PMID- 26283078
OWN - NLM
STAT- MEDLINE
DCOM- 20160909
LR  - 20181113
IS  - 1557-8534 (Electronic)
IS  - 1547-3287 (Print)
IS  - 1547-3287 (Linking)
VI  - 24
IP  - 23
DP  - 2015 Dec 1
TI  - Characterization of Three-Dimensional Retinal Tissue Derived from Human Embryonic 
      Stem Cells in Adherent Monolayer Cultures.
PG  - 2778-95
LID - 10.1089/scd.2015.0144 [doi]
AB  - Stem cell-based therapy of retinal degenerative conditions is a promising 
      modality to treat blindness, but requires new strategies to improve the number of 
      functionally integrating cells. Grafting semidifferentiated retinal tissue rather 
      than progenitors allows preservation of tissue structure and connectivity in 
      retinal grafts, mandatory for vision restoration. Using human embryonic stem 
      cells (hESCs), we derived retinal tissue growing in adherent conditions 
      consisting of conjoined neural retina and retinal pigment epithelial (RPE) cells 
      and evaluated cell fate determination and maturation in this tissue. We found 
      that deriving such tissue in adherent conditions robustly induces all eye field 
      genes (RX, PAX6, LHX2, SIX3, SIX6) and produces four layers of pure populations 
      of retinal cells: RPE (expressing NHERF1, EZRIN, RPE65, DCT, TYR, TYRP, MITF, 
      PMEL), early photoreceptors (PRs) (coexpressing CRX and RCVRN), inner nuclear 
      layer neurons (expressing CALB2), and retinal ganglion cells [RGCs, expressing 
      BRN3B and Neurofilament (NF) 200]. Furthermore, we found that retinal progenitors 
      divide at the apical side of the hESC-derived retinal tissue (next to the RPE 
      layer) and then migrate toward the basal side, similar to that found during 
      embryonic retinogenesis. We detected synaptogenesis in hESC-derived retinal 
      tissue, and found neurons containing many synaptophysin-positive boutons within 
      the RGC and PR layers. We also observed long NF200-positive axons projected by 
      RGCs toward the apical side. Whole-cell recordings demonstrated that putative 
      amacrine and/or ganglion cells exhibited electrophysiological responses 
      reminiscent of those in normal retinal neurons. These responses included 
      voltage-gated Na(+) and K(+) currents, depolarization-induced spiking, and 
      responses to neurotransmitter receptor agonists. Differentiation in adherent 
      conditions allows generation of long and flexible pieces of 3D retinal tissue 
      suitable for isolating transplantable slices of tissue for retinal replacement 
      therapies.
FAU - Singh, Ratnesh K
AU  - Singh RK
AD  - 1 Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, 
      University of Pittsburgh Medical Center , Pittsburgh, Pennsylvania.
FAU - Mallela, Ramya K
AU  - Mallela RK
AD  - 1 Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, 
      University of Pittsburgh Medical Center , Pittsburgh, Pennsylvania.
FAU - Cornuet, Pamela K
AU  - Cornuet PK
AD  - 1 Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, 
      University of Pittsburgh Medical Center , Pittsburgh, Pennsylvania.
FAU - Reifler, Aaron N
AU  - Reifler AN
AD  - 2 Department of Ophthalmology and Visual Sciences, University of Michigan , Ann 
      Arbor, Michigan.
FAU - Chervenak, Andrew P
AU  - Chervenak AP
AD  - 2 Department of Ophthalmology and Visual Sciences, University of Michigan , Ann 
      Arbor, Michigan.
FAU - West, Michael D
AU  - West MD
AD  - 3 BioTime, Inc. , Alameda, California.
FAU - Wong, Kwoon Y
AU  - Wong KY
AD  - 2 Department of Ophthalmology and Visual Sciences, University of Michigan , Ann 
      Arbor, Michigan.
FAU - Nasonkin, Igor O
AU  - Nasonkin IO
AD  - 1 Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, 
      University of Pittsburgh Medical Center , Pittsburgh, Pennsylvania.
LA  - eng
GR  - P30 EY007003/EY/NEI NIH HHS/United States
GR  - P30 EY008098/EY/NEI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20150910
PL  - United States
TA  - Stem Cells Dev
JT  - Stem cells and development
JID - 101197107
RN  - 0 (Synaptophysin)
RN  - 9NEZ333N27 (Sodium)
RN  - RWP5GA015D (Potassium)
SB  - IM
MH  - Action Potentials
MH  - Cells, Cultured
MH  - Embryonic Stem Cells/*cytology/metabolism
MH  - Humans
MH  - Neurogenesis
MH  - Potassium/metabolism
MH  - Retinal Neurons/*cytology/metabolism
MH  - Retinal Pigment Epithelium/*cytology/metabolism
MH  - Sodium/metabolism
MH  - Synapses/metabolism/physiology
MH  - Synaptophysin/genetics/metabolism
MH  - *Tissue Engineering
PMC - PMC4653822
EDAT- 2015/08/19 06:00
MHDA- 2016/09/10 06:00
PMCR- 2016/12/01
CRDT- 2015/08/19 06:00
PHST- 2015/08/19 06:00 [entrez]
PHST- 2015/08/19 06:00 [pubmed]
PHST- 2016/09/10 06:00 [medline]
PHST- 2016/12/01 00:00 [pmc-release]
AID - 10.1089/scd.2015.0144 [pii]
AID - 10.1089/scd.2015.0144 [doi]
PST - ppublish
SO  - Stem Cells Dev. 2015 Dec 1;24(23):2778-95. doi: 10.1089/scd.2015.0144. Epub 2015 
      Sep 10.