Abstract
The proteome of a cell is a molecular fingerprint directly relating to the gene expression snapshot profile at a certain point of time or developmental stage. Monitoring the expansion and the differentiation state of stem cells by proteomic means seems therefore a very attractive method for diagnostic as well as for therapeutic purposes. We have investigated the protein expression patterns of umbilical cord blood-derived CD34+/AC133+ cells in order to obtain a most comprehensive view of the stem cell proteome. For this purpose, we have applied 2-D gel electrophoresis and 2-D chromatography for most efficient protein/peptide separation and characterisation. The proteins were identified after tryptic digestion by nano-HPLC coupled directly to an ion trap mass spectrometer. An extensive bioinformatic analysis of the protein obtained revealed a dynamic stem cell proteome. This means that the heterogeneity of protein expression patterns obtained from different stem cell preparations refers to a limited set of stem cell-specific house keeping proteins as well as to a large number of proteins which depend on (marginal) stimuli from the environment. Since those are difficult to standardise, snapshot views of the stem cell proteome reflect not only stem cell-intrinsic metabolism but also the strong influence of the sample history on protein expression patterns.
Keywords: Stem cells, Umbilical cord blood, 2-D Liquid chromatography, 2-D Gel electrophoresis, Mass spectrometry
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