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In vitro and in vivo analyses of human embryonic stem cell-derived dopamine neurons

Cited 209 time in Web of Science Cited 233 time in Scopus
Authors
Park, Chang-Hwan; Minn, Yang-Ki; Lee, Ji-Yeon; Choi, Dong Ho; Chang, Mi-Yoon; Shim, Jae-Won; Ko, Ji-Yun; Koh, Hyun-Chul; Kang, Min Jeong; Kang, Jin Sun; Rhie, Duck-Joo; Lee, Yong-Sung; Son, Hyeon; Moon, Shin Yong; Kim, Kwang-Soo; Lee, Sang-Hun
Issue Date
2005-02-18
Publisher
Wiley-Blackwell
Citation
J Neurochem. 2005 Mar;92(5):1265-76.
Keywords
AnimalsBehavior, AnimalCalcium-Binding Protein, Vitamin D-Dependent/metabolismCell Differentiation/drug effects/*physiologyCells, CulturedChromatography, High Pressure Liquid/methodsCoculture Techniques/methodsDNA-Binding Proteins/genetics/metabolismDopamine/*metabolism*Embryonic InductionFetusFibroblast Growth Factor 8Fibroblast Growth Factors/pharmacologyGene Expression Regulation, Developmental/physiologyGlial Fibrillary Acidic Protein/metabolismHN Protein/genetics/metabolismHedgehog ProteinsHomeodomain Proteins/genetics/metabolismHumansImmunohistochemistry/methodsIndoles/diagnostic useIntermediate Filament Proteins/metabolismIsotonic Solutions/pharmacologyKi-67 Antigen/metabolismMaleMembrane Potentials/physiologyMesencephalon/cytology/embryologyMicrotubule-Associated Proteins/metabolismNerve Tissue Proteins/metabolismNeurons/*metabolismOrganic Cation Transport Proteins/genetics/metabolismPAX2 Transcription FactorPatch-Clamp Techniques/methodsPotassium Chloride/pharmacologyProliferating Cell Nuclear Antigen/metabolismRNA, Messenger/biosynthesisRatsRats, Sprague-DawleyReverse Transcriptase Polymerase Chain Reaction/methodsRotationStem Cell Transplantation/methodsStem Cells/cytology/*physiologyStromal Cells/physiologyTime FactorsTrans-Activators/pharmacologyTranscription Factors/genetics/metabolismTubulin/metabolismTyrosine 3-Monooxygenase/genetics/metabolismVimentin/metabolismalpha-Fetoproteins/metabolismgamma-Aminobutyric Acid/metabolism
Abstract
Human embryonic stem (hES) cells, due to their capacity of multipotency and self-renewal, may serve as a valuable experimental tool for human developmental biology and may provide an unlimited cell source for cell replacement therapy. The purpose of this study was to assess the developmental potential of hES cells to replace the selectively lost midbrain dopamine (DA) neurons in Parkinson's disease. Here, we report the development of an in vitro differentiation protocol to derive an enriched population of midbrain DA neurons from hES cells. Neural induction of hES cells co-cultured with stromal cells, followed by expansion of the resulting neural precursor cells, efficiently generated DA neurons with concomitant expression of transcriptional factors related to midbrain DA development, such as Pax2, En1 (Engrailed-1), Nurr1, and Lmx1b. Using our procedure, the majority of differentiated hES cells (> 95%) contained neuronal or neural precursor markers and a high percentage (> 40%) of TuJ1+ neurons was tyrosine hydroxylase (TH)+, while none of them expressed the undifferentiated ES cell marker, Oct 3/4. Furthermore, hES cell-derived DA neurons demonstrated functionality in vitro, releasing DA in response to KCl-induced depolarization and reuptake of DA. Finally, transplantation of hES-derived DA neurons into the striatum of hemi-parkinsonian rats failed to result in improvement of their behavioral deficits as determined by amphetamine-induced rotation and step-adjustment. Immunohistochemical analyses of grafted brains revealed that abundant hES-derived cells (human nuclei+ cells) survived in the grafts, but none of them were TH+. Therefore, unlike those from mouse ES cells, hES cell-derived DA neurons either do not survive or their DA phenotype is unstable when grafted into rodent brains.
ISSN
0022-3042 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15715675

http://hdl.handle.net/10371/11886
DOI
https://doi.org/10.1111/j.1471-4159.2004.03006.x
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College of Medicine/School of Medicine (의과대학/대학원)Obstetrics & Gynecology (산부인과전공)Journal Papers (저널논문_산부인과학전공)
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