PuSH - Publication Server of Helmholtz Zentrum München: Lipid-Mediated Glial Cell Line-Derived Neurotrophic Factor Gene Transfer to Cultured Porcine Ventral Mesencephalic Tissue.

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Bauer, M. ; Meyer, M.* ; Brevig, Th.* ; Gasser, Th.* ; Widmer, H.R.* ; Zimmer, J.* ; Ueffing, M.

Lipid-Mediated Glial Cell Line-Derived Neurotrophic Factor Gene Transfer to Cultured Porcine Ventral Mesencephalic Tissue.

Exp. Neurol. 177, 40-49 (2002)

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Open Access Green as soon as Postprint is submitted to ZB.

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Transplantation of dopaminergic ventral mesencephalic (VM) tissue into the basal ganglia of patients with Parkinson's disease (PD) shows at best moderate symptomatic relief in some of the treated cases. Experimental animal studies and clinical trials with allogenic and xenogenic pig-derived VM tissue grafts to PD patients indicate that one reason for the poor outcome of neural transplantation is the low survival and differentiation of grafted dopaminergic neurons. To improve dopaminergic cell survival through a genetherapeutic approach we have established and report here results of lipid-mediated transfer of the gene for human glial cell line-derived neurotrophic factor (GDNF) to embryonic (E27/28) porcine VM tissue kept as organotypic explant cultures. Treatment of the developing VM with two mitogens, basic fibroblast growth factor and epidermal growth factor, prior to transfection significantly increased transfection yields. Expression of human GDNF via an episomal vector could be detected by in situ hybridization and by the measuring of GDNF protein secreted into the culture medium. When compared to mock-transfected controls, VM tissue expressing recombinant GDNF contained significantly higher numbers of tyrosine hydroxylase-positive neurons in the cultured VM tissue. We conclude that lipid-mediated gene transfer employed on embryonic pig VM explant cultures is a safe and effective method to improve survival of dopamitnergic neurons and may become a valuable tool to improve allo- and xenotransplantation treatment in Parkinson's disease.

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