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Journal of Biochemistry Advance Access originally published online on October 23, 2007
Journal of Biochemistry 2007 142(6):741-748; doi:10.1093/jb/mvm189
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© 2007 The Japanese Biochemical Society.

Effect of Forced Expression of Basic Fibroblast Growth Factor in Human Bone Marrow-Derived Mesenchymal Stromal Cells

Masahiro J. Go1,2,*, Chiemi Takenaka1,2 and Hajime Ohgushi1

1National Institute of Advanced Industrial Science and Technology, Research Institute for Cell Engineering, Tissue Engineering Research Group, Nakouji 3-11-46, Amagasaki, Hyogo 661-0974; and 2Stem Cell Sciences KK, Minatojima-Minami 5-5-2, Kobe, Hyogo 650-0047, Japan

*To whom correspondence should be addressed. Tel: + 81-6-6494-7807, Fax: +81-6-6494-7861, E-mail: mj-gou{at}aist.go.jp or mjgo{at}bca.bai.ne.jp

Received August 19, 2007; Accepted September 29, 2007


  Abstract

Mesenchymal stromal cells (MSCs) derived from human bone marrow are expected to be utilized for the purpose of tissue engineering, because of their extensive self-renewal or proliferation capability. The capability decreases after several passages, however. Basic fibroblast growth factor (bFGF) is commonly used for culture of various cells including bone marrow-derived MSCs. With the aim of conferring higher capability on human bone marrow MSCs, we introduced the bFGF gene into the passaged cells by retroviral system. The bFGF-expressing MSCs, even at 7 to 8 passages after the infection, showed consistent proliferation capability. The capability was not detected in control cells even in culture media containing the bFGF protein. Thus, we could not mimic the effect of forced expression of bFGF by exogenously adding the bFGF protein in culture media. Although we expressed the shortest isoform of bFGF, which was considered to be mostly cytosolic, we found the protein mostly in the nucleus. Our observations demonstrate not only an effective way to maintain proliferation potentials of MSCs, but also a possibility that there may be mechanistic and functional differences in the signal transduction events between endogenously expressed and exogenously added bFGF protein in MSCs.

Key Words: basic FGF, bone marrow, forced expression, mesenchymal cells, retrovirus

Abbreviations: bFGF, basic fibroblast growth factor; EGFP, enhanced green fluorescent protein; FBS, fetal bovine serum; IRES, internal ribosome entry site; MSC, mesenchymal stromal cell; PBS, phosphate-buffered saline


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