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

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 potenti...

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Bibliographic Details
Published in:Analytical and bioanalytical chemistry 2013-06, Vol.405 (16), p.5501-5517
Main Authors: Lee, Sang Kwang, Kim, Jae Ho, Kim, Sung-Soo, Kang, Taewook, Park, Nam Hyun, Kwon, Kyung-Hoon, Lee, Sang Sook, Lee, Zee Won, young Suh-Kim, Hae, Cho, Kun, Yun, Su Yeoung, Han, Ji Young, Yoo, Jong Shin, An, Hyun Joo, Park, Young Mok
Format: Article
Language:English
Subjects:
14-3-3 Proteins - analysis
14-3-3 Proteins - metabolism
Analytical Chemistry
Biochemistry
Bone marrow
Cell culture
Cell Differentiation
Cells, Cultured
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
Chemistry, Analytic
Differentiation
Electrophoresis, Polyacrylamide Gel
Fibroblast Growth Factor 2 - pharmacology
Fibroblasts
Food Science
Growth factors
Human
Humans
Kinases
Laboratory Medicine
Mass spectrometry
Mass Spectrometry - methods
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - drug effects
Mesenchymal Stromal Cells - metabolism
Monitoring/Environmental Analysis
Multipotent Stem Cells - metabolism
Musculoskeletal system
Original Paper
Physiological aspects
Proteins
Proteins - analysis
Proteins - classification
Proteins - metabolism
Proteome - analysis
Proteomics - methods
Quantitative
Receptors, Laminin - metabolism
Reproducibility of Results
Scientific imaging
Searching
Signal transduction
Sodium
Spectrometry, Mass, Electrospray Ionization
Stem cells
Tandem Mass Spectrometry
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Summary: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. Figure ᅟ
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-013-6969-z