![[feed]](/style/images/feed-icon-14x14.png){kind=icon}
Tools
Tools
| Item Type: | Article |
|---|---|
| Title: | Lipid-mediated gene transfer of acidic fibroblast growth factor into human corneal endothelial cells |
| Creators Name: | Dannowski, H., Bednarz, J., Reszka, R., Engelmann, K. and Pleyer, U. |
| Abstract: | The aim of this study was to optimize non-viral gene transfer conditions and investigate the effect of fibroblast growth factor-1 (FGF-1) gene transfer on human corneal endothelial cell (HCEC) proliferation. Five non-viral vectors (Lipofectin™, DMRIE-C™, DAC-30, Effectene™, FuGene™6) were used to transfect HCEC with plasmids coding for enhanced green fluorescent protein (EGFP) and FGF-1. Transfection efficiency and toxicity (n=6) were quantified and optimized using the EGFP construct by FACS-analysis. Using optimal conditions HCEC were transfected with the FGF-1 plasmid and cell proliferation as well as expression of FGF-1 were determined at days 4 and 7 by counting and western blotting, respectively. Lipofectin (17±2·02%) transfected HCEC more successfully than DMRIE-C (11±1·46%), Effectene (9±0·62%), FuGene (9±0·93%) and DAC-30 (7±0·59%). Toxicity of the lipids ranged from 2 to 4%. Optimal HCEC proliferation was achieved with DAC-30/FGF-1 (P<0·05), whereas all other vectors did not result in significantly increased cell proliferation. However, all of the transfected cells produced FGF-1 in different amounts as indicated by western blotting. Efficient and almost non-toxic transfer of the FGF-1 gene into HCEC can be successfully achieved by lipid-based techniques. Using optimal conditions significantly increased cell proliferation was independent on gene transfer efficiency. This may indicate that even a low transfection rate is sufficient to produce a concentration of FGF-1 that will have a stimulatory effect on HCECs. |
| Keywords: | Cell Proliferation, Corneal Endothelial Cells, Fibroblast Growth Factor, Gene Transfer, Liposomes |
| Source: | Experimental Eye Research |
| ISSN: | 0014-4835 |
| Publisher: | Ademic Press Ltd Elsevier Science Ltd |
| Volume: | 80 |
| Number: | 1 |
| Page Range: | 93-101 |
| Date: | 1 January 2005 |
| Official Publication: | https://doi.org/10.1016/j.exer.2004.08.024 |
| PubMed: | View item in PubMed |
Repository Staff Only: item control page
