@article{da0b4872fd694b998851a4d647c85847,
title = "Generation of GFAP::GFP astrocyte reporter lines from human adult fibroblast-derived iPS cells using zinc-finger nuclease technology",
abstract = "Astrocytes are instrumental to major brain functions, including metabolic support, extracellular ion regulation, the shaping of excitatory signaling events and maintenance of synaptic glutamate homeostasis. Astrocyte dysfunction contributes to numerous developmental, psychiatric and neurodegenerative disorders. The generation of adult human fibroblast-derived induced pluripotent stem cells (iPSCs) has provided novel opportunities to study mechanisms of astrocyte dysfunction in human-derived cells. To overcome the difficulties of cell type heterogeneity during the differentiation process from iPSCs to astroglial cells (iPS astrocytes), we generated homogenous populations of iPS astrocytes using zinc-finger nuclease (ZFN) technology. Enhanced green fluorescent protein (eGFP) driven by the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter was inserted into the safe harbor adeno-associated virus integration site 1 (AAVS1) locus in disease and control-derived iPSCs. Astrocyte populations were enriched using Fluorescence Activated Cell Sorting (FACS) and after enrichment more than 99% of iPS astrocytes expressed mature astrocyte markers including GFAP, S100β, NFIA and ALDH1L1. In addition, mature pure GFP-iPS astrocytes exhibited a well-described functional astrocytic activity in vitro characterized by neuron-dependent regulation of glutamate transporters to regulate extracellular glutamate concentrations. Engraftment of GFP-iPS astrocytes into rat spinal cord grey matter confirmed in vivo cell survival and continued astrocytic maturation. In conclusion, the generation of GFAP::GFP-iPS astrocytes provides a powerful in vitro and in vivo tool for studying astrocyte biology and astrocyte-driven disease pathogenesis and therapy.",
keywords = "Astrocyte, GFAP, IPSC, Induced pluripotent stem cells, Zinc finger nuclease",
author = "Zhang, {Ping Wu} and Haidet-Phillips, {Amanda M.} and Pham, {Jacqueline T.} and Youngjin Lee and Yuqing Huo and Tienari, {Pentti J.} and Maragakis, {Nicholas J.} and Rita Sattler and Rothstein, {Jeffrey D.}",
note = "Funding Information: NIH-NINDS; Grant number: 1U24NS078736, RO1 NS085207 C9, P2ALS, 5U01NS062713; Grant sponsor: Ansari ALS Center for Stem Cell and Regeneration Research (NJM); Grant number: W81XWH-10-1-0520 DOD-ALSRP; Grant sponsors: Brain Science Institute; Target ALS; ALS Association; Muscular Dystrophy Association; The Judith & Jean Pape Adams Charitable Foundation; ALS Association Post-doctoral Fellowship; Muscular Dystrophy Association Career Development Award; The Robert Packard Center for ALS Research at Johns Hopkins; Finnish Academy; Sigrid Juselius Foundation; Helsinki University Central Hospital. The authors thank Hao Zhang, Sara Gross, and Elizabeth Daley for their scientific discussion and technical support. Timo Otonkoski and The Biomedicum Stem Cell Center, University of Helsinki are thanked for providing the resources for making C9orf72 ALS patient{\textquoteright}s iPS cells. Funding Information: NIH-NINDS; Grant number: 1U24NS078736, RO1 NS085207 C9, P2ALS, 5U01NS062713; Grant sponsor: Ansari ALS Center for Stem Cell and Regeneration Research (NJM); Grant number: W81XWH-10-1-0520 DOD-ALSRP; Grant sponsors: Brain Science Institute; Target ALS; ALS Association; Muscular Dystrophy Association; The Judith & Jean Pape Adams Charitable Foundation; ALS Association Post-doctoral Fellowship; Muscular Dystrophy Association Career Development Award; The Robert Packard Center for ALS Research at Johns Hopkins; Finnish Academy; Sigrid Juselius Foundation; Helsinki University Central Hospital. The authors thank Hao Zhang, Sara Gross, and Elizabeth Daley for their scientific discussion and technical support. Timo Otonkoski and The Biomedicum Stem Cell Center, University of Helsinki are thanked for providing the resources for making C9orf72 ALS patient?s iPS cells. Publisher Copyright: {\textcopyright} 2015 Wiley Periodicals, Inc.",
year = "2016",
month = jan,
day = "1",
doi = "10.1002/glia.22903",
language = "English (US)",
volume = "64",
pages = "63--75",
journal = "GLIA",
issn = "0894-1491",
publisher = "John Wiley and Sons Inc.",
number = "1",
}