Purified mouse dopamine neurons thrive and function after transplantation into brain but require novel glial factors for survival in culture.
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.
Donaldson
A E
AE
Farber Institute for Neurosciences, Thomas Jefferson University Medical College, Philadelphia, PA 19107, USA.
Marshall
C E
CE
Yang
Ming
M
Suon
S
S
Iacovitti
Lorraine
L
D000818:Animals; D016380:Brain Tissue Transplantation; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004298:Dopamine / Q000502:physiology; D016332:Fetal Tissue Transplantation; D005434:Flow Cytometry; D006085:Graft Survival; D049452:Green Fluorescent Proteins / Q000235:genetics; D006133:Growth Substances / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000502:physiology / Q000637:transplantation*; D010300:Parkinson Disease / Q000628:therapy*; D051381:Rats; D011916:Rats, Inbred F344
D017077:Culture Media, Conditioned; D006133:Growth Substances; D049452:Green Fluorescent Proteins; D004298:Dopamine