Restricted accessResearch articleFirst published online 2019-2
Transplantation of Retinal Progenitor Cells from Optic Cup-Like Structures Differentiated from Human Embryonic Stem Cells In Vitro and In Vivo Generation of Retinal Ganglion-Like Cells
Human embryonic stem cells (hESCs) have the potential to differentiate along the retinal lineage. We have efficiently differentiated human pluripotent stem cells into optic cup-like structures by using a novel retinal differentiation medium (RDM). The purpose of this study was to determine whether the retinal progenitor cells (RPCs) derived from hESCs can integrate into the host retina and differentiate into retinal ganglion cells (RGCs) in vivo. In this study, hESCs (H9-GFP) were induced to differentiate into optic cup-like structures by using our novel differentiation system. The RPCs extracted from the optic cup-like structures were transplanted into the vitreous cavity of N-methyl-d-aspartic acid-treated mice. Sham-treated eyes received the same amount of RDM. The host retinas were analyzed by triple immunofluorescence on the fourth and fifth weeks after transplantation. The optic cup-like structures were efficiently differentiated from hESCs by using our novel differentiation system in vitro for 6–8 weeks. The RPCs extracted from the optic cup-like structures migrated and integrated into the ganglion cell layer (GCL) of the host retina. Furthermore, the remaining transplanted cells were spread over the GCL and had a complementary distribution with host residual RGCs in the GCL of the mouse retina. Surprisingly, some of the transplanted cells expressed the RGC-specific marker Brn3a. These findings demonstrated that the RPCs derived from hESCs could integrate into the host GCL and differentiate into retinal ganglion-like cells in vivo, suggesting that RPCs can be used as an ideal source in supplying countless RGC and embryonic stem cell-based replacement therapies may be a promising treatment to restore vision in patients with degenerative retinal diseases.
Get full access to this article
View all access options for this article.
References
1.
QuigleyHA and BromanAT. (2006). The number of people with glaucoma worldwide in 2010 and 2020. Brit J Ophthalmol, 90:262–267.
2.
SchmierJK, JonesML and HalpernMT. (2006). The burden of age-related macular degeneration. Pharmacoeconomics, 24:319–334.
3.
HoAC, HumayunMS, DornJD, da CruzL, DagnelieG, HandaJ, BaralePO, SahelJA, StangaPE, et al. (2015). Long-term results from an epiretinal prosthesis to restore sight to the blind. Ophthalmol, 122:1547–1554.
4.
WangY, XuK, ZhangH, ZhaoJ, ZhuX, WangY and WuR. (2014). Retinal ganglion cell death is triggered by paraptosis via reactive oxygen species production: a brief literature review presenting a novel hypothesis in glaucoma pathology. Mol Med Rep, 10:1179–1183.
KjartansdóttirKR, GabrielsenA, RedaA, SöderO, Bergström-TengzeliusR, AndersenCY, HovattaO, StukenborgJB and FedderJ. (2012). Differentiation of stem cells upon deprivation of exogenous FGF2: a general approach to study spontaneous differentiation of hESCs in vitro. Syst Biol Reprod Med, 58:330–338.
7.
NistorG, SeilerMJ, YanF, FergusonD and KeirsteadHS. (2010). Three-dimensional early retinal progenitor 3D tissue constructs derived from human embryonic stem cells. J Neurosci Meth, 190:63–70.
8.
NguyenHV, LiY and TsangSH. (2015). Patient-specific iPSC-derived RPE for modeling of retinal diseases. J Clin Med, 4:567–578.
9.
SluchVM, DavisCO, RanganathanV, KerrJM, KrickK, MartinR, BerlinickeCA, Marsh-ArmstrongN, DiamondJS, MaoHQ and ZackDJ. (2015). Differentiation of human ESCs to retinal ganglion cells using a CRISPR engineered reporter cell line. Sci Rep, 5:16595.
10.
Barnea-CramerAO, WangW, LuSJ, SinghMS, LuoC, HuoH, McClementsME, BarnardAR, MacLarenRE and LanzaR. (2016). Function of human pluripotent stem cell-derived photoreceptor progenitors in blind mice. Sci Rep, 6:29784.
11.
IdelsonM, AlperR, ObolenskyA, Ben-ShushanE, HemoI, Yachimovich-CohenN, KhanerH, SmithY, WiserO, et al. (2009). Directed differentiation of human embryonic stem cells into functional retinal pigment epithelium cells. Cell Stem Cell, 5:396–408.
12.
NakanoT, AndoS, TakataN, KawadaM, MugurumaK, SekiguchiK, SaitoK, YonemuraS, EirakuM and SasaiY. (2012). Self-formation of optic cups and storable stratified neural retina from human ESCs. Cell Stem Cell, 10:771–785.
13.
da CruzL, FynesK, GeorgiadisO, KerbyJ, LuoYH, AhmadoA, VernonA, DanielsJT, NommisteB, et al. (2018). Phase 1 clinical study of an embryonic stem cell-derived retinal pigment epithelium patch in age-related macular degeneration. Nat Biotechnol, 36:328–337.
14.
Lopez-OnievaL, MontesR, LamoldaM, RomeroT, AyllonV, LozanoML, VicenteV, RiveraJ, Ramos-MejíaV and RealPJ. (2016). Generation of induced pluripotent stem cells (iPSCs) from a Bernard–Soulier syndrome patient carrying a W71R mutation in the GPIX gene. Stem Cell Res, 16:692–695.
15.
SchwartzSD, RegilloCD, LamBL, EliottD, RosenfeldPJ, GregoriNZ, HubschmanJP, DavisJL, HeilwellG, et al. (2015). Human embryonic stem cell-derived retinal pigment epithelium in patients with age-related macular degeneration and Stargardt's macular dystrophy: follow-up of two open-label phase 1/2 studies. Lancet, 385:509–516.
16.
LambaDA, GustJ and RehTA. (2009). Transplantation of human embryonic stem cell-derived photoreceptors restores some visual function in Crx-deficient mice. Cell Stem Cell, 4:73–79.
17.
SchwartzSD, HubschmanJP, HeilwellG, Franco-CardenasV, PanCK, OstrickRM, MickunasE, GayR, KlimanskayaI and LanzaR. (2012). Embryonic stem cell trials for macular degeneration: a preliminary report. Lancet, 379:713–720.
18.
HazimRA, KarumbayaramS, JiangM, DimashkieA, LopesVS, LiD, BurgessBL, VijayarajP, Alva-OrnelasJA, et al. (2017). Differentiation of RPE cells from integration-free iPS cells and their cell biological characterization. Stem Cell Res Ther, 8:217.
19.
MandaiM, WatanabeA, KurimotoY, HiramiY, MorinagaC, DaimonT, FujiharaM, AkimaruH, SakaiN, et al. (2017). Autologous induced stem-cell-derived retinal cells for macular degeneration. N Engl J Med, 376:1038–1046.
20.
KamaoH, MandaiM, OhashiW, HiramiY, KurimotoY, KiryuJ and TakahashiM. (2017). Evaluation of the surgical device and procedure for extracellular matrix-scaffold-supported human iPSC-derived retinal pigment epithelium cell sheet transplantation. Invest Ophthalmol Vis Sci, 58:211–220.
21.
ZhongX, GutierrezC, XueT, HamptonC, VergaraMN, CaoLH, PetersA, ParkTS, ZambidisET, et al. (2014). Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs. Nat Commun, 5:4047.
22.
WangX, XiongK, LinC, LvL, ChenJ, XuC, WangS, GuD, ZhengH, et al. (2015). New medium used in the differentiation of human pluripotent stem cells to retinal cells is comparable to fetal human eye tissue. Biomaterials, 53:40–49.
23.
ZagozewskiJL, ZhangQ and EisenstatDD. (2014). Genetic regulation of vertebrate eye development. Clin Genet, 86:453–460.
24.
OhsawaR and KageyamaR. (2008). Regulation of retinal cell fate specification by multiple transcription factors. Brain Res, 1192:90–98.
25.
LeTT, WroblewskiE, PatelS, RiesenbergAN and BrownNL. (2006). Math5 is required for both early retinal neuron differentiation and cell cycle progression. Dev Biol, 295:764–778.
26.
BrownNL, PatelS, BrzezinskiJ and GlaserT. (2001). Math5 is required for retinal ganglion cell and optic nerve formation. Development, 128:2497–2508.
27.
FengL, XieZH, DingQ, XieXL, LibbyRT and GanL. (2010). MATH5 controls the acquisition of multiple retinal cell fates. Mol Brain, 3:36.
28.
WuZK, CaoL, ZhangXY, SongWT and XiaXB. (2016). Promotion on the differentiation of retinal Muller cells into retinal ganglion cells by Brn-3b. Int J Ophthalmol- Chi, 9:948–954.
29.
QiuF, JiangH and XiangM. (2008). A comprehensive negative regulatory program controlled by Brn3b to ensure ganglion cell specification from multipotential retinal precursors. J Neurosci, 28:3392–3403.
30.
ZhangJ, YangZ and WuSM. (2004). Immuocytochemical analysis of spatial organization of photoreceptors and amacrine and ganglion cells in the tiger salamander retina. Vis Neurosci, 21:157–166.
31.
WangS, GuD, ZhangP, ChenJ, LiY, XiaoH and ZhouG. (2018). Melanopsin-expressing retinal ganglion cells are relatively resistant to excitotoxicity induced by N-Methyl-D-aspartate. Neurosci Lett, 662:368–373.
32.
JagathaB, DivyaMS, SanalkumarR, IndulekhaCL, VidyanandS, DivyaTS, DasAV and JamesJ. (2009). In vitro differentiation of retinal ganglion-like cells from embryonic stem cell derived neural progenitors. Biochem Biophys Res Commun, 380:230–235.
33.
ChenM, ChenQ, SunX, ShenW, LiuB, ZhongX, LengY, LiC, ZhangW, et al. (2010). Generation of retinal ganglion-like cells from reprogrammed mouse fibroblasts. Invest Ophthalmol Vis Sci, 51:5970–5978.
34.
TeotiaP, ChopraDA, DravidSM, Van HookMJ, QiuF, MorrisonJ, RizzinoA and AhmadI. (2017). Generation of functional human retinal ganglion cells with target specificity from pluripotent stem cells by chemically defined recapitulation of developmental mechanism. Stem Cells, 35:572–585.
35.
FarahMH. (2006). Neurogenesis and cell death in the ganglion cell layer of vertebrate retina. Brain Res Rev, 52:264–274.
36.
PerryVH. (1981). Evidence for an amacrine cell system in the ganglion-cell layer of the rat retina. Neuroscience, 6:931–944.
37.
PangJJ and WuSM. (2011). Morphology and immunoreactivity of retrogradely double-labeled ganglion cells in the mouse retina. Invest Ophthalmol Vis Sci, 52:4886–4896.
38.
SiliprandiR, CanellaR, CarmignotoG, SchiavoN, ZanellatoA, ZanoniR and VantiniG. (1992). N-Methyl-D-aspartate-induced neurotoxicity in the adult rat retina. Vis Neurosci, 8:567–573.
39.
Agudo-BarriusoM, Villegas-PérezMP, de ImperialJM and Vidal-SanzM. (2013). Anatomical and functional damage in experimental glaucoma. Curr Opin Pharmacol, 13:5–11.
40.
HarrisB. (2008). Cellular determination in the xenopus retina is independent of lineage and birth date. Neuron, 60:395–396.
41.
WettsR and FraserSE. (1988). Multipotent precursors can give rise to all major cell-types of the frog retina. Science, 239:1142–1145.
42.
AokiH, HaraA, NiwaM, MotohashiT, SuzukiT and KunisadaT. (2008). Transplantation of cells from eye-like structures differentiated from embryonic stem cells in vitro and in vivo regeneration of retinal ganglion-like cells. Graefes Arch Clin Exp Ophthalmol, 246:255–265.
43.
NishidaA, TakahashiM, TaniharaH, NakanoI, TakahashiJB, MizoguchiA, IdeC and HondaY. (2000). Incorporation and differentiation of hippocampus-derived neural stem cells transplanted in injured adult rat retina. Invest Ophthalmol Vis Sci, 41:4268–4274.
44.
KurimotoY, ShibukiH, KanekoY, IchikawaM, KurokawaT, TakahashiM and YoshimuraN. (2001). Transplantation of adult rat hippocampus-derived neural stem cells into retina injured by transient ischemia. Neurosci Lett, 306:57–60.
45.
KielczewskiJL, PeaseME and QuigleyHA. (2005). The effect of experimental glaucoma and optic nerve transection on amacrine cells in the rat retina. Invest Ophthalmol Vis Sci, 46:3188–3196.
46.
ArnholdS, KleinH, SemkovaI, AddicksK and SchraermeyerU. (2004). Neurally selected embryonic stem cells induce tumor formation after long-term survival following engraftment into the subretinal space. Invest Ophthalmol Vis Sci, 45:4251–4255.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
