• Valentin M Sluch, Chung-ha O Davis, Vinod Ranganathan, Justin M Kerr, Kellin Krick, Russ Martin, Cynthia A Berlinicke, Nicholas Marsh-Armstrong, Jeffrey S Diamond, Hai-Quan Mao, and Donald J Zack.
• Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine Baltimore, MD 21287.
• Sci Rep. 2015 Nov 13; 5: 16595.

AbstractRetinal 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.

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