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Profiling and semiquantitative analysis of the cell surface proteome in human mesenchymal stem cells.

Authors
Lee, SK; Kim, JH; Kim, SS; Kang, T; Park, NH; Kwon, KH; Lee, SS; Lee, ZW; Suh-Kim, H; Cho, K; Yun, SY; Han, JY; Yoo, JS; An, HJ; Park, YM
Citation
Analytical and bioanalytical chemistry, 405(16):5501-5517, 2013
Journal Title
Analytical and bioanalytical chemistry
ISSN
1618-26421618-2650
Abstract
Mulitpotent mesenchymal stem cells (MSCs) derived from human bone marrow are promising candidates for the development of cell therapeutic strategies. MSC surface protein profiles provide novel biological knowledge concerning the proliferation and differentiation of these cells, including the potential for identifying therapeutic targets. Basic fibroblast growth factor (bFGF) affects cell surface proteins, which are associated with increased growth rate, differentiation potential, as well as morphological changes of MSCs in vitro. Cell surface proteins were isolated using a biotinylation-mediated method and identified using a combination of one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry. The resulting gel lines were cut into 20 bands and digested with trypsin. Each tryptic fragment was analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry. Proteins were identified using the Mascot search program and the International Protein Index human database. Noble MSC surface proteins (n = 1,001) were identified from cells cultured either with (n = 857) or without (n = 667) bFGF-containing medium in three independent experiments. The proteins were classified using FatiGO to elucidate their function. We also confirmed the proteomics results using Western blotting and immunofluorescence microscopic analysis. The nature of the proteins identified makes it clear that MSCs express a wide variety of signaling molecules, including those related to cell differentiation. Among the latter proteins, four Ras-related Rab proteins, laminin-R, and three 14-3-3 proteins that were fractionated from MSCs cultured on bFGF-containing medium are implicated in bFGF-induced signal transduction of MSCs. Consequently, these finding provide insight into the understanding of the surface proteome of human MSCs.
MeSH terms
14-3-3 Proteins/analysis/metabolismCell DifferentiationCells, CulturedElectrophoresis, Polyacrylamide GelFibroblast Growth Factor 2/pharmacologyHumansMass Spectrometry/methodsMesenchymal Stromal Cells/cytology/drug effects/*metabolismMultipotent Stem Cells/metabolismProteins/*analysis/classification/metabolismProteome/*analysisProteomics/*methodsReceptors, Laminin/metabolismReproducibility of ResultsSpectrometry, Mass, Electrospray IonizationTandem Mass Spectrometry
DOI
10.1007/s00216-013-6969-z
PMID
23657447
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Anatomy
AJOU Authors
김, 성수서, 해영
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