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Andreas Heuer

Andreas Heuer

Research team manager

Andreas Heuer

Generation of high-purity human ventral midbrain dopaminergic progenitors for in vitro maturation and intracerebral transplantation

Author

  • Sara Nolbrant
  • Andreas Heuer
  • Malin Parmar
  • Agnete Kirkeby

Summary, in English

Generation of precisely patterned neural cells from human pluripotent stem cells (hPSCs) is instrumental in developing disease models and stem cell therapies. Here, we provide a detailed 16-d protocol for obtaining high-purity ventral midbrain (VM) dopamine (DA) progenitors for intracerebral transplantation into animal models and for in vitro maturation into neurons. We have successfully transplanted such cells into the rat; however, in principle, the cells can be used for transplantation into any animal model, and the protocol is designed to also be compatible with clinical transplantation into humans. We show how to precisely set the balance of patterning factors to obtain specifically the caudal VM progenitors that give rise to DA-rich grafts. By specifying how to perform quality control (QC), troubleshooting and adaptation of the procedure, this protocol will facilitate implementation in different laboratories and with a variety of hPSC lines. To facilitate reproducibility of experiments and enable shipping of cells between centers, we present a method for cryopreservation of the progenitors for subsequent direct transplantation or terminal differentiation into DA neurons. This protocol is free of xeno-derived products and can be performed under good manufacturing practice (GMP) conditions.

Department/s

  • StemTherapy: National Initiative on Stem Cells for Regenerative Therapy
  • MultiPark: Multidisciplinary research focused on Parkinson´s disease
  • Developmental and Regenerative Neurobiology
  • Department of Experimental Medical Science
  • Human Neural Developmental Biology

Publishing year

2017-09-01

Language

English

Pages

1962-1979

Publication/Series

Nature Protocols

Volume

12

Issue

9

Document type

Journal article

Publisher

Nature Publishing Group

Topic

  • Neurosciences
  • Cell and Molecular Biology

Status

Published

Research group

  • Developmental and Regenerative Neurobiology
  • Human Neural Developmental Biology

ISBN/ISSN/Other

  • ISSN: 1750-2799