Ex vivo infection of human embryonic spinal cord neurons prior to transplantation into adult mouse cord
1 Department of Ophthalmology, Faculty of General Medicine, University of Szeged, Szeged, Hungary
2 Department of Medical Biology, Faculty of General Medicine, University of Szeged, Szeged, Hungary
3 Laboratory of Molecular Neuroendocrinology, Insitute of Experimental Medicine, Budapest, Hungary
4 Department of Virology, Central Veterinary Institute, Budapest, Hungary
5 Dept. of Pharmacology, University of Oxford, Mansfield Road, Oxford, OX1 3QT, UK
BMC Neuroscience 2010, 11:65 doi:10.1186/1471-2202-11-65Published: 29 May 2010
Genetically modified pseudorabies virus (Prv) proved suitable for the delivery of foreign genes to rodent embryonic neurons ex vivo and maintaining foreign gene expression after transplantation into spinal cord in our earlier study. The question arose of whether human embryonic neurons, which are known to be more resistant to Prv, could also be infected with a mutant Prv. Specifically, we investigated whether a mutant Prv with deleted ribonucleotide reductase and early protein 0 genes has the potential to deliver marker genes (gfp and β-gal) into human embryonic spinal cord neurons and whether the infected neurons maintain expression after transplantation into adult mouse cord.
The results revealed that the mutant Prv effectively infected human embryonic spinal cord neurons ex vivo and the grafted cells exhibited reporter gene expression for several weeks. Grafting of infected human embryonic cells into the spinal cord of immunodeficient (rnu-/rnu-) mice resulted in the infection of some of the host neurons.
These results suggest that Prv is suitable for the delivery of foreign genes into transplantable human cells. This delivery method may offer a new approach to use genetically modified cells for grafting in animal models where spinal cord neuronal loss or axon degeneration occurs.