used as a rapid screening test for skin fibroblasts with major oxidative defects.
authors.affiliation: Department of Biochemistry, University of Toronto, Ontario, Canada.
authors.forename: B H
authors.initials: BH
authors.lastname: Robinson
authors.forename: R
authors.initials: R
authors.lastname: Petrova-Benedict
authors.forename: J R
authors.initials: JR
authors.lastname: Buncic
authors.forename: D C
authors.initials: DC
authors.lastname: Wallace
mesh_terms: D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D030401:Cytochrome-c Oxidase Deficiency; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005690:Galactose / Q000494:pharmacology*; D006801:Humans; D007625:Kearns-Sayre Syndrome / Q000378:metabolism* / Q000473:pathology; D017241:MELAS Syndrome / Q000378:metabolism* / Q000473:pathology; D008928:Mitochondria / Q000378:metabolism; D015418:Optic Atrophies, Hereditary / Q000378:metabolism* / Q000473:pathology; D011767:Pyruvate Decarboxylase / Q000378:metabolism; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism; D015325:Pyruvate Dehydrogenase Complex Deficiency Disease / Q000378:metabolism* / Q000473:pathology; D012867:Skin / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology
chemical_list: D011768:Pyruvate Dehydrogenase Complex; D011767:Pyruvate Decarboxylase; D005690:Galactose