A Targeted NKX2.1 Human Embryonic Stem Cell Reporter Line Enables Identification of Human Basal Forebrain Derivatives
Adam L Goulburn, Darym Alden, Richard P Davis, Suzanne J Micallef, Elizabeth S Ng, Qing C Yu, Sue Mei Lim, Chew-Li Soh, David A Elliott, Tanya Hatzistavrou, Justin Bourke, Bradley Watmuff, Richard J Lang, John M Haynes, Colin W Pouton, Antonietta Giudice, Alan O Trounson, Stewart A Anderson, Edouard G Stanley, Andrew G Elefanty
STEM CELLS | WILEY | Published : 2011
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..View full abstract
We thank Robyn Mayberry and Amanda Bruce for the provision of hESCs, FlowCore for flow cytometric sorting, Monash Micro Imaging for confocal microscopy and time-lapse video services. This work was supported by the Australian Stem Cell Centre (ASCC), The National Health and Medical Research Council (NHMRC) of Australia, The Juvenile Diabetes Research Foundation and the Starr Foundation (D.A. and S.A.A.). A.G.E. and E.G.S. are Senior Research Fellows of the NHMRC. R.P.D. is currently affiliated with Department of Anatomy and Embryology, Leiden University Medical Centre, Leiden, The Netherlands.