title: Polymer-encapsulated PC-12 cells demonstrate high-affinity uptake of dopamine in vitro and 18F-Dopa uptake and metabolism after intracerebral implantation in nonhuman primates.
abstract: Intracranial implantation of polymer-encapsulated PC-12 cells has been shown to improve motor behavioral performance in animal models of Parkinson's disease. The purpose of this blinded study was to examine whether such improvement is associated with the active uptake and metabolism of dopamine precursors by intracerebrally implanted polymer-encapsulated PC-12 cells. In an in vitro experiment we demonstrate that 3H-dopamine uptake by PC-12 cells was 10(8) fmol/min x 10(6) cells, and that this uptake can be specifically blocked 88% by the addition of 10nM of nomifensine. In the in vivo experiments, polymer-encapsulated PC-12 cells were implanted in four MPTP-treated monkeys into the left deep parietal white matter (R1) or left striatum (R2-4). A fifth MPTP-treated monkey (R5) served as a control and received left striatal implants of empty capsules. 18-F-Dopa Positron Emission Tomography (PET) imaging was performed on each monkey before and after implantation surgery by blinded investigators. PET images obtained 5-13 wk after implantation demonstrated well delineated focal areas of high 18F-dopa uptake in R1, R2, and R4. The focal area of high 18F-dopa uptake in R1 precisely coregistered on a brain magnetic resonance image to the site of implantation. R3 (in whom the polymer-encapsulated PC-12 cells demonstrated poor cell survival upon explantation) and R5 (empty capsules) failed to demonstrate any area of increased 18F-dopa uptake in their PET images. Histological examination of the host brain revealed no sprouting of dopaminergic nerve terminals around the implantation sites of the polymer-encapsulated PC-12 cells. These results indicate that the previously noted behavioral improvement after intrastriatal implantation of polymer encapsulated PC-12 cells is at least in part due to their highly specific uptake and metabolism of dopamine precursors. Furthermore, these data suggest that polymer-encapsulated PC-12 cells can store, reuptake, and functionally replenish dopamine and therefore, may be an effective treatment for Parkinson's disease.
authors.affiliation: Department of Neurology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: T
authors.initials: T
authors.lastname: Subramanian
authors.forename: D F
authors.initials: DF
authors.lastname: Emerich
authors.forename: R A
authors.initials: RA
authors.lastname: Bakay
authors.forename: J M
authors.initials: JM
authors.lastname: Hoffman
authors.forename: M M
authors.initials: MM
authors.lastname: Goodman
authors.forename: T M
authors.initials: TM
authors.lastname: Shoup
authors.forename: G W
authors.initials: GW
authors.lastname: Miller
authors.forename: A I
authors.initials: AI
authors.lastname: Levey
authors.forename: G W
authors.initials: GW
authors.lastname: Hubert
authors.forename: S
authors.initials: S
authors.lastname: Batchelor
authors.forename: S R
authors.initials: SR
authors.lastname: Winn
authors.forename: J A
authors.initials: JA
authors.lastname: Saydoff
authors.forename: R L
authors.initials: RL
authors.lastname: Watts
mesh_terms: D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000000981:diagnostic imaging / Q000378:metabolism; D002214:Capsules; D002352:Carrier Proteins / Q000032:analysis; D017690:Cell Transplantation / Q000379:methods*; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors / Q000494:pharmacology; D005462:Fluorine Radioisotopes; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D008253:Macaca mulatta; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D009627:Nomifensine / Q000494:pharmacology; D016716:PC12 Cells / Q000378:metabolism* / Q000637:transplantation*; D010302:Parkinson Disease, Secondary / Q000139:chemically induced / Q000000981:diagnostic imaging; D011108:Polymers; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D014055:Tomography, Emission-Computed; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis
chemical_list: D002214:Capsules; D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors; D005462:Fluorine Radioisotopes; D005904:Glial Fibrillary Acidic Protein; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D011108:Polymers; D009627:Nomifensine; C043437:fluorodopa F 18; D004295:Dihydroxyphenylalanine; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D014446:Tyrosine 3-Monooxygenase; D011188:Potassium; D004298:Dopamine