Efficient generation of mature cerebellar Purkinje cells from mouse embryonic stem cells

Osamu Tao, Takuya Shimazaki, Yohei Okada, Hayato Naka, Kazuhisa Kohda, Michisuke Yuzaki, Hidehiro Mizusawa, Hideyuki Okano

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

Mouse embryonic stem cells (ESCs) can generate cerebellar neurons, including Purkinje cells (PCs) and their precursor cells, in a .oating culture system called serum-free culture of embryoid body-like aggregates (SFEB) treated with BMP4, Fgf8b, and Wnt3a. Here we successfully established a coculture system that induced the maturation of PCs in ESC-derived Purkinje cell (EDPC) precursors in SFEB, using as a feeder layer a cerebellum dissociation culture prepared from mice at postnatal day (P) 6-8. PC maturation was incomplete or abnormal when the adherent culture did not include feeder cells or when the feeder layer was from neonatal cerebellum. In contrast, EDPCs exhibited the morphology of mature PCs and synaptogenesis with other cerebellar neurons when grown for 4 weeks in coculture system with the postnatal cerebellar feeder. Furthermore, the electrophysiological properties of these EDPCs were compatible with those of native mature PCs in vitro, such as Na+ or Ca2 + spikes elicited by current injections and excitatory or inhibitory postsynaptic currents, which were assessed by whole-cell patch-clamp recordings. Thus, EDPC precursors in SFEB can mature into PCs whose properties are comparable with those of native PCs in vitro.

Original languageEnglish
Pages (from-to)234-247
Number of pages14
JournalJournal of neuroscience research
Volume88
Issue number2
DOIs
Publication statusPublished - 2010 Feb 1

Keywords

  • Cerebellar Purkinje cells
  • Coculture
  • Mouse embryonic stem cells
  • SFEB

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

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