Glial cell line-derived neurotrophic factor (
GDNF) is one of the most important
proteins playing a pivotal role in growing and repairing of the nervous system.
GDNF therapy is one of the suggested options in the treatment of
neurodegenerative diseases. Limitations in the viral gene delivery and its side effects after
therapy have encouraged us to use a non-viral method one for this purpose. We transfected rat bone marrow stromal cells (BMSCs) in ex vivo conditions using
Lipofectamine 2000 reagent with pEGFP-C1 and a constructed vector carrying the human proGDNF (pSecTag2/HygroB-human proGDNF), transiently and stably, respectively. The rate of transient transfection of rat BMSCs was eight percent and transfected rat BMSCs with pSecTag2/HygroB-human proGDNF stabilized by adding
Hygromycin B in cell culture medium at 200 μg/ml. Semi-quantitative data analysis from Western-blot technique showed that stable transfected cells secrete
GDNF at higher level in comparison with control cells (6.530 fold in the supernatant). The present study supports the utility of
liposome-mediated transfection for overexpressing human
GDNF in rat BMSCs. For this purpose and in order to get more yield of human
GDNF secretion from the stable transfected rat BMSCs, we used a vector containing another
signal sequence instead of its own pre-segment of proGDNF
protein. This is the first report in this regard and the data presented will be potentially useful for human gene transfer
therapies in a variety of
neurodegenerative diseases.