Original Paper

Analytical and Bioanalytical Chemistry

, Volume 405, Issue 16, pp 5501-5517

First online:

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

  • Sang Kwang LeeAffiliated withMass Spectrometry Research Center, Korea Basic Science InstituteEulji Medical and Biological Research Institute (EMBRI)
  • , Jae Ho KimAffiliated withMass Spectrometry Research Center, Korea Basic Science Institute
  • , Sung-Soo KimAffiliated withDepartment of Anatomy, Ajou University, School of Medicine
  • , Taewook KangAffiliated withMass Spectrometry Research Center, Korea Basic Science InstituteGraduate Schools of Analytical Science and Technology, Chungnam National University
  • , Nam Hyun ParkAffiliated withMass Spectrometry Research Center, Korea Basic Science InstituteGraduate Schools of Analytical Science and Technology, Chungnam National University
  • , Kyung-Hoon KwonAffiliated withMass Spectrometry Research Center, Korea Basic Science Institute
  • , Sang Sook LeeAffiliated withBiotechnology Fusion Research Team, Korea Basic Science InstituteDepartment of Biology, Chungnam National University
  • , Zee Won LeeAffiliated withBiotechnology Fusion Research Team, Korea Basic Science Institute
  • , Hae young Suh-KimAffiliated withDepartment of Anatomy, Ajou University, School of Medicine
    • , Kun ChoAffiliated withMass Spectrometry Research Center, Korea Basic Science Institute
    • , Su Yeoung YunAffiliated withMass Spectrometry Research Center, Korea Basic Science InstituteGraduate Schools of Analytical Science and Technology, Chungnam National University
    • , Ji Young HanAffiliated withMass Spectrometry Research Center, Korea Basic Science Institute
    • , Jong Shin YooAffiliated withMass Spectrometry Research Center, Korea Basic Science InstituteGraduate Schools of Analytical Science and Technology, Chungnam National University
    • , Hyun Joo AnAffiliated withGraduate Schools of Analytical Science and Technology, Chungnam National University
    • , Young Mok ParkAffiliated withMass Spectrometry Research Center, Korea Basic Science InstituteBiotechnology Fusion Research Team, Korea Basic Science InstituteMass Spectrometry Research Center, Korea Basic Science Institute Email author 

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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.

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Keywords

Basic fibroblast growth factor Mesenchymal stem cell Proteome Surface protein