title: Selective TGF-β1/ALK inhibitor improves neuronal differentiation of mouse embryonic stem cells.
abstract: The transforming growth factor-β1 (TGF-β1), a polypeptide member of the TGF-β superfamily, has myriad cellular functions, including cell fate differentiation. We hypothesized that suppression of TGF-β1 signaling would improve the efficacy of neuronal differentiation during embryoid body (EB) development. In this study, mouse embryonic stem cells (ESCs) were allowed to differentiate into their neuronal lineage, both with, and without the TGF-β1 inhibitor (A83-01). After 8 days of EB suspension culture, the samples were examined by morphological analysis, immunocytochemistry and immunohistochemistry with pluripotent (Oct4, Sox2) and neuronal specific markers (Pax6, NeuN). The alteration of gene expressions during EB development was determined by quantitative RT-PCR. Our results revealed that the TGF-β1/ALK inhibitor potentially suppressed pluripotent gene (Oct4) during a rapidly up-regulation of neuronal associated genes including Sox1 and MAP2. Strikingly, during EB development, the expression of GFAP, the astrocyte specific gene, remarkably decreased compared to the non-treated control. This strategy demonstrated the beneficial function of TGF-β1/ALK inhibitor that rapidly and uniformly drives cell fate alteration from pluripotent state toward neuronal lineages.
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand; Siriraj Center of Excellence for Stem Cell Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. Electronic address: k.nuttha@gmail.com.
authors.forename: Nuttha
authors.initials: N
authors.lastname: Klincumhom
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand. Electronic address: tharasanit@hotmail.com.
authors.forename: Theerawat
authors.initials: T
authors.lastname: Tharasanit
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand. Electronic address: giftgold10@hotmail.com.
authors.forename: Chommanart
authors.initials: C
authors.lastname: Thongkittidilok
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand. Electronic address: yurikakarin_2004@hotmail.com.
authors.forename: Narong
authors.initials: N
authors.lastname: Tiptanavattana
authors.affiliation: Department of Preclinical and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand. Electronic address: sasitorn.run@mahidol.edu.
authors.forename: Sasitorn
authors.initials: S
authors.lastname: Rungarunlert
authors.affiliation: Biotalentum Ltd., Aulich Lajos u. 26, 2100 Gödöllő, Hungary; Molecular Animal Biotechnology Laboratory, Szent Istvan University, 2100 Gödöllő, Hungary; Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, 3584 CL Utrecht, The Netherlands. Electronic address: andras.dinnyes@biotalentum.hu.
authors.forename: András
authors.initials: A
authors.lastname: Dinnyés
authors.affiliation: Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand. Electronic address: tmongkol@chula.ac.th.
authors.forename: Mongkol
authors.initials: M
authors.lastname: Techakumphu
mesh_terms: D030201:Activin Receptors, Type I / Q000037:antagonists & inhibitors; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D058732:Embryoid Bodies; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D051379:Mice; D058953:Neural Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D055495:Neurogenesis / Q000187:drug effects*; D009457:Neuroglia / Q000378:metabolism / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D011720:Pyrazoles / Q000494:pharmacology*; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D015398:Signal Transduction; D013859:Thiocarbamates / Q000494:pharmacology*; D013882:Thiosemicarbazones; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: C507011:A-83-01; D011720:Pyrazoles; D018125:Receptors, Transforming Growth Factor beta; D013859:Thiocarbamates; D013882:Thiosemicarbazones; D053773:Transforming Growth Factor beta1; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I
keywords: Embryonic stem cells; Mouse; Neuronal differentiation; TGF-β1/ALK inhibitor