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Publication
Intrinsic Morphologic and Physiologic Development of Human Derived Retinal
Ganglion Cells In Vitro.
Authors Risner ML, Pasini S, Chamling X, McGrady NR, Goldberg JL, Zack DJ, Calkins DJ
Submitted By Submitted Externally on 12/3/2021
Status Published
Journal Translational vision science & technology
Year 2021
Date Published 8/1/2021
Volume : Pages 10 : 1
PubMed Reference 34383881
Abstract Human retinal ganglion cells (hRGC) derived from human pluripotent stem cells
are promising candidates to model, protect, and replace degenerating RGCs. Here,
we examined intrinsic morphologic and physiologic development of hRGCs., We used
CRISPR-Cas9 to selectively express tdTomato under the RGC-specific promoter,
BRN3B. Human pluripotent stem cells were chemically differentiated into hRGCs
and cultured up to 7 weeks. We measured soma area, neurite complexity, synaptic
protein, axon-related messenger RNA and protein, and voltage-dependent
responses., Soma area, neurite complexity, and postsynaptic density protein 95
increased over time. Soma area and neurite complexity increased proportionally
week to week, and this relationship was dynamic, strengthening between 2 and 3
weeks and diminishing by 4 weeks. Postsynaptic density 95 localization was
dependent on culture duration. After 1 to 2 weeks, postsynaptic density 95
localized within somas but redistributed along neurites after 3 to 4 weeks. Axon
initial segment scaffolding protein, Ankyrin G, expression also increased over
time, and by 7 weeks, Ankyrin G often localized within putative axons.
Voltage-gated inward currents progressively developed, but outward currents
matured by 4 weeks. Current-induced spike generation increased over time but
limited by depolarization block., Human RGCs develop up to 7 weeks after
culture. Thus, the state of hRGC maturation should be accounted for in designing
models and treatments for optic neuropathies., We characterized hRGC morphologic
and physiologic development towards identifying key time points when hRGCs
express mechanisms that may be harnessed to enhance the efficacy of
neuroprotective and cell replacement therapies.




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