title: A simplified method to generate serotonergic neurons from mouse embryonic stem and induced pluripotent stem cells.
abstract: We have developed a new simple method to induce serotonergic neurons from embryonic stem (ES) and induced pluripotent stem cells. When ES or induced pluripotent stem cells were cultured on a thick gel layer of Matrigel, most colonies extended TuJ1-positive neurites. We found that noggin, a known antagonist of bone morphogenic protein, induces ES cells to express genes involved in serotonergic differentiation, such as Nkx2.2, Pet-1, Sonic hedgehog, tryptophan hydroxylase 2, and serotonin transporter, as well as increases high potassium-induced release of serotonin. To concentrate serotonergic neurons, ES cells carrying Pet-1-enhancer-driven enhanced green fluorescent protein were differentiated and sorted into about 80% pure cultures of serotonergic neurons. Whole cell voltage-clamp recordings showed a voltage-dependent current in dissociated neurons. This simplified method provides an alternative option for serotonergic differentiation of pluripotent stem cells and will likely contribute a deeper understanding regarding the nature of serotonergic neurons and open new therapeutic perspectives for the treatment of psychiatric disorders.
authors.affiliation: Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shimada
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Takai
authors.forename: Kikuko
authors.initials: K
authors.lastname: Shinohara
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamasaki
authors.forename: Keiko
authors.initials: K
authors.lastname: Tominaga-Yoshino
authors.forename: Akihiko
authors.initials: A
authors.lastname: Ogura
authors.forename: Akihiro
authors.initials: A
authors.lastname: Toi
authors.forename: Kouji
authors.initials: K
authors.lastname: Asano
authors.forename: Norihito
authors.initials: N
authors.lastname: Shintani
authors.forename: Atsuko
authors.initials: A
authors.lastname: Hayata-Takano
authors.forename: Akemichi
authors.initials: A
authors.lastname: Baba
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Hashimoto
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D003094:Collagen / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004558:Electric Stimulation; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000502:physiology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics; D006624:Hippocampus / Q000166:cytology; D000090322:Homeobox Protein Nkx-2.2; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D007797:Laminin / Q000378:metabolism; D051379:Mice; D009924:Organ Culture Techniques; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D022661:RNA, Untranslated; D059326:Serotonergic Neurons / Q000502:physiology*; D012701:Serotonin / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D014161:Transduction, Genetic; D014404:Tubulin / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D000099176:Noggin Protein
chemical_list: D055415:Bone Morphogenetic Protein 4; D002352:Carrier Proteins; D003094:Collagen; D004338:Drug Combinations; D049452:Green Fluorescent Proteins; D000090322:Homeobox Protein Nkx-2.2; D007797:Laminin; D011506:Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; D022661:RNA, Untranslated; D012701:Serotonin; D014157:Transcription Factors; D014404:Tubulin; D014446:Tyrosine 3-Monooxygenase; D000099176:Noggin Protein; C525810:Bmp4 protein, mouse; C484684:Fev protein, mouse; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; C000715627:Nkx2-2 protein, mouse; C462941:beta3 tubulin, mouse; C485184:enhanced green fluorescent protein; C059485:matrigel