title: Purified mouse dopamine neurons thrive and function after transplantation into brain but require novel glial factors for survival in culture.
abstract: Cell replacement therapy in Parkinson's disease depends on a reliable source of purified dopamine (DA) neurons (PDN) and the identification of factors relevant to their survival. Our goal was to genetically tag and purify by flow cytometry embryonic midbrain DA neurons from a transgenic mouse line carrying 11 kb of human tyrosine hydroxylase promoter driving expression of the enhanced green fluorescent protein(GFP) for studies in vivo and in vitro. A 99% purification of GFP+ cells was achieved. When transplanted into 6-hydroxydopamine-treated rat striatum, PDN survived, became well-integrated and produced recovery from amphetamine-induced motor behaviors. However, when grown in culture, PDN died within days of plating. No known growth factors prevented PDN death as did incubation with novel factors in glia/glial-conditioned media. We conclude that GFP-tagged DA neurons can be purified to homogeneity and can survive and function when grown with glial factors in vitro or after transplantation in vivo.
authors.affiliation: Farber Institute for Neurosciences, Thomas Jefferson University Medical College, Philadelphia, PA 19107, USA.
authors.forename: A E
authors.initials: AE
authors.lastname: Donaldson
authors.forename: C E
authors.initials: CE
authors.lastname: Marshall
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.forename: S
authors.initials: S
authors.lastname: Suon
authors.forename: Lorraine
authors.initials: L
authors.lastname: Iacovitti
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D016380:Brain Tissue Transplantation / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004298:Dopamine / Q000378:metabolism*; D005434:Flow Cytometry; D006085:Graft Survival / Q000187:drug effects / Q000502:physiology*; D049452:Green Fluorescent Proteins; D006801:Humans; D008636:Mesencephalon / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D051379:Mice; D008822:Mice, Transgenic; D009414:Nerve Growth Factors / Q000378:metabolism*; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D016627:Oxidopamine; D020734:Parkinsonian Disorders / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
chemical_list: D009414:Nerve Growth Factors; D049452:Green Fluorescent Proteins; D016627:Oxidopamine; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine