title: A targeted NKX2.1 human embryonic stem cell reporter line enables identification of human basal forebrain derivatives.
abstract: We have used homologous recombination in human embryonic stem cells (hESCs) to insert sequences encoding green fluorescent protein (GFP) into the NKX2.1 locus, a gene required for normal development of the basal forebrain. Generation of NKX2.1-GFP(+) cells was dependent on the concentration, timing, and duration of retinoic acid treatment during differentiation. NKX2.1-GFP(+) progenitors expressed genes characteristic of the basal forebrain, including SHH, DLX1, LHX6, and OLIG2. Time course analysis revealed that NKX2.1-GFP(+) cells could upregulate FOXG1 expression, implying the existence of a novel pathway for the generation of telencephalic neural derivatives. Further maturation of NKX2.1-GFP(+) cells gave rise to γ-aminobutyric acid-, tyrosine hydroxylase-, and somatostatin-expressing neurons as well as to platelet-derived growth factor receptor α-positive oligodendrocyte precursors. These studies highlight the diversity of cell types that can be generated from human NKX2.1(+) progenitors and demonstrate the utility of NKX2.1(GFP/w) hESCs for investigating human forebrain development and neuronal differentiation.
authors.affiliation: Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Victoria, Australia.
authors.forename: Adam L
authors.initials: AL
authors.lastname: Goulburn
authors.forename: Darym
authors.initials: D
authors.lastname: Alden
authors.forename: Richard P
authors.initials: RP
authors.lastname: Davis
authors.forename: Suzanne J
authors.initials: SJ
authors.lastname: Micallef
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: Ng
authors.forename: Qing C
authors.initials: QC
authors.lastname: Yu
authors.forename: Sue Mei
authors.initials: SM
authors.lastname: Lim
authors.forename: Chew-Li
authors.initials: CL
authors.lastname: Soh
authors.forename: David A
authors.initials: DA
authors.lastname: Elliott
authors.forename: Tanya
authors.initials: T
authors.lastname: Hatzistavrou
authors.forename: Justin
authors.initials: J
authors.lastname: Bourke
authors.forename: Bradley
authors.initials: B
authors.lastname: Watmuff
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Lang
authors.forename: John M
authors.initials: JM
authors.lastname: Haynes
authors.forename: Colin W
authors.initials: CW
authors.lastname: Pouton
authors.forename: Antonietta
authors.initials: A
authors.lastname: Giudice
authors.forename: Alan O
authors.initials: AO
authors.lastname: Trounson
authors.forename: Stewart A
authors.initials: SA
authors.lastname: Anderson
authors.forename: Edouard G
authors.initials: EG
authors.lastname: Stanley
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Elefanty
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D019070:Cell Lineage / Q000235:genetics*; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D017930:Genes, Reporter; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D058990:Molecular Targeted Therapy / Q000379:methods; D055495:Neurogenesis / Q000235:genetics / Q000502:physiology; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology / Q000196:embryology* / Q000502:physiology; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: C000614414:Nkx2-1 protein, mouse; D009687:Nuclear Proteins; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors