Cryopreservation Maintains Functionality of Human iPSC Dopamine Neurons and Rescues Parkinsonian Phenotypes In Vivo

Stem Cell Reports. 2017 Jul 11;9(1):149-161. doi: 10.1016/j.stemcr.2017.04.033. Epub 2017 Jun 1.

Abstract

A major challenge for clinical application of pluripotent stem cell therapy for Parkinson's disease (PD) is large-scale manufacturing and cryopreservation of neurons that can be efficiently prepared with minimal manipulation. To address this obstacle, midbrain dopamine neurons were derived from human induced pluripotent stem cells (iPSC-mDA) and cryopreserved in large production lots for biochemical and transplantation studies. Cryopreserved, post-mitotic iPSC-mDA neurons retained high viability with gene, protein, and electrophysiological signatures consistent with midbrain floor-plate lineage. To test therapeutic efficacy, cryopreserved iPSC-mDA neurons were transplanted without subculturing into the 6-OHDA-lesioned rat and MPTP-lesioned non-human-primate models of PD. Grafted neurons retained midbrain lineage with extensive fiber innervation in both rodents and monkeys. Behavioral assessment in 6-OHDA-lesioned rats demonstrated significant reversal in functional deficits up to 6 months post transplantation with reinnervation of the host striatum and no aberrant growth, supporting the translational development of pluripotent cell-based therapies in PD.

Keywords: 6-OHDA; MPTP; Parkinson's disease; cryopreservation; iPSC; midbrain dopamine neuron; non-human primate.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Corpus Striatum / cytology
  • Corpus Striatum / pathology
  • Cryopreservation* / methods
  • Disease Models, Animal
  • Dopamine / metabolism
  • Dopaminergic Neurons / cytology*
  • Dopaminergic Neurons / metabolism
  • Dopaminergic Neurons / transplantation*
  • Haplorhini
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Mesencephalon / cytology
  • Mesencephalon / pathology
  • Neurogenesis
  • Parkinson Disease / pathology
  • Parkinson Disease / therapy*
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Dopamine