Forschungs- & Entwicklungsinformationsdienst der Gemeinschaft - CORDIS

Generation of encapsulated cells expressing GDNF

To generate cell lines secreting GDNF suitable for encapsulation, stable clones were established from ARPE-19 cells (immortalized retinal pigment epithelial cells) by transfection with two different GDNF expression constructs. More than 200 clones were screened and among those, six clones secreting 90-133 ngGDNF/105 cells/24h in serum supplemented growth medium were chosen for further evaluation. When culturing the clones in a serum-free medium optimized for epithelial cells, GDNF secretion was increased approximately 2 folds for most clones. For all clones, stable GDNF secretion appeared to be maintained over 4 months in culture (and more than 20 passages) under proliferative conditions. Furthermore, GDNF secretion was stable at least 6 weeks after reaching confluency and GDNF bioactivity of conditioned media from the clones has been verified using a cell-based assay. After encapsulation of the clones in a standard device for experimental implantation into rat brains, GDNF release levels between 1-15 ng/device/24h could be achieved. Such devices could be used to obtain mechanistic insights into the neuroprotective role in appropriate animal models.

Verwandte Informationen

Reported by

Pederstrupvej 93
See on map
Folgen Sie uns auf: RSS Facebook Twitter YouTube Verwaltet vom Amt für Veröffentlichungen der EU Nach oben