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- Title
A Targeted NKX2.1 Human Embryonic Stem Cell Reporter Line Enables Identification of Human Basal Forebrain Derivatives.
- Authors
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.
- 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 NKX2.1GFP/w hESCs for investigating human forebrain development and neuronal differentiation. S TEM C ELLS 2011;29:462-473
- Subjects
STEM cells; PLURIPOTENT stem cells; GREEN fluorescent protein; FLUORESCENT polymers; AMINOBUTYRIC acid
- Publication
Stem Cells, 2011, Vol 29, Issue 3, p462
- ISSN
1066-5099
- Publication type
Article
- DOI
10.1002/stem.587