Analytical and Bioanalytical Chemistry

, Volume 405, Issue 16, pp 5501–5517

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

  • Sang Kwang Lee
  • Jae Ho Kim
  • Sung-Soo Kim
  • Taewook Kang
  • Nam Hyun Park
  • Kyung-Hoon Kwon
  • Sang Sook Lee
  • Zee Won Lee
  • Hae young Suh-Kim
  • Kun Cho
  • Su Yeoung Yun
  • Ji Young Han
  • Jong Shin Yoo
  • Hyun Joo An
  • Young Mok Park
Original Paper

DOI: 10.1007/s00216-013-6969-z

Cite this article as:
Lee, S.K., Kim, J.H., Kim, SS. et al. Anal Bioanal Chem (2013) 405: 5501. doi:10.1007/s00216-013-6969-z

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.

https://static-content.springer.com/image/art%3A10.1007%2Fs00216-013-6969-z/MediaObjects/216_2013_6969_Figa_HTML.gif
Figure

Keywords

Basic fibroblast growth factorMesenchymal stem cellProteomeSurface protein

Supplementary material

216_2013_6969_MOESM1_ESM.pdf (725 kb)
ESM 1(PDF 724 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Sang Kwang Lee
    • 1
    • 6
  • Jae Ho Kim
    • 1
  • Sung-Soo Kim
    • 2
  • Taewook Kang
    • 1
    • 3
  • Nam Hyun Park
    • 1
    • 3
  • Kyung-Hoon Kwon
    • 1
  • Sang Sook Lee
    • 4
    • 7
  • Zee Won Lee
    • 4
  • Hae young Suh-Kim
    • 2
  • Kun Cho
    • 1
  • Su Yeoung Yun
    • 1
    • 3
  • Ji Young Han
    • 1
  • Jong Shin Yoo
    • 1
    • 3
  • Hyun Joo An
    • 3
  • Young Mok Park
    • 1
    • 4
    • 5
  1. 1.Mass Spectrometry Research CenterKorea Basic Science InstituteOchangRepublic of Korea
  2. 2.Department of Anatomy, Ajou UniversitySchool of MedicineSuwonRepublic of Korea
  3. 3.Graduate Schools of Analytical Science and TechnologyChungnam National UniversityDaejeonRepublic of Korea
  4. 4.Biotechnology Fusion Research TeamKorea Basic Science InstituteDaejeonRepublic of Korea
  5. 5.Mass Spectrometry Research CenterKorea Basic Science InstituteOchangRepublic of Korea
  6. 6.Eulji Medical and Biological Research Institute (EMBRI)DaejeonRepublic of Korea
  7. 7.Department of BiologyChungnam National UniversityDaejeonRepublic of Korea