Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease

This article has been updated


Human pluripotent stem cells (PSCs) are a promising source of cells for applications in regenerative medicine. Directed differentiation of PSCs into specialized cells such as spinal motoneurons1 or midbrain dopamine (DA) neurons2 has been achieved. However, the effective use of PSCs for cell therapy has lagged behind. Whereas mouse PSC-derived DA neurons have shown efficacy in models of Parkinson’s disease3,4, DA neurons from human PSCs generally show poor in vivo performance5. There are also considerable safety concerns for PSCs related to their potential for teratoma formation or neural overgrowth6,7. Here we present a novel floor-plate-based strategy for the derivation of human DA neurons that efficiently engraft in vivo, suggesting that past failures were due to incomplete specification rather than a specific vulnerability of the cells. Midbrain floor-plate precursors are derived from PSCs 11 days after exposure to small molecule activators of sonic hedgehog (SHH) and canonical WNT signalling. Engraftable midbrain DA neurons are obtained by day 25 and can be maintained in vitro for several months. Extensive molecular profiling, biochemical and electrophysiological data define developmental progression and confirm identity of PSC-derived midbrain DA neurons. In vivo survival and function is demonstrated in Parkinson’s disease models using three host species. Long-term engraftment in 6-hydroxy-dopamine-lesioned mice and rats demonstrates robust survival of midbrain DA neurons derived from human embryonic stem (ES) cells, complete restoration of amphetamine-induced rotation behaviour and improvements in tests of forelimb use and akinesia. Finally, scalability is demonstrated by transplantation into parkinsonian monkeys. Excellent DA neuron survival, function and lack of neural overgrowth in the three animal models indicate promise for the development of cell-based therapies in Parkinson’s disease.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Induction and neurogenic conversion of ES-cell-derived midbrain FP precursors is dependent on CHIR addition.
Figure 2: Immunocytochemical and molecular analysis of midbrain DA neuron fate in LSB/S/F8/CHIR-treated versus LSB/S/F8 (hypothalamic) and forebrain LSB (dorsal forebrain) fates.
Figure 3: In vitro maturation and functional characterization of FP versus rosette-derived midbrain DA neurons.
Figure 4: In vivo survival and function of FP-derived human DA neurons in mouse, rat and monkey Parkinson’s disease hosts.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

The raw gene expression data generated in this study have been deposited in the Gene Expression Omnibus database under accession number GSE32658.

Change history

  • 21 December 2011

    In the online-only Methods, in the "Neural induction" subsection, the concentration of DAPT appeared incorrectly as "10 nM" in the AOP PDF version. This has been corrected.


  1. 1

    Li, X. J. et al. Specification of motoneurons from human embryonic stem cells. Nature Biotechnol. 23, 215–221 (2005)

    Article  Google Scholar 

  2. 2

    Perrier, A. L. et al. From the cover: derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl Acad. Sci. USA 101, 12543–12548 (2004)

    ADS  CAS  Article  Google Scholar 

  3. 3

    Tabar, V. et al. Therapeutic cloning in individual parkinsonian mice. Nature Med. 14, 379–381 (2008)

    CAS  Article  Google Scholar 

  4. 4

    Wernig, M. et al. Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson’s disease. Proc. Natl Acad. Sci. USA 105, 5856–5861 (2008)

    ADS  CAS  Article  Google Scholar 

  5. 5

    Lindvall, O. & Kokaia, Z. Stem cells in human neurodegenerative disorders–time for clinical translation? J. Clin. Invest. 120, 29–40 (2010)

    CAS  Article  Google Scholar 

  6. 6

    Elkabetz, Y. et al. Human ES cell-derived neural rosettes reveal a functionally distinct early neural stem cell stage. Genes Dev. 22, 152–165 (2008)

    CAS  Article  Google Scholar 

  7. 7

    Roy, N. S. et al. Functional engraftment of human ES cell-derived dopaminergic neurons enriched by coculture with telomerase-immortalized midbrain astrocytes. Nature Med. 12, 1259–1268 (2006)

    CAS  Article  Google Scholar 

  8. 8

    Kittappa, R., Chang, W. W., Awatramani, R. B. & McKay, R. D. The foxa2 gene controls the birth and spontaneous degeneration of dopamine neurons in old age. PLoS Biol. 5, e325 (2007)

    Article  Google Scholar 

  9. 9

    Ferri, A. L. et al. Foxa1 and Foxa2 regulate multiple phases of midbrain dopaminergic neuron development in a dosage-dependent manner. Development 134, 2761–2769 (2007)

    CAS  Article  Google Scholar 

  10. 10

    Roelink, H. et al. Floor plate and motor neuron induction by vhh-1, a vertebrate homolog of hedgehog expressed by the notochord. Cell 76, 761–775 (1994)

    CAS  Article  Google Scholar 

  11. 11

    Liem, K. F., Tremml, G., Roelink, H. & Jessell, T. M. Dorsal differentiation of neural plate cells induced by BMP-mediated signals from epidermal ectoderm. Cell 82, 969–979 (1995)

    CAS  Article  Google Scholar 

  12. 12

    Fasano, C. A., Chambers, S. M., Lee, G., Tomishima, M. J. & Studer, L. Efficient derivation of functional floor plate tissue from human embryonic stem cells. Cell Stem Cell 6, 336–347 (2010)

    CAS  Article  Google Scholar 

  13. 13

    Chambers, S. M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature Biotechnol. 27, 275–280 (2009)

    CAS  Article  Google Scholar 

  14. 14

    Muroyama, Y., Fujihara, M., Ikeya, M., Kondoh, H. & Takada, S. Wnt signaling plays an essential role in neuronal specification of the dorsal spinal cord. Genes Dev. 16, 548–553 (2002)

    CAS  Article  Google Scholar 

  15. 15

    Joksimovic, M. et al. Wnt antagonism of Shh facilitates midbrain floor plate neurogenesis. Nature Neurosci. 12, 125–131 (2009)

    CAS  Article  Google Scholar 

  16. 16

    Lyashenko, N. et al. Differential requirement for the dual functions of beta-catenin in embryonic stem cell self-renewal and germ layer formation. Nature Cell Biol. 13, 753–761 (2011)

    CAS  Article  Google Scholar 

  17. 17

    VanDunk, C., Hunter, L. A. & Gray, P. A. Development, maturation, and necessity of transcription factors in the mouse suprachiasmatic nucleus. J. Neurosci. 31, 6457–6467 (2011)

    CAS  Article  Google Scholar 

  18. 18

    Huang, W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nature Protocols 4, 44–57 (2009)

    CAS  Article  Google Scholar 

  19. 19

    Costa, R. H., Grayson, D. R. & Darnell, J. E., Jr Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes. Mol. Cell. Biol. 9, 1415–1425 (1989)

    CAS  Article  Google Scholar 

  20. 20

    Soldner, F. et al. Parkinson’s disease patient-derived induced pluripotent stem cells free of viral reprogramming factors. Cell 136, 964–977 (2009)

    CAS  Article  Google Scholar 

  21. 21

    Guzman, J. N., Sanchez-Padilla, J., Chan, C. S. & Surmeier, D. J. Robust pacemaking in substantia nigra dopaminergic neurons. J. Neurosci. 29, 11011–11019 (2009)

    CAS  Article  Google Scholar 

  22. 22

    Nedergaard, S., Flatman, J. A. & Engberg, I. Nifedipine- and omega-conotoxin-sensitive Ca2+ conductances in guinea-pig substantia nigra pars compacta neurones. J. Physiol. (Lond.) 466, 727–747 (1993)

    CAS  Google Scholar 

  23. 23

    Ferrari, D., Sanchez-Pernaute, R., Lee, H., Studer, L. & Isacson, O. Transplanted dopamine neurons derived from primate ES cells preferentially innervate DARPP-32 striatal progenitors within the graft. Eur. J. Neurosci. 24, 1885–1896 (2006)

    Article  Google Scholar 

  24. 24

    Olanow, C. W., Kordower, J. H. & Freeman, T. B. Fetal nigral transplantation as a therapy for Parkinson’s disease. Trends Neurosci. 19, 102–109 (1996)

    CAS  Article  Google Scholar 

  25. 25

    Zetterström, R. H. et al. Dopamine neuron agenesis in Nurr1-deficient mice. Science 276, 248–250 (1997)

    Article  Google Scholar 

  26. 26

    Quintana, E. et al. Efficient tumour formation by single human melanoma cells. Nature 456, 593–598 (2008)

    ADS  CAS  Article  Google Scholar 

  27. 27

    Kim, H. et al. miR-371-3 expression predicts neural differentiation propensity in human pluripotent stem cells. Cell Stem Cell 8, 695–706 (2011)

    CAS  Article  Google Scholar 

  28. 28

    Hargus, G. et al. Differentiated Parkinson patient-derived induced pluripotent stem cells grow in the adult rodent brain and reduce motor asymmetry in Parkinsonian rats. Proc. Natl Acad. Sci. USA 107, 15921–15926 (2010)

    ADS  CAS  Article  Google Scholar 

  29. 29

    Aubry, L. et al. Striatal progenitors derived from human ES cells mature into DARPP32 neurons in vitro and in quinolinic acid-lesioned rats. Proc. Natl Acad. Sci. USA 105, 16707–16712 (2008)

    ADS  CAS  Article  Google Scholar 

  30. 30

    Ban, H. et al. Efficient generation of transgene-free human induced pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors. Proc. Natl. Acad. Sci. USA 108, 14234–14239 (2011)

    ADS  CAS  Article  Google Scholar 

  31. 31

    Studer, L., Tabar, V. & McKay, R. D. Transplantation of expanded mesencephalic precursors leads to recovery in parkinsonian rats. Nature Neurosci. 1, 290–295 (1998)

    CAS  Article  Google Scholar 

  32. 32

    Kordower, J. H. et al. Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson’s disease. Science 290, 767–773 (2000)

    ADS  CAS  Article  Google Scholar 

  33. 33

    Paxinos, G., Huang, X.-F. & Toga, A. W. The Rhesus Monkey Brain in Stereotaxic Coordinates (Academic Press, 2000)

    Google Scholar 

  34. 34

    Blume, S. R., Cass, D. K. & Tseng, K. Y. Stepping test in mice: a reliable approach in determining forelimb akinesia in MPTP-induced Parkinsonism. Exp. Neurol. 219, 208–211 (2009)

    Article  Google Scholar 

  35. 35

    Crawley, J. N. What’s Wrong With My Mouse: Behavioral Phenotyping of Transgenic and Knockout Mice (Wiley, 2000)

    Google Scholar 

  36. 36

    Studer, L. et al. Noninvasive dopamine determination by reversed phase HPLC in the medium of free-floating roller tube cultures of rat fetal ventral mesencephalon: A tool to assess dopaminergic tissue prior to grafting. Brain Res. Bull. 41, 143–150 (1996)

    CAS  Article  Google Scholar 

  37. 37

    Tabar, V. et al. Migration and differentiation of neural precursors derived from human embryonic stem cells in the rat brain. Nature Biotechnol. 23, 601–606 (2005)

    CAS  Article  Google Scholar 

Download references


We thank K. Manova, M. Tomishima and A. Viale for excellent technical support, and R. McKay for the anti-nestin antibody. The work was supported by NIH/NINDS grant NS052671, the European Commission project NeuroStemcell, the Starr foundation and NYSTEM contract C024414 to L.S.; by NYSTEM contract C024413, the Michael T. McCarthy Foundation and the Elkus Family Foundation to V.T.; by the Consolidated Anti-Aging Foundation to J.H.K.; by NIH/NINDS grant P50 NS047085, and support from Falk Medical Research Trust to D.J.S.; J.-W.S. was supported by NYSCF (Druckenmiller fellowship) and S.K. by a Starr stem cell scholar fellowship.

Author information




S.K. and J.-W.S.: conception and study design, maintenance and directed differentiation of PSCs, cellular/molecular assays, histological analyses, mouse behavioural assays, data interpretation and writing of manuscript. J.P., G.A., C.A. and A.B.: rat transplantation, histological analyses and behavioural assays. Y.M.G.: mouse transplantation and histological analyses. D.R.W. and J.H.K.: monkey transplantation, histological analysis and data interpretation. L.Y. and M.F.B.: HPLC analysis and data interpretation. L.C.-R., Z.X and D.J.S.: electrophysiological analyses and data interpretation. V.T.: study design, data analysis and writing of manuscript. L.S.: conception and study design, data analysis and interpretation, and writing of manuscript.

Corresponding author

Correspondence to Lorenz Studer.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-11 with legends and Supplementary Tables 1-6.s (PDF 0 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kriks, S., Shim, JW., Piao, J. et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature 480, 547–551 (2011).

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing