Advertisement

Robust Induction of DARPP32-Expressing GABAergic Striatal Neurons from Human Pluripotent Stem Cells

  • Marija Fjodorova
  • Meng Li
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1780)

Abstract

Efficient generation of disease relevant neuronal subtypes from human pluripotent stem cells (PSCs) is fundamental for realizing their promise in disease modeling, pharmaceutical drug screening and cell therapy. Here we describe a step-by-step protocol for directing the differentiation of human embryonic and induced PSCs (hESCs and hiPSCs, respectively) toward medium spiny neurons, the type of cells that are preferentially lost in Huntington’s disease patients. This method is based on a novel concept of Activin A-dependent induction of the lateral ganglionic/striatal fate using a simple monolayer culture paradigm under chemically defined conditions. Transplantable medium spiny neuron progenitors amenable for cryopreservation are produced in less than 20 days, which differentiate and mature into a high yield of dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDa (DARPP32) expressing gamma-aminobutyric acid (GABA)-ergic neurons in vitro and in the adult rat brain after transplantation. This method has been validated in multiple hESC and hiPSC lines, and is independent of the regime for PSC maintenance.

Keywords

Activin A DARPP32 Lateral ganglionic eminence Medium spiny neuron Pluripotent stem cell Neural differentiation Huntington’s disease Striatum Transplantation 

References

  1. 1.
    Reiner A, Albin RL, Anderson KD et al (1988) Differential loss of striatal projection neurons in Huntington disease. Proc Natl Acad Sci U S A 15:5733–5737CrossRefGoogle Scholar
  2. 2.
    Arber C, Precious SV, Cambray S et al (2015) Activin A directs striatal projection neuron differentiation of human pluripotent stem cells. Development 142:1375–1386CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Feijen A, Goumans MJ, Vandeneijndenvanraaij AJM (1994) Expression of activin subunits, activin receptors and follistatin in postimplantation mouse embryos suggests specific developmental functions for different activins. Development 120:3621–3637PubMedGoogle Scholar
  4. 4.
    Maira M, Long JE, Lee AY et al (2010) Role for TGF-beta superfamily signaling in telencephalic GABAergic neuron development. J Neurodev Disord 2:48–60CrossRefPubMedGoogle Scholar
  5. 5.
    Victor MB, Richner M, Hermanstyne TO et al (2014) Generation of human striatal neurons by microRNA-dependent direct conversion of fibroblasts. Neuron 84:311–323CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Arlotta P, Molyneaux BJ, Jabaudon D et al (2008) Ctip2 controls the differentiation of medium spiny neurons and the establishment of the cellular architecture of the striatum. J Neurosci 28:22–632CrossRefGoogle Scholar
  7. 7.
    Delli Carri A, Onorati M, Lelos MJ et al (2013) Developmentally coordinated extrinsic signals drive human pluripotent stem cell differentiation toward authentic DARPP-32(+) medium-sized spiny neurons. Development 140:301–312CrossRefPubMedGoogle Scholar
  8. 8.
    Nicoleau C, Varela C, Bonnefond C et al (2013) Embryonic stem cells neural differentiation qualifies the role of wnt/beta-catenin signals in human telencephalic specification and regionalization. Stem Cells 31:1763–1774CrossRefPubMedGoogle Scholar
  9. 9.
    Ma L, Hu B, Liu Y et al (2012) Human embryonic stem cell-derived GABA neurons correct locomotion deficits in quinolinic acid-lesioned mice. Cell Stem Cell 10:455–464CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Chambers SM, Fasano CA, Papapetrou EP et al (2009) Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol 27:275–280CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Cambray S, Arber C, Little G et al (2012) Activin induces cortical interneuron identity and differentiation in embryonic stem cell-derived telencephalic neural precursors. Nat Commun 3:841CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Marija Fjodorova
    • 1
  • Meng Li
    • 1
  1. 1.Neuroscience and Mental Health Research Institute, School of BioscienceCardiff UniversityCardiffUK

Personalised recommendations