| Author/Editor | Čemažar, Maja; Wilson, Ian; Dachs, Gabi U; Tozer, Gillian M; Serša, Gregor | |
| Title | Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy - spatial and time dependent distribution | |
| Type | članek | |
| Source | BMC Cancer | |
| Vol. and No. | Letnik 4, št. 81 | |
| Publication year | 2004 | |
| Volume | str. 1-7 | |
| Language | eng | |
| Abstract | Background: Electroporation is currendy receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efflciency of electroporation-assisced DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost imporiance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. Methods: Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP) and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisced DNA delivery using two different sets of eleccric pulse parameters were compared. Results: Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, I Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the mmors up to 6 days posi-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was deiected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. (Abstract truncated at 2000 characters). | |
| Descriptors | CARCINOSARCOMA DNA ELECTROPORATION TRANSFECTION GENE THERAPY FLUORESCENT DYES LIPOSOMES PLASMIDS MICROSCOPY, FLUORESCENCE |