novo biosynthesis of glycosaminoglycans in the extracellular matrix of skin studied by matrix-assisted laser desorption/ionization mass spectrometry

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 f...

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Bibliographic Details
Published in:Analytical biochemistry 2012, Vol.421 (2), p.791-793
Main Authors: Böhme, Julia, Anderegg, Ulf, Nimptsch, Ariane, Nimptsch, Kathrin, Hacker, Michael, Schulz-Siegmund, Michaela, Huster, Daniel, Schiller, Jürgen
Format: Article
Language:English
Subjects:
biosynthesis
carbon
collagen
desorption
digestion
extracellular matrix
fibroblasts
glucose
glycosaminoglycans
ionization
isotope labeling
matrix-assisted laser desorption-ionization mass spectrometry
medical treatment
quality control
stable isotopes
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Summary: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.
ISSN:0003-2697
1096-0309