title: Improved cell therapy protocols for Parkinson's disease based on differentiation efficiency and safety of hESC-, hiPSC-, and non-human primate iPSC-derived dopaminergic neurons.
abstract: The main motor symptoms of Parkinson's disease are due to the loss of dopaminergic (DA) neurons in the ventral midbrain (VM). For the future treatment of Parkinson's disease with cell transplantation it is important to develop efficient differentiation methods for production of human iPSCs and hESCs-derived midbrain-type DA neurons. Here we describe an efficient differentiation and sorting strategy for DA neurons from both human ES/iPS cells and non-human primate iPSCs. The use of non-human primate iPSCs for neuronal differentiation and autologous transplantation is important for preclinical evaluation of safety and efficacy of stem cell-derived DA neurons. The aim of this study was to improve the safety of human- and non-human primate iPSC (PiPSC)-derived DA neurons. According to our results, NCAM(+) /CD29(low) sorting enriched VM DA neurons from pluripotent stem cell-derived neural cell populations. NCAM(+) /CD29(low) DA neurons were positive for FOXA2/TH and EN1/TH and this cell population had increased expression levels of FOXA2, LMX1A, TH, GIRK2, PITX3, EN1, NURR1 mRNA compared to unsorted neural cell populations. PiPSC-derived NCAM(+) /CD29(low) DA neurons were able to restore motor function of 6-hydroxydopamine (6-OHDA) lesioned rats 16 weeks after transplantation. The transplanted sorted cells also integrated in the rodent brain tissue, with robust TH+/hNCAM+ neuritic innervation of the host striatum. One year after autologous transplantation, the primate iPSC-derived neural cells survived in the striatum of one primate without any immunosuppression. These neural cell grafts contained FOXA2/TH-positive neurons in the graft site. This is an important proof of concept for the feasibility and safety of iPSC-derived cell transplantation therapies in the future.
authors.affiliation: Neuroregeneration Laboratories, Harvard Medical School/McLean Hospital, Belmont, Massachusetts, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Sundberg
authors.forename: Helle
authors.initials: H
authors.lastname: Bogetofte
authors.forename: Tristan
authors.initials: T
authors.lastname: Lawson
authors.forename: Johan
authors.initials: J
authors.lastname: Jansson
authors.forename: Gaynor
authors.initials: G
authors.lastname: Smith
authors.forename: Arnar
authors.initials: A
authors.lastname: Astradsson
authors.forename: Michele
authors.initials: M
authors.lastname: Moore
authors.forename: Teresia
authors.initials: T
authors.lastname: Osborn
authors.forename: Oliver
authors.initials: O
authors.lastname: Cooper
authors.forename: Roger
authors.initials: R
authors.lastname: Spealman
authors.forename: Penelope
authors.initials: P
authors.lastname: Hallett
authors.forename: Ole
authors.initials: O
authors.lastname: Isacson
mesh_terms: D000328:Adult; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D004195:Disease Models, Animal; D059290:Dopaminergic Neurons / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation; D005260:Female; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000637:transplantation; D008252:Macaca fascicularis; D008297:Male; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010300:Parkinson Disease / Q000473:pathology / Q000628:therapy*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000637:transplantation; D011897:Random Allocation; D051381:Rats; D033581:Stem Cell Transplantation / Q000379:methods*
keywords: Parkinson's disease; differentiation; embryonic stem cells; flow cytometry; induced pluripotent stem cells; neural differentiation; transplantation; xenotransplantation