{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Sluch VM"],"funding":["NEI NIH HHS","PHS HHS"],"pagination":["16595"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC4643248"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["5"],"pubmed_abstract":["Retinal ganglion cell (RGC) injury and cell death from glaucoma and other forms of optic nerve disease is a major cause of irreversible vision loss and blindness. Human pluripotent stem cell (hPSC)-derived RGCs could provide a source of cells for the development of novel therapeutic molecules as well as for potential cell-based therapies. In addition, such cells could provide insights into human RGC development, gene regulation, and neuronal biology. Here, we report a simple, adherent cell culture protocol for differentiation of hPSCs to RGCs using a CRISPR-engineered RGC fluorescent reporter stem cell line. Fluorescence-activated cell sorting of the differentiated cultures yields a highly purified population of cells that express a range of RGC-enriched markers and exhibit morphological and physiological properties typical of RGCs. Additionally, we demonstrate that aligned nanofiber matrices can be used to guide the axonal outgrowth of hPSC-derived RGCs for in vitro optic nerve-like modeling. Lastly, using this protocol we identified forskolin as a potent promoter of RGC differentiation."],"journal":["Scientific reports"],"pubmed_title":["Differentiation of human ESCs to retinal ganglion cells using a CRISPR engineered reporter cell line."],"pmcid":["PMC4643248"],"funding_grant_id":["R01EY023754","5P30EY001765","R01 EY023754","5T32EY007143","T32 EY007143","1R01EY02268001","P30 EY001765","T32-90040730"],"pubmed_authors":["Mao HQ","Marsh-Armstrong N","Davis CH","Krick K","Diamond JS","Sluch VM","Berlinicke CA","Zack DJ","Ranganathan V","Kerr JM","Martin R"],"additional_accession":[]},"is_claimable":false,"name":"Differentiation of human ESCs to retinal ganglion cells using a CRISPR engineered reporter cell line.","description":"Retinal ganglion cell (RGC) injury and cell death from glaucoma and other forms of optic nerve disease is a major cause of irreversible vision loss and blindness. Human pluripotent stem cell (hPSC)-derived RGCs could provide a source of cells for the development of novel therapeutic molecules as well as for potential cell-based therapies. In addition, such cells could provide insights into human RGC development, gene regulation, and neuronal biology. Here, we report a simple, adherent cell culture protocol for differentiation of hPSCs to RGCs using a CRISPR-engineered RGC fluorescent reporter stem cell line. Fluorescence-activated cell sorting of the differentiated cultures yields a highly purified population of cells that express a range of RGC-enriched markers and exhibit morphological and physiological properties typical of RGCs. Additionally, we demonstrate that aligned nanofiber matrices can be used to guide the axonal outgrowth of hPSC-derived RGCs for in vitro optic nerve-like modeling. Lastly, using this protocol we identified forskolin as a potent promoter of RGC differentiation.","dates":{"release":"2015-01-01T00:00:00Z","publication":"2015 Nov","modification":"2025-04-04T13:25:45.773Z","creation":"2019-03-27T02:01:52Z"},"accession":"S-EPMC4643248","cross_references":{"pubmed":["26563826"],"doi":["10.1038/srep16595"]}}