PMID- 34383881
OWN - NLM
STAT- MEDLINE
DCOM- 20211020
LR  - 20220803
IS  - 2164-2591 (Electronic)
IS  - 2164-2591 (Linking)
VI  - 10
IP  - 10
DP  - 2021 Aug 12
TI  - Intrinsic Morphologic and Physiologic Development of Human Derived Retinal 
      Ganglion Cells In Vitro.
PG  - 1
LID - 10.1167/tvst.10.10.1 [doi]
LID - 1
AB  - PURPOSE: 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. 
      METHODS: 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. RESULTS: 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. CONCLUSIONS: 
      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. TRANSLATIONAL RELEVANCE: 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.
FAU - Risner, Michael L
AU  - Risner ML
AD  - Vanderbilt Eye Institute, Department of Ophthalmology & Visual Sciences, 
      Vanderbilt University Medical Center, Nashville, TN, USA.
FAU - Pasini, Silvia
AU  - Pasini S
AD  - Vanderbilt Eye Institute, Department of Ophthalmology & Visual Sciences, 
      Vanderbilt University Medical Center, Nashville, TN, USA.
FAU - Chamling, Xitiz
AU  - Chamling X
AD  - Wilmer Eye Institute, Department of Ophthalmology, Johns Hopkins University 
      School of Medicine, Baltimore, MD, USA.
FAU - McGrady, Nolan R
AU  - McGrady NR
AD  - Vanderbilt Eye Institute, Department of Ophthalmology & Visual Sciences, 
      Vanderbilt University Medical Center, Nashville, TN, USA.
FAU - Goldberg, Jeffrey L
AU  - Goldberg JL
AD  - Byers Eye Institute, Department of Ophthalmology, Stanford University School of 
      Medicine, Palo Alto, CA, USA.
FAU - Zack, Donald J
AU  - Zack DJ
AD  - Wilmer Eye Institute, Department of Ophthalmology, Johns Hopkins University 
      School of Medicine, Baltimore, MD, USA.
FAU - Calkins, David J
AU  - Calkins DJ
AD  - Vanderbilt Eye Institute, Department of Ophthalmology & Visual Sciences, 
      Vanderbilt University Medical Center, Nashville, TN, USA.
LA  - eng
GR  - P30 EY026877/EY/NEI NIH HHS/United States
GR  - P30 DK058404/DK/NIDDK NIH HHS/United States
GR  - P30 EY001765/EY/NEI NIH HHS/United States
GR  - P30 EY008126/EY/NEI NIH HHS/United States
GR  - P30 CA068485/CA/NCI NIH HHS/United States
GR  - R01 EY024997/EY/NEI NIH HHS/United States
GR  - U24 EY029903/EY/NEI NIH HHS/United States
GR  - P30 DK020593/DK/NIDDK NIH HHS/United States
GR  - U24 DK059637/DK/NIDDK NIH HHS/United States
GR  - R01 EY017427/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Transl Vis Sci Technol
JT  - Translational vision science & technology
JID - 101595919
SB  - IM
MH  - Axons
MH  - Cell Differentiation
MH  - Humans
MH  - Neurites
MH  - *Pluripotent Stem Cells
MH  - *Retinal Ganglion Cells
PMC - PMC8362626
COIS- Disclosure: M.L. Risner, None; S. Pasini, None; X. Chamling, None; N.R. McGrady, 
      None; J.L. Goldberg, None; D.J. Zack, None; D.J. Calkins, None
EDAT- 2021/08/13 06:00
MHDA- 2021/10/21 06:00
PMCR- 2021/08/12
CRDT- 2021/08/12 17:30
PHST- 2021/08/12 17:30 [entrez]
PHST- 2021/08/13 06:00 [pubmed]
PHST- 2021/10/21 06:00 [medline]
PHST- 2021/08/12 00:00 [pmc-release]
AID - 2776589 [pii]
AID - TVST-21-3489 [pii]
AID - 10.1167/tvst.10.10.1 [doi]
PST - ppublish
SO  - Transl Vis Sci Technol. 2021 Aug 12;10(10):1. doi: 10.1167/tvst.10.10.1.