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Efficient generation of mature cerebellar Purkinje cells from mouse embryonic stem cells
Author(s) -
Tao Osamu,
Shimazaki Takuya,
Okada Yohei,
Naka Hayato,
Kohda Kazuhisa,
Yuzaki Michisuke,
Mizusawa Hidehiro,
Okano Hideyuki
Publication year - 2009
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.22208
Subject(s) - cerebellum , synaptogenesis , embryonic stem cell , microbiology and biotechnology , excitatory postsynaptic potential , inhibitory postsynaptic potential , biology , postsynaptic potential , purkinje cell , neuroscience , patch clamp , stem cell , electrophysiology , chemistry , receptor , biochemistry , gene
Mouse embryonic stem cells (ESCs) can generate cerebellar neurons, including Purkinje cells (PCs) and their precursor cells, in a floating 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 Ca 2+ 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. © 2009 Wiley‐Liss, Inc.