A Targeted NKX2.1 Human Embryonic Stem Cell Reporter Line Enables Identification of Human Basal Forebrain Derivatives

作者：Goulburn Adam L; Alden Darym; Davis Richard P; Micallef Suzanne J; Ng Elizabeth S; Yu Qing C; Lim Sue Mei; Soh Chew Li; Elliott David A; Hatzistavrou Tanya; Bourke Justin; Watmuff Bradley; Lang Richard J; Haynes John M; Pouton Colin W; Giudice Antonietta; Trounson Alan O; Anderson Stewart A; Stanley Edouard G; Elefanty Andrew G^*

来源：Stem Cells, 2011, 29(3): 462-473.

DOI：10.1002/stem.587

• Human embryonic stem cells
• NKX2.1
• FGF
• Ventral telencephalon
• Hypothalamus
• Retinoic acid
• MGE

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 gamma-aminobutyric acid-, tyrosine hydroxylase-, and somatostatin-expressing neurons as well as to platelet-derived growth factor receptor a-positive oligodendrocyte precursors. These studies highlight the diversity of cell types that can be generated from human NKX2.1 1 progenitors and demonstrate the utility of NKX2.1(GFP/w) hESCs for investigating human forebrain development and neuronal differentiation. STEM CELLS 2011;29:462-473

• 出版日期2011-3