Nonviral delivery of basic fibroblast growth factor gene to bone marrow stromal cells.
Basic fibroblast growth factor (bFGF) is capable of stimulating osteogenic differentiation of preosteoblast cells in vitro and new bone tissue deposition in vivo. Delivering the gene for the protein, rather than the protein itself, is considered advantageous for bone repair since gene delivery obviates the need to produce the protein in pharmaceutical quantities. To explore the feasibility of bFGF gene delivery by nonviral methods, we transfected primary rat bone marrow stromal cells (BMSC) using cationic polymers (polyethylenimine and poly(L-lysine)-palmitic acid) in vitro. After delivering a bFGF-expression plasmid (pFGF2-IRES-AcGFP) to BMSC, the presence of bFGF in culture supernatants was detected by a commercial ELISA. As much as 0.3 ng bFGF/10(6) cells/day was obtained from the BMSC under optimal conditions. This secretion rate was approximately 100-fold lower than the secretion obtained from immortal, and easy-to-transfect, human 293T cells. These data suggest the feasibility of modifying BMSC with nonviral delivery systems for bFGF expression, but also highlight the need for substantial improvement in transfection rate for an effective therapy.
Clements
Başak Açan
BA
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Hsu
Charlie Y M
CY
Kucharski
Cezary
C
Lin
Xiaoyue
X
Rose
Laura
L
Uludağ
Hasan
H
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D010012:Osteogenesis; D011094:Polyethyleneimine / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D051381:Rats; D017154:Stromal Cells / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection / Q000379:methods*
D003470:Culture Media; D016222:Fibroblast Growth Factor 2; D011107:Polylysine; D011094:Polyethyleneimine; D000469:Alkaline Phosphatase