title: De novo biosynthesis of glycosaminoglycans in the extracellular matrix of skin studied by matrix-assisted laser desorption/ionization mass spectrometry.
abstract: The self-healing capacity of skin is limited, and medical intervention is often unavoidable. Skin may be generated ex vivo from cultured fibroblasts. Because the molecular composition of de novo formed skin (mostly collagen and glycosaminoglycans [GAGs]) is crucial, analytical methods are required for the quality control of tissue-engineered products. Here, we show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of fibroblast cultures subsequent to digestion with chondroitinase ABC is a reliable and fast method to monitor the GAG content of native and bioengineered skin. Furthermore, the supplementation of the fibroblast medium with ¹³C-labeled glucose provides insights into the biosynthesis of GAGs.
authors.affiliation: Institute of Medical Physics and Biophysics, Faculty of Medicine, University of Leipzig, D-04107 Leipzig, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Böhme
authors.forename: Ulf
authors.initials: U
authors.lastname: Anderegg
authors.forename: Ariane
authors.initials: A
authors.lastname: Nimptsch
authors.forename: Kathrin
authors.initials: K
authors.lastname: Nimptsch
authors.forename: Michael
authors.initials: M
authors.lastname: Hacker
authors.forename: Michaela
authors.initials: M
authors.lastname: Schulz-Siegmund
authors.forename: Daniel
authors.initials: D
authors.lastname: Huster
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schiller
mesh_terms: D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000032:analysis*; D003470:Culture Media; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology*; D005947:Glucose / Q000737:chemistry; D006025:Glycosaminoglycans / Q000096:biosynthesis*; D006801:Humans; D012867:Skin / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D003470:Culture Media; D006025:Glycosaminoglycans; D019765:Chondroitin ABC Lyase; D005947:Glucose