title: Enriched retinal ganglion cells derived from human embryonic stem cells.
abstract: Optic neuropathies are characterised by a loss of retinal ganglion cells (RGCs) that lead to vision impairment. Development of cell therapy requires a better understanding of the signals that direct stem cells into RGCs. Human embryonic stem cells (hESCs) represent an unlimited cellular source for generation of human RGCs in vitro. In this study, we present a 45-day protocol that utilises magnetic activated cell sorting to generate enriched population of RGCs via stepwise retinal differentiation using hESCs. We performed an extensive characterization of these stem cell-derived RGCs by examining the gene and protein expressions of a panel of neural/RGC markers. Furthermore, whole transcriptome analysis demonstrated similarity of the hESC-derived RGCs to human adult RGCs. The enriched hESC-RGCs possess long axons, functional electrophysiological profiles and axonal transport of mitochondria, suggestive of maturity. In summary, this RGC differentiation protocol can generate an enriched population of functional RGCs from hESCs, allowing future studies on disease modeling of optic neuropathies and development of cell therapies.
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Katherine P
authors: KP
authors: Gill
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Sandy S C
authors: SS
authors: Hung
authors: National Institute of Aging, National Institutes for Health, USA.
authors: Alexei
authors: A
authors: Sharov
authors: Monash Micro Imaging, Monash University, Australia.
authors: Camden Y
authors: CY
authors: Lo
authors: Department of Otolaryngology &Department of Surgery, the University of Melbourne, and Royal Victorian Eye and Ear Hospital, Australia.
authors: Karina
authors: K
authors: Needham
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Grace E
authors: GE
authors: Lidgerwood
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Stacey
authors: S
authors: Jackson
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Duncan E
authors: DE
authors: Crombie
authors: Departments of Audiology and Speech Pathology &Ophthalmology, University of Melbourne, Australia.
authors: Bryony A
authors: BA
authors: Nayagam
authors: Wicking Dementia Research and Education Centre, University of Tasmania, Australia.
authors: Anthony L
authors: AL
authors: Cook
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.|School of Medicine, Menzies Institute for Medical Research, University of Tasmania, Australia.
authors: Alex W
authors: AW
authors: Hewitt
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Alice
authors: A
authors: Pébay
authors: Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital &Ophthalmology, Department of Surgery, the University of Melbourne, Australia.
authors: Raymond C B
authors: RC
authors: Wong
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D060526:Magnetic Fields; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers