title: Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies.
abstract: In vitro neural differentiation of human embryonic stem cells (hESCs) is an advantageous system for studying early neural development. The process of early neural differentiation in hESCs begins by initiation of primitive neuroectoderm, which is manifested by rosette formation, with consecutive differentiation into neural progenitors and early glial-like cells. In this study, we examined the involvement of early neural markers - OTX2, PAX6, Sox1, Nestin, NR2F1, NR2F2, and IRX2 - in the onset of rosette formation, during spontaneous neural differentiation of hESC and human induced pluripotent stem cell (hiPSC) colonies. This is in contrast to the conventional way of studying rosette formation, which involves induction of neuronal differentiation and the utilization of embryoid bodies. Here we show that OTX2 is highly expressed at the onset of rosette formation, when rosettes comprise no more than 3-5 cells, and that its expression precedes that of established markers of early neuronal differentiation. Importantly, the rise of OTX2 expression in these cells coincides with the down-regulation of the pluripotency marker OCT4. Lastly, we show that cells derived from rosettes that emerge during spontaneous differentiation of hESCs or hiPSCs are capable of differentiating into dopaminergic neurons in vitro, and into mature-appearing pyramidal and serotonergic neurons weeks after being injected into the motor cortex of NOD-SCID mice.
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: s-malchenko@northwestern.edu.
authors: Sergey
authors: S
authors: Malchenko
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: JXie@luriechildrens.org.
authors: Jianping
authors: J
authors: Xie
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: mbonaldo@luriechildrens.org.
authors: Maria
authors: M
authors: de Fatima Bonaldo
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: EVanin@luriechildrens.org.
authors: Elio F
authors: EF
authors: Vanin
authors: Molecular Pharmacology & Biological Chemistry, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA. Electronic address: bula@northwestern.edu.
authors: Bula J
authors: BJ
authors: Bhattacharyya
authors: Molecular Pharmacology & Biological Chemistry, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA. Electronic address: a-belmadni@northwestern.edu.
authors: Abdelhak
authors: A
authors: Belmadani
authors: Division of Pediatric Neurosurgery, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: gxi@luriechildrens.org.
authors: Guifa
authors: G
authors: Xi
authors: Developmental Biology Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA. Electronic address: v-galat@northwestern.edu.
authors: Vasily
authors: V
authors: Galat
authors: Microscopy and Imaging Facility, Ann & Robert H. Lurie Children's Hospital of Chicago Research Center, Chicago, IL, USA. Electronic address: wgoossens@luriechildrens.org.
authors: William
authors: W
authors: Goossens
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Electronic address: r-seftor@northwestern.edu.
authors: Richard E B
authors: RE
authors: Seftor
authors: Division of Pediatric Neurosurgery, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA. Electronic address: ttomita@luriechildrens.org.
authors: Tadanori
authors: T
authors: Tomita
authors: Division of Hematology/Oncology, Department of Pediatrics, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA; Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Electronic address: j-crispino@northwestern.edu.
authors: John
authors: J
authors: Crispino
authors: Molecular Pharmacology & Biological Chemistry, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA. Electronic address: r-miller10@northwestern.edu.
authors: Richard J
authors: RJ
authors: Miller
authors: Neurobiology Program, Ann & Robert H. Lurie Children's Hospital of Chicago Research Center, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA. Electronic address: m-bohn@northwestern.edu.
authors: Martha C
authors: MC
authors: Bohn
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA; Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Electronic address: m-hendrix@northwestern.edu.
authors: Mary J C
authors: MJ
authors: Hendrix
authors: Cancer Biology and Epigenomics Program, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA; Division of Hematology/Oncology, Department of Pediatrics, Northwestern University, Feinberg School of Medicine, Chicago, IL, USA; Falk Brain Tumor Center, Ann & Robert H Lurie Children's Hospital of Chicago Research Center, Ann & Robert H Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA; Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Electronic address: m-soares@northwestern.edu.
authors: Marcelo B
authors: MB
authors: Soares
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D059290:Dopaminergic Neurons / Q000378:metabolism / Q000502:physiology*; D015536:Down-Regulation / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009044:Motor Cortex / Q000378:metabolism / Q000502:physiology; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D051857:Otx Transcription Factors / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D012397:Rosette Formation / Q000379:methods*; D059326:Serotonergic Neurons / Q000378:metabolism / Q000502:physiology*
chemical_list: D015415:Biomarkers; C497659:OTX2 protein, human; D050814:Octamer Transcription Factor-3; D051857:Otx Transcription Factors; C494173:POU5F1 protein, human
keywords: ACSF; BLBP; EB; GFAP; GFP; Induced pluripotent stem cells; Mature pyramidal neurons; NEC; NOD-SCID; NP; Neural rosettes; RG; RNSC; Radial glia; Rosette neural stem cell line; TH; artificial celebrospinal fluid; brain-lipid-binding protein; embryoid bodies; glial fibrillary acidic protein; green fluorescent protein; hESCs; hiPSC; human embryonic stem cells; human induced pluripotent stem cell; neural progenitors; non-obese diabetes-severe combined immunodeficiency; primitive neuroectoderm; radial glia; tyrosine hydroxylase