Nonviral glial cell-derived neurotrophic factor gene transfer enhances survival of cultured dopaminergic neurons and improves their function after transplantation in a rat model of Parkinson's disease.
Transplantation of dopaminergic fetal mesencephalic tissue into the striatum is currently being developed for treatment of patients with advanced Parkinson's disease. Ethical concerns regarding the use of human fetal tissue, and the limited availability as well as poor survival and differentiation of dopaminergic neurons after transplantation have reduced the extent and outcome of this approach so far. With the purpose of finding means to increase the yield of dopaminergic neurons in transplants, and to reduce the amount of fetal tissue needed for each transplanted patient, we transfected rat fetal ventral mesencephalic (VM) tissue grown as organotypic free-floating roller tube (FFRT) cultures with a vector encoding human glial cell-derived neurotrophic factor (hGDNF). For transfer of an episomal expression vector (pRep7-GDNF8) a nonviral, nonliposomal cationic transfection technique was applied and optimized. Recombinant hGDNF expression resulted in a higher number of TH-positive neurons in the cultures as measured 6 days after transfection. Ventral mesencephalic cultures expressing hGDNF were then grafted into the striatum of unilaterally 6-hydroxydopamine (6-OHDA)-lesioned rats. Grafting of genetically modified VM cultures resulted in earlier functional recovery compared with grafting nontransfected cultures. We conclude that organotypic free-floating roller tube cultures can be successfully transfected to produce hGDNF with effects on TH-expressing neurons in vitro and functional effects after grafting in a rat Parkinson's disease model.
Bauer
M
M
Department of Neurology, Klinikum Grosshadern, Ludwig Maximilians Universität München, Munich, Germany.
Meyer
M
M
Grimm
L
L
Meitinger
T
T
Zimmer
J
J
Gasser
T
T
Ueffing
M
M
Widmer
H R
HR
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006801:Humans; D008636:Mesencephalon; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009457:Neuroglia; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D009924:Organ Culture Techniques; D016627:Oxidopamine / Q000378:metabolism*; D010300:Parkinson Disease; D051381:Rats; D017207:Rats, Sprague-Dawley; D016378:Tissue Transplantation / Q000379:methods; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
C495039:GDNF protein, human; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D016627:Oxidopamine; D001616:beta-Galactosidase