Localized and sustained SDF-1 gene release mediated by fibronectin films: A potential method for recruiting stem cells.
Gene-activated matrix has wide potential utilization in tissue engineering. It may genetically modify cells with plasmid DNA encoding therapeutic genes and allow sustained expression and release of the proteins to surrounding tissues. In this study, we assessed the feasibility of the local gene release from human fibronectin (HFN) substrate and the efficacy of local release of stromal cell-derived factor-1 (SDF-1) gene on c-kit+ cell homing. Cationic polymer polyethylenimine (25kDa PEI) was used as non-viral DNA vector. Gene-activated HFN (GAH) was prepared by mixing PEI/DNA complexes with HFN substrate. The DNA retardation, the complex size, and the DNA release speed from the GAH were studied. The in vitro transfection was optimized by luciferase expression and cell viability assay in the COS7 cell line. Localized gene expression in COS7 cells cultured on the GAH was assessed by LacZ and GFP-N3-SDF-1 marker genes. Ckit+ cell homing was investigated in response to the local in vitro SDF-1 expression from rat mesenchymal stem cells (RMSCs) cultured on GAH. Results showed GAH allows long time-sustained DNA release, localized gene delivery, and high transfection efficiency. Local SDF-1 expression with GAH is a promising method to induce targetable stem cell homing.
Department of Cardiac Surgery, University of Rostock, Rostock - Germany.
Weiwei
W
Wang
Wenzhong
W
Li
Lee-L
LL
Ong
Karola
K
Lutzow
Andreas
A
Lendlein
Dario
D
Furlani
Ralf
R
Gabel
Deling
D
Kong
Jun
J
Wang
Ren-K
RK
Li
Gustav
G
Steinhoff
Nan
N
Ma
D000818:Animals; D019556:COS Cells; D002465:Cell Movement; D002470:Cell Survival; D054377:Chemokine CXCL12 / Q000096:biosynthesis* / Q000235:genetics; D002522:Chlorocebus aethiops; D024202:Electrophoretic Mobility Shift Assay; D005240:Feasibility Studies; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D007097:Imines / Q000737:chemistry; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D053758:Nanoparticles; D011095:Polyethylenes / Q000737:chemistry; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*
D054377:Chemokine CXCL12; D005353:Fibronectins; D007097:Imines; D011095:Polyethylenes; D011993:Recombinant Fusion Proteins; C505405:poly(ethylene imine); D019009:Proto-Oncogene Proteins c-kit