Differential expression of cell surface proteins in human bone marrow mesenchymal stem cells cultured with or without basic fibroblast growth factor containing medium.
Mesenchymal stem cells (MSCs) are multipotent cells, which have the capability to differentiate into various mesenchymal tissues such as bone, cartilage, fat, tendon, muscle, and marrow stroma. However, they lose the capability of multi-lineage differentiation after several passages. It is known that basic fibroblast growth factor (bFGF) increases growth rate, differentiation potential, and morphological changes of MSCs in vitro. In this report, we have used 2-DE coupled to MS to identify differentially expressed proteins at the cell membrane level in MSCs growing in bFGF containing medium. The cell surface proteins isolated by the biotin-avidin affinity column were separated by 2-DE in triplicate experiments. A total of 15 differentially expressed proteins were identified by quadrupole-time of flight tandem MS. Nine of the proteins were upregulated and six proteins were downregulated in the MSCs cultured with bFGF containing medium. The expression level of three actin-related proteins, F-actin-capping protein subunit alpha-1, actin-related protein 2/3 complex subunit 2, and myosin regulatory light chain 2, was confirmed by Western blot analysis. The results indicate that the expression levels of F-actin-capping protein subunit alpha-1, actin-related protein 2/3 complex subunit 2, and myosin regulatory light chain 2 are important in bFGF-induced morphological change of MSCs.
Lee
Sang Kwang
SK
Mass Spectrometry Research Center, Korea Basic Science Institute, Daejeon, Republic of Korea.
Kim
Yongtae
Y
Kim
Sung-Soo
SS
Lee
Jeong Hwa
JH
Cho
Kun
K
Lee
Sang Sook
SS
Lee
Zee-Won
ZW
Kwon
Kyung-Hoon
KH
Kim
Young Hye
YH
Suh-Kim
Haeyoung
H
Yoo
Jong Shin
JS
Park
Young Mok
YM
D000199:Actins / Q000378:metabolism; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media; D015180:Electrophoresis, Gel, Two-Dimensional; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000096:biosynthesis*; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
D000199:Actins; D003470:Culture Media; D008565:Membrane Proteins; D011506:Proteins; D016222:Fibroblast Growth Factor 2