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dc.contributor.authorNiclis, JC
dc.contributor.authorGantner, CW
dc.contributor.authorAlsanie, WF
dc.contributor.authorMcDougall, SJ
dc.contributor.authorBye, CR
dc.contributor.authorElefanty, AG
dc.contributor.authorStanley, EG
dc.contributor.authorHaynes, JM
dc.contributor.authorPouton, CW
dc.contributor.authorThompson, LH
dc.contributor.authorParish, CL
dc.date.accessioned2020-12-21T01:52:21Z
dc.date.available2020-12-21T01:52:21Z
dc.date.issued2017-03-01
dc.identifierpii: sctm.2016-0073
dc.identifier.citationNiclis, J. C., Gantner, C. W., Alsanie, W. F., McDougall, S. J., Bye, C. R., Elefanty, A. G., Stanley, E. G., Haynes, J. M., Pouton, C. W., Thompson, L. H. & Parish, C. L. (2017). Efficiently Specified Ventral Midbrain Dopamine Neurons from Human Pluripotent Stem Cells Under Xeno-Free Conditions Restore Motor Deficits in Parkinsonian Rodents. STEM CELLS TRANSLATIONAL MEDICINE, 6 (3), pp.937-948. https://doi.org/10.5966/sctm.2016-0073.
dc.identifier.issn2157-6564
dc.identifier.urihttp://hdl.handle.net/11343/256694
dc.description.abstract: Recent studies have shown evidence for the functional integration of human pluripotent stem cell (hPSC)-derived ventral midbrain dopamine (vmDA) neurons in animal models of Parkinson's disease. Although these cells present a sustainable alternative to fetal mesencephalic grafts, a number of hurdles require attention prior to clinical translation. These include the persistent use of xenogeneic reagents and challenges associated with scalability and storage of differentiated cells. In this study, we describe the first fully defined feeder- and xenogeneic-free protocol for the generation of vmDA neurons from hPSCs and utilize two novel reporter knock-in lines (LMX1A-eGFP and PITX3-eGFP) for in-depth in vitro and in vivo tracking. Across multiple embryonic and induced hPSC lines, this "next generation" protocol consistently increases both the yield and proportion of vmDA neural progenitors (OTX2/FOXA2/LMX1A) and neurons (FOXA2/TH/PITX3) that display classical vmDA metabolic and electrophysiological properties. We identify the mechanism underlying these improvements and demonstrate clinical applicability with the first report of scalability and cryopreservation of bona fide vmDA progenitors at a time amenable to transplantation. Finally, transplantation of xeno-free vmDA progenitors from LMX1A- and PITX3-eGFP reporter lines into Parkinsonian rodents demonstrates improved engraftment outcomes and restoration of motor deficits. These findings provide important and necessary advancements for the translation of hPSC-derived neurons into the clinic. SIGNIFICANCE: The authors report the generation of highly pure midbrain dopamine cultures under feeder-free, fully defined, and xeno-free conditions from human pluripotent stem cells. Xeno-free differentiated cells display gene, protein, and electrophysiological properties of midbrain neurons, as well as improved grafting outcomes in Parkinsonian rodents, observations enhanced by the use of two novel reporter lines of interest to this research field. Furthermore, for the first time, ventral midbrain dopamine neurons were amenable to scalability and cryopreservation, crucial steps for the advancement of cell replacement therapy in Parkinson's disease.
dc.languageEnglish
dc.publisherWILEY
dc.titleEfficiently Specified Ventral Midbrain Dopamine Neurons from Human Pluripotent Stem Cells Under Xeno-Free Conditions Restore Motor Deficits in Parkinsonian Rodents
dc.typeJournal Article
dc.identifier.doi10.5966/sctm.2016-0073
melbourne.affiliation.departmentFlorey Department of Neuroscience and Mental Health
melbourne.affiliation.departmentAnatomy and Neuroscience
melbourne.affiliation.departmentPaediatrics (RCH)
melbourne.source.titleStem Cells Translational Medicine
melbourne.source.volume6
melbourne.source.issue3
melbourne.source.pages937-948
dc.rights.licenseCC BY
melbourne.elementsid1103947
melbourne.contributor.authorThompson, Lachlan
melbourne.contributor.authorParish, Clare
melbourne.contributor.authorMcDougall, Stuart
melbourne.contributor.authorElefanty, Andrew
melbourne.contributor.authorAlsanie, Walaa
melbourne.contributor.authorGantner, Carlos William Baevski
melbourne.contributor.authorNICLIS, JONATHAN
melbourne.contributor.authorBye, Chris
melbourne.contributor.authorStanley, Edouard
dc.identifier.eissn2157-6580
melbourne.accessrightsOpen Access


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