Directed differentiation of telencephalic precursors from embryonic stem cells.
We demonstrate directed differentiation of telencephalic precursors from mouse embryonic stem (ES) cells using optimized serum-free suspension culture (SFEB culture). Treatment with Wnt and Nodal antagonists (Dkk1 and LeftyA) during the first 5 d of SFEB culture causes nearly selective neural differentiation in ES cells ( approximately 90%). In the presence of Dkk1, with or without LeftyA, SFEB induces efficient generation ( approximately 35%) of cells expressing telencephalic marker Bf1. Wnt3a treatment during the late culture period increases the pallial telencephalic population (Pax6(+) cells yield up to 75% of Bf1(+) cells), whereas Shh promotes basal telencephalic differentiation (into Nkx2.1(+) and/or Islet1/2(+) cells) at the cost of pallial telencephalic differentiation. Thus, in the absence of caudalizing signals, floating aggregates of ES cells generate naive telencephalic precursors that acquire subregional identities by responding to extracellular patterning signals.
Watanabe
Kiichi
K
Organogenesis and Neurogenesis Group, Center for Developmental Biology, RIKEN, Kobe 650-0047, Japan.
Kamiya
Daisuke
D
Nishiyama
Ayaka
A
Katayama
Tomoko
T
Nozaki
Satoshi
S
Kawasaki
Hiroshi
H
Watanabe
Yasuyoshi
Y
Mizuseki
Kenji
K
Sasai
Yoshiki
Y
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D055457:Nodal Protein / Q000037:antagonists & inhibitors / Q000378:metabolism; D013687:Telencephalon / Q000166:cytology* / Q000196:embryology; D013997:Time Factors; D060509:Wnt3A Protein / Q000037:antagonists & inhibitors / Q000378:metabolism
D017304:Annexin A5; D016895:Culture Media, Serum-Free; D009419:Nerve Tissue Proteins; D055457:Nodal Protein; D060509:Wnt3A Protein