The corneal endothelium is believed to be developmentally originated from the periocular mesenchyme via the neural crest. Human corneal endothelial progenitor cells (HCEPs) have been investigated because of their potential availability for the tissue regenerative medicine. However, the existence and the properties of HCEPs have not been elucidated yet. We first established a novel serum-free culture system for HCEPs. The HCEPs highly expressed p75 neurotrophin receptor, SOX9, and FOXC2, and partially retained the properties of neural crest and periocular mesenchyme. Further, we demonstrated that HCEPs had a high proliferative potency, and the differentiated HCEP sheets had corneal endothelial function by using the Ussing chamber system and transplantation to the rabbit cornea. These findings suggest that the HCEPs can be selectively expanded from the corneal endothelium using a specific culture system and will provide cell sheets for corneal regenerative medicine.
Get full access to this article
View all access options for this article.
References
1.
MauriceDM. (1972). The location of the fluid pump in the cornea. J Physiol, 221:43–54.
2.
LorenzettiDW, UotilaMH, ParikhN and KaufmanHE. (1967). Central cornea guttata. Incidence in the general population. Am J Ophthalmol, 64:1155–1158.
3.
SumideT, NishidaK, YamatoM, IdeT, HayashidaY, WatanabeK, YangJ, KohnoC, KikuchiA, et al. (2006). Functional human corneal endothelial cell sheets harvested from temperature-responsive culture surfaces. FASEB J, 20:392–394.
4.
WatanabeR, HayashiR, KimuraY, TanakaY, KageyamaT, HaraS, TabataY and NishidaK. (2011). A novel gelatin hydrogel carrier sheet for corneal endothelial transplantation. Tissue Eng Part A, 17:2213–2219.
5.
IdeT, NishidaK, YamatoM, SumideT, UtsumiM, NozakiT, KikuchiA, OkanoT and TanoY. (2006). Structural characterization of bioengineered human corneal endothelial cell sheets fabricated on temperature-responsive culture dishes. Biomaterials, 27:607–614.
6.
MimuraT, YamagamiS, YokooS, UsuiT, TanakaK, HattoriS, IrieS, MiyataK, AraieM and AmanoS. (2004). Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model. Invest Ophthalmol Vis Sci, 45:2992–2997.
7.
IshinoY, SanoY, NakamuraT, ConnonCJ, RigbyH, FullwoodNJ and KinoshitaS. (2004). Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation. Invest Ophthalmol Vis Sci, 45:800–806.
8.
ChoiJS, WilliamsJK, GrevenM, WalterKA, LaberPW, KhangG and SokerS. (2010). Bioengineering endothelialized neo-corneas using donor-derived corneal endothelial cells and decellularized corneal stroma. Biomaterials, 31:6738–6745.
9.
PehGS, BeuermanRW, ColmanA, TanDT and MehtaJS. (2011). Human corneal endothelial cell expansion for corneal endothelium transplantation: an overview. Transplantation, 91:811–819.
10.
MimuraT, YamagamiS and AmanoS. (2013). Corneal endothelial regeneration and tissue engineering. Prog Retin Eye Res, 35:1–17.
11.
YokooS, YamagamiS, YanagiY, UchidaS, MimuraT, UsuiT and AmanoS. (2005). Human corneal endothelial cell precursors isolated by sphere-forming assay. Invest Ophthalmol Vis Sci, 46:1626–1631.
12.
YamagamiS, YokooS, MimuraT, TakatoT, AraieM and AmanoS. (2007). Distribution of precursors in human corneal stromal cells and endothelial cells. Ophthalmology, 114:433–439.
13.
WhikehartDR, ParikhCH, VaughnAV, MishlerK and EdelhauserHF. (2005). Evidence suggesting the existence of stem cells for the human corneal endothelium. Mol Vis, 11:816–824.
14.
McGowanSL, EdelhauserHF, PfisterRR and WhikehartDR. (2007). Stem cell markers in the human posterior limbus and corneal endothelium of unwounded and wounded corneas. Mol Vis, 13:1984–2000.
15.
HeZ, CampolmiN, GainP, Ha ThiBM, DumollardJM, DubandS, Peoc'hM, PiselliS, GarraudO and ThuretG. (2012). Revisited microanatomy of the corneal endothelial periphery: new evidence for continuous centripetal migration of endothelial cells in humans. Stem Cells, 30:2523–2534.
16.
JohnstonMC, NodenDM, HazeltonRD, CoulombreJL and CoulombreAJ. (1979). Origins of avian ocular and periocular tissues. Exp Eye Res, 29:27–43.
17.
BahnCF, FallsHF, VarleyGA, MeyerRF, EdelhauserHF and BourneWM. (1984). Classification of corneal endothelial disorders based on neural crest origin. Ophthalmology, 91:558–563.
18.
HayashiK, SueishiK, TanakaK and InomataH. (1986). Immunohistochemical evidence of the origin of human corneal endothelial cells and keratocytes. Graefes Arch Clin Exp Ophthalmol, 224:452–456.
19.
YokoiT, SekoY, MakinoH, HatouS, YamadaM, KiyonoT, UmezawaA, NishinaH and AzumaN. (2012). Establishment of functioning human corneal endothelial cell line with high growth potential. PLoS One, 7:e29677
20.
HatouS, YoshidaS, HigaK, MiyashitaH, InagakiE, OkanoH, TsubotaK and ShimmuraS. (2013). Functional corneal endothelium derived from corneal stroma stem cells of neural crest origin by retinoic acid and Wnt/beta-catenin signaling. Stem Cells Dev, 22:828–839.
21.
GagePJ, RhoadesW, PruckaSK and HjaltT. (2005). Fate maps of neural crest and mesoderm in the mammalian eye. Invest Ophthalmol Vis Sci, 46:4200–4208.
22.
Hirata-TominagaK, NakamuraT, OkumuraN, KawasakiS, KayEP, BarrandonY, KoizumiN and KinoshitaS. (2013). Corneal endothelial cell fate is maintained by LGR5 via the regulation of Hedgehog and Wnt pathway. Stem Cells, 31:1396–1407.
23.
MorrisonSJ, WhitePM, ZockC and AndersonDJ. (1999). Prospective identification, isolation by flow cytometry, and in vivo self-renewal of multipotent mammalian neural crest stem cells. Cell, 96:737–749.
24.
CheungM and BriscoeJ. (2003). Neural crest development is regulated by the transcription factor Sox9. Development, 130:5681–5693.
25.
Sieber-BlumM, GrimM, HuYF and SzederV. (2004). Pluripotent neural crest stem cells in the adult hair follicle. Dev Dyn, 231:258–269.
26.
LothianC and LendahlU. (1997). An evolutionarily conserved region in the second intron of the human nestin gene directs gene expression to CNS progenitor cells and to early neural crest cells. Eur J Neurosci, 9:452–462.
27.
BattulaVL, BareissPM, TremlS, ConradS, AlbertI, HojakS, AbeleH, ScheweB, JustL, SkutellaT and BuhringHJ. (2007). Human placenta and bone marrow derived MSC cultured in serum-free, b-FGF-containing medium express cell surface frizzled-9 and SSEA-4 and give rise to multilineage differentiation. Differentiation, 75:279–291.
28.
SchuldinerM, YanukaO, Itskovitz-EldorJ, MeltonDA and BenvenistyN. (2000). Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A, 97:11307–11312.
29.
LiW, SabaterAL, ChenYT, HayashidaY, ChenSY, HeH and TsengSC. (2007). A novel method of isolation, preservation, and expansion of human corneal endothelial cells. Invest Ophthalmol Vis Sci, 48:614–620.
30.
BlakeDA, YuH, YoungDL and CaldwellDR. (1997). Matrix stimulates the proliferation of human corneal endothelial cells in culture. Invest Ophthalmol Vis Sci, 38:1119–1129.
31.
ZhuC and JoyceNC. (2004). Proliferative response of corneal endothelial cells from young and older donors. Invest Ophthalmol Vis Sci, 45:1743–1751.
32.
RodinS, DomogatskayaA, StromS, HanssonEM, ChienKR, InzunzaJ, HovattaO and TryggvasonK. (2010). Long-term self-renewal of human pluripotent stem cells on human recombinant laminin-511. Nat Biotechnol, 28:611–615.
33.
KabosovaA, AzarDT, BannikovGA, CampbellKP, DurbeejM, GhohestaniRF, JonesJC, KenneyMC, KochM, et al. (2007). Compositional differences between infant and adult human corneal basement membranes. Invest Ophthalmol Vis Sci, 48:4989–4999.
34.
SenooT, ObaraY and JoyceNC. (2000). EDTA: a promoter of proliferation in human corneal endothelium. Invest Ophthalmol Vis Sci, 41:2930–2935.
35.
LuX, ChenD, LiuZ, LiC, LiuY, ZhouJ, WanP, MouYG and WangZ. (2010). Enhanced survival in vitro of human corneal endothelial cells using mouse embryonic stem cell conditioned medium. Mol Vis, 16:611–622.
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.
