Skip to main content
SpringerLink
Log in
Book cover

Corneal Regeneration pp 425–436Cite as

Corneal Endothelium: Isolation and Cultivation Methods

  • Chapter
  • First Online:

Part of the book series: Essentials in Ophthalmology ((ESSENTIALS))

Abstract

The corneal endothelium maintains corneal transparency, and endothelial cell function depends on cell density and quality – with physiologic decline noted from childhood to adulthood. When this process is accelerated due to injuries or endothelial pathologies, irreversible corneal edema occurs. Traditionally transplantation has been the recourse for replacement of diseased corneal tissue; however this is limited by availability of cadaveric corneal tissue donor sources. There is considerable interest in developing suitable alternatives for donor endothelial graft material.

Recent advances have made possible the isolation and culture of suitable and viable human endothelial cells (HCEC) from various sources – whether from primary HCECs, established cell lines, or stem cells. Synthetic, biosynthetic, and organic substrates are also being investigated as carriers for the cultured cells. Current research is aimed at performing the culture process under xeno-free conditions. In conclusion, great progress has been made due to advances in knowledge of the HCEC cell cycle and the development of techniques for cell culture. This makes the application of tissue engineering for patients with corneal endothelial disease a distinct possibility in the near future.

This is a preview of subscription content, log in via an institution.

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD   199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

References

  1. Edelhauser HF. The balance between corneal transparency and edema the proctor lecture. Invest Opthalmol Vis Sci. 2006;47(5):1755.

    Article  Google Scholar 

  2. Srinivas SP. Dynamic regulation of barrier integrity of the corneal endothelium. Optom Vis Sci. 2010;87(4):1.

    Article  Google Scholar 

  3. Joyce NC. Proliferative capacity of corneal endothelial cells. Exp Eye Res. 2012;95(1):16–23.

    Article  CAS  PubMed  Google Scholar 

  4. Cui Y-B, Wu J. Research progress on the negative factors of corneal endothelial cells proliferation. Int J Ophthalmol. 2012;5(5):614–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Yee RW, Geroski DH, Matsuda M, Champeau EJ, Meyer LA, Edelhauser HF. Correlation of corneal endothelial pump site density, barrier function, and morphology in wound repair. Invest Ophthalmol Vis Sci. 1985;26(9):1191–201.

    CAS  PubMed  Google Scholar 

  6. Yee RW, Matsuda M, Schultz RO, Edelhauser HF. Changes in the normal corneal endothelial cellular pattern as a function of age. Curr Eye Res. 1985;4(6):671–8.

    Article  CAS  PubMed  Google Scholar 

  7. Wong KH, Kam KW, Chen LJ, Young AL. Corneal blindness and current major treatment concern-graft scarcity. Int J Ophthalmol. 2017;10(7):1154–62.

    PubMed  PubMed Central  Google Scholar 

  8. Pascolini D, Mariotti SP. Global estimates of visual impairment: 2010. Br J Ophthalmol. 2012;96(5):614–8.

    Article  PubMed  Google Scholar 

  9. Gain P, Jullienne R, He Z, Aldossary M, Acquart S, Cognasse F, et al. Global survey of corneal transplantation and eye banking. JAMA Ophthalmol. 2016;134(2):167.

    Article  PubMed  Google Scholar 

  10. Güell JL, El Husseiny MA, Manero F, Gris O, Elies D. Historical review and update of surgical treatment for corneal endothelial diseases. Ophthalmol Therapy. 2014;3(1–2):1–15.

    Article  Google Scholar 

  11. Lamm V, Hara H, Mammen A, Dhaliwal D, Cooper DKC. Corneal blindness and xenotransplantation. Xenotransplantation. 2014;21(2):99–114.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Borderie VM, Boëlle P-Y, Touzeau O, Allouch C, Boutboul S, Laroche L. Predicted long-term outcome of corneal transplantation. Ophthalmology. 2009;116(12):2354–60.

    Article  PubMed  Google Scholar 

  13. Vajpayee RB, Sharma N, Jhanji V, Titiyal JS, Tandon R. One donor cornea for 3 recipients: a new concept for corneal transplantation surgery. Arch Ophthalmol (Chicago Ill 1960). 2007;125(4):552–4.

    Article  Google Scholar 

  14. Okumura N, Koizumi N, Ueno M, Sakamoto Y, Takahashi H, Tsuchiya H, et al. ROCK inhibitor converts corneal endothelial cells into a phenotype capable of regenerating in vivo endothelial tissue. Am J Pathol. 2012;181(1):268–77.

    Article  CAS  PubMed  Google Scholar 

  15. Koizumi N, Sakamoto Y, Okumura N, Tsuchiya H, Torii R, Cooper LJ, et al. Cultivated corneal endothelial transplantation in a primate: possible future clinical application in corneal endothelial regenerative medicine. Cornea. 2008;27 Suppl 1:S48–55.

    Article  PubMed  Google Scholar 

  16. Okumura N, Koizumi N, Kay EP, Ueno M, Sakamoto Y, Nakamura S, et al. The ROCK inhibitor eye drop accelerates corneal endothelium wound healing. Invest Ophthalmol Vis Sci. 2013;54(4):2493–502.

    Article  CAS  PubMed  Google Scholar 

  17. Okumura N, Kinoshita S, Koizumi N. Application of rho kinase inhibitors for the treatment of corneal endothelial diseases. J Ophthalmol. 2017;2017:1–8.

    Google Scholar 

  18. Pipparelli A, Arsenijevic Y, Thuret G, Gain P, Nicolas M, Majo F. ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. Connon CJ, editor. PLoS One. 2013;8(4):e62095.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Proulx S, Brunette I. Methods being developed for preparation, delivery and transplantation of a tissue-engineered corneal endothelium. Exp Eye Res. 2012;95(1):68–75.

    Article  CAS  PubMed  Google Scholar 

  20. De Miguel MP, Alio JL, Arnalich-Montiel F, Fuentes-Julian S, de Benito-Llopis L, Amparo F, et al. Cornea and ocular surface treatment. Curr Stem Cell Res Ther. 2010;5(2):195–204.

    Article  PubMed  Google Scholar 

  21. Mannagh J, Irving AR. Human corneal endothelium: growth in tissue cultures. Arch Ophthalmol (Chicago Ill 1960). 1965;74(6):847–9.

    Article  CAS  Google Scholar 

  22. Zavala J, López Jaime GR, Rodríguez Barrientos CA, Valdez-Garcia J. Corneal endothelium: developmental strategies for regeneration. Eye. 2013;27(5):579–88.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Zhu C, Joyce NC. Proliferative Response of Corneal Endothelial Cells from Young and Older Donors. Invest Opthalmol Vis Sci. 2004;45(6):1743.

    Article  Google Scholar 

  24. Senoo T, Joyce NC. Cell cycle kinetics in corneal endothelium from old and young donors. Invest Ophthalmol Vis Sci. 2000;41(3):660–7.

    CAS  PubMed  Google Scholar 

  25. Joyce NC. Cell cycle status in human corneal endothelium. Exp Eye Res. 2005;81(6):629–38.

    Article  CAS  PubMed  Google Scholar 

  26. Miyata K, Drake J, Osakabe Y, Hosokawa Y, Hwang D, Soya K, et al. Effect of donor age on morphologic variation of cultured human corneal endothelial cells. Cornea. 2001;20(1):59–63.

    Article  CAS  PubMed  Google Scholar 

  27. Enomoto K, Mimura T, Harris DL, Joyce NC. Age differences in cyclin-dependent kinase inhibitor expression and rb hyperphosphorylation in human corneal endothelial cells. Invest Ophthalmol Vis Sci. 2006;47(10):4330–40.

    Article  PubMed  Google Scholar 

  28. Bednarz J, Rodokanaki-von Schrenck A, Engelmann K. Different characteristics of endothelial cells from central and peripheral human cornea in primary culture and after subculture. In Vitro Cell Dev Biol Anim. 1998;34(2):149–53.

    Article  CAS  PubMed  Google Scholar 

  29. Mimura T, Joyce NC. Replication competence and senescence in central and peripheral human corneal endothelium. Invest Ophthalmol Vis Sci. 2006;47(4):1387–96.

    Article  PubMed  Google Scholar 

  30. Raviola G. Schwalbe line’s cells: a new cell type in the trabecular meshwork of Macaca mulatta. Invest Ophthalmol Vis Sci. 1982;22(1):45–56.

    CAS  PubMed  Google Scholar 

  31. He Z, Campolmi N, Gain P, Ha Thi BM, Dumollard J-M, Duband S, et al. Revisited microanatomy of the corneal endothelial periphery: new evidence for continuous centripetal migration of endothelial cells in humans. Stem Cells. 2012;30(11):2523–34.

    Article  CAS  PubMed  Google Scholar 

  32. Whikehart DR, Parikh CH, Vaughn AV, Mishler K, Edelhauser HF. Evidence suggesting the existence of stem cells for the human corneal endothelium. Mol Vis. 2005;11:816–24.

    CAS  PubMed  Google Scholar 

  33. McGowan SL, Edelhauser HF, Pfister RR, Whikehart DR. Stem cell markers in the human posterior limbus and corneal endothelium of unwounded and wounded corneas. Mol Vis. 2007;13:1984–2000.

    Google Scholar 

  34. Choi SO, Jeon HS, Hyon JY, Oh Y-J, Wee WR, Chung T, et al. Recovery of corneal endothelial cells from periphery after injury. PLoS One. 2015;10(9):e0138076.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  35. Yokoo S, Yamagami S, Yanagi Y, Uchida S, Mimura T, Usui T, et al. Human corneal endothelial cell precursors isolated by sphere-forming assay. Invest Opthalmol Vis Sci. 2005;46(5):1626.

    Article  Google Scholar 

  36. Mimura T, Yamagami S, Yokoo S, Usui T, Amano S. Selective isolation of young cells from human corneal endothelium by the sphere-forming assay. Tissue Eng Part C Methods. 2010;16(4):803–12.

    Article  CAS  PubMed  Google Scholar 

  37. Yamagami S, Yokoo S, Mimura T, Takato T, Araie M, Amano S. Distribution of precursors in human corneal stromal cells and endothelial cells. Ophthalmology. 2007;114(3):433–9.

    Article  PubMed  Google Scholar 

  38. Bednarz J, Teifel M, Friedl P, Engelmann K. Immortalization of human corneal endothelial cells using electroporation protocol optimized for human corneal endothelial and human retinal pigment epithelial cells. Acta Ophthalmol Scand. 2000;78(2):130–6.

    Article  CAS  PubMed  Google Scholar 

  39. Valtink M, Gruschwitz R, Funk RHW, Engelmann K. Two clonal cell lines of immortalized human corneal endothelial cells show either differentiated or precursor cell characteristics. Cells Tissues Organs. 2008;187(4):286–94.

    Article  PubMed  Google Scholar 

  40. HCEC-B4G12 | Creative bioarray [Internet]. [cited 2018 Mar 13]. Available from: https://www.creative-bioarray.com/HCEC-B4G12-CSC-C3457-item-1468.htm.

  41. Kim H-J, Ryu Y-H, Ahn J-I, Park J-K, Kim J-C. Characterization of immortalized human corneal endothelial cell line using HPV 16 E6/E7 on lyophilized human amniotic membrane. Korean J Ophthalmol. 2006;20(1):47–54.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Schmedt T, Chen Y, Nguyen TT, Li S, Bonanno JA, Jurkunas UV. Telomerase immortalization of human corneal endothelial cells yields functional hexagonal monolayers. Lewin A, editor. PLoS One. 2012;7(12):e51427.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Yokoi T, Seko Y, Yokoi T, Makino H, Hatou S, Yamada M, et al. Establishment of functioning human corneal endothelial cell line with high growth potential. Kerkis I, editor. PLoS One. 2012;7(1):e29677.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Fan T, Zhao J, Ma X, Xu X, Zhao W, Xu B. Establishment of a continuous untransfected human corneal endothelial cell line and its biocompatibility to denuded amniotic membrane. Mol Vis. 2011;17:469–80.

    PubMed  PubMed Central  Google Scholar 

  45. Peh GSL, Beuerman RW, Colman A, Tan DT, Mehta JS. Human corneal endothelial cell expansion for corneal endothelium transplantation: an overview. Transplantation. 2011;91(8):811–9.

    Article  PubMed  Google Scholar 

  46. Engelmann K, Böhnke M, Friedl P. Isolation and long-term cultivation of human corneal endothelial cells. Invest Ophthalmol Vis Sci. 1988;29(11):1656–62.

    CAS  PubMed  Google Scholar 

  47. Peh GSL, Lee M-X, Wu F-Y, Toh K-P, Balehosur D, Mehta JS. Optimization of human corneal endothelial cells for culture: the removal of corneal stromal fibroblast contamination using magnetic cell separation. Int J Biomater. 2012;2012:601302.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  48. Parekh M, Baruzzo M, Favaro E, Borroni D, Ferrari S, Ponzin D, et al. Standardizing descemet membrane endothelial keratoplasty graft preparation method in the eye Bank — experience of 527 Descemet membrane endothelial keratoplasty tissues. Cornea. 2017;36(12):1458–1466.

    Article  PubMed  Google Scholar 

  49. Engelmann K, Bednarz J, Valtink M. Prospects for endothelial transplantation. Exp Eye Res. 2004;78(3):573–8.

    Article  CAS  PubMed  Google Scholar 

  50. Senoo T, Obara Y, Joyce NC. EDTA: a promoter of proliferation in human corneal endothelium. Invest Ophthalmol Vis Sci. 2000;41(10):2930–5.

    CAS  PubMed  Google Scholar 

  51. Li W, Sabater AL, Chen Y-T, Hayashida Y, Chen S-Y, He H, et al. A novel method of isolation, preservation, and expansion of human corneal endothelial cells. Invest Opthalmol Vis Sci. 2007;48(2):614.

    Article  Google Scholar 

  52. Peh GSL, Toh K-P, Wu F-Y, Tan DT, Mehta JS. Cultivation of human corneal endothelial cells isolated from paired donor corneas. Mohan RR, editor. PLoS One. 2011;6(12):e28310.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Gregory CD, Pound JD. Microenvironmental influences of apoptosis in vivo and in vitro. Apoptosis. 2010;15(9):1029–49.

    Article  CAS  PubMed  Google Scholar 

  54. Spinozzi D, Miron A, Bruinsma M, Lie JT, Dapena I, Oellerich S, et al. Improving the success rate of human corneal endothelial cell cultures from single donor corneas with stabilization medium. Cell Tissue Bank. 2017;19(1):9.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  55. Navaratnam J, Utheim T, Rajasekhar V, Shahdadfar A. Substrates for expansion of corneal endothelial cells towards bioengineering of human corneal endothelium. J Funct Biomater. 2015;6(3):917–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Chen KH, Azar D, Joyce NC. Transplantation of adult human corneal endothelium ex vivo: a morphologic study. Cornea. 2001;20(7):731–7.

    Article  CAS  PubMed  Google Scholar 

  57. Choi JS, Kim EY, Kim MJ, Giegengack M, Khan FA, Khang G, et al. In vitro evaluation of the interactions between human corneal endothelial cells and extracellular matrix proteins. Biomed Mater. 2013;8(1):14108.

    Article  CAS  Google Scholar 

  58. Engler C, Kelliher C, Speck CL, Jun AS. Assessment of attachment factors for primary cultured human corneal endothelial cells. Cornea. 2009;28(9):1050–4.

    Article  PubMed  Google Scholar 

  59. Choi JS, Williams JK, Greven M, Walter KA, Laber PW, Khang G, et al. Bioengineering endothelialized neo-corneas using donor-derived corneal endothelial cells and decellularized corneal stroma. Biomaterials. 2010;31(26):6738–45.

    Article  CAS  PubMed  Google Scholar 

  60. Yamaguchi M, Ebihara N, Shima N, Kimoto M, Funaki T, Yokoo S, et al. Adhesion, migration, and proliferation of cultured human corneal endothelial cells by laminin-5. Invest Ophthalmol Vis Sci. 2011;52(2):679–84.

    Article  CAS  PubMed  Google Scholar 

  61. Mimura T, Yamagami S, Yokoo S, Usui T, Tanaka K, Hattori S, et al. Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model. Invest Ophthalmol Vis Sci. 2004;45(9):2992–7.

    Article  PubMed  Google Scholar 

  62. Hitani K, Yokoo S, Honda N, Usui T, Yamagami S, Amano S. Transplantation of a sheet of human corneal endothelial cell in a rabbit model. Mol Vis. 2008;14:1–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  63. Numata R, Okumura N, Nakahara M, Ueno M, Kinoshita S, Kanematsu D, et al. Cultivation of corneal endothelial cells on a pericellular matrix prepared from human decidua-derived mesenchymal cells. Connon CJ, editor. PLoS One. 2014;9(2):e88169.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  64. Proulx S, Audet C, d’arc UJ, Deschambeault A, Carrier P, Giasson CJ, et al. Tissue engineering of feline corneal endothelium using a devitalized human cornea as carrier. Tissue Eng Part A. 2009;15(7):1709–18.

    Article  CAS  PubMed  Google Scholar 

  65. Honda N. Descemet stripping automated endothelial keratoplasty using cultured corneal endothelial cells in a rabbit model. Arch Ophthalmol. 2009;127(10):1321.

    Article  PubMed  Google Scholar 

  66. Bayyoud T, Thaler S, Hofmann J, Maurus C, Spitzer MS, Bartz-Schmidt K-U, et al. Decellularized bovine corneal posterior lamellae as carrier matrix for cultivated human corneal endothelial cells. Curr Eye Res. 2012;37(3):179–86.

    Article  CAS  PubMed  Google Scholar 

  67. Ishino Y, Sano Y, Nakamura T, Connon CJ, Rigby H, Fullwood NJ, et al. Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation. Invest Ophthalmol Vis Sci. 2004;45(3):800–6.

    Article  PubMed  Google Scholar 

  68. Wencan W, Mao Y, Wentao Y, Fan L, Jia Q, Qinmei W, et al. Using basement membrane of human amniotic membrane as a cell carrier for cultivated cat corneal endothelial cell transplantation. Curr Eye Res. 2007;32(3):199–215.

    Article  PubMed  Google Scholar 

  69. Niknejad H, Deihim T, Solati-Hashjin M, Peirovi H. The effects of preservation procedures on amniotic membrane’s ability to serve as a substrate for cultivation of endothelial cells. Cryobiology. 2011;63(3):145–51.

    Article  CAS  PubMed  Google Scholar 

  70. Yoeruek E, Saygili O, Spitzer MS, Tatar O, Bartz-Schmidt KU, Szurman P. Human anterior lens capsule as carrier matrix for cultivated human corneal endothelial cells. Cornea. 2009;28(4):416–20.

    Article  PubMed  Google Scholar 

  71. Kopsachilis N, Tsinopoulos I, Tourtas T, Kruse FE, Luessen UW. Descemet’s membrane substrate from human donor lens anterior capsule. Clin Exp Ophthalmol. 2012;40(2):187–94.

    Article  PubMed  Google Scholar 

  72. Van den Bogerd B, Dhubhghaill SN, Zakaria N. Characterizing Human Decellularized Crystalline Lens Capsules as a Scaffold for Corneal Endothelial Tissue Engineering. J Tissue Eng Regen Med. 2018:1–9.

    Google Scholar 

  73. Hsiue G-H, Lai J-Y, Chen K-H, Hsu W-M. A novel strategy for corneal endothelial reconstruction with a bioengineered cell sheet. Transplantation. J Tissue Eng Regen Med. 2018;12(4):e2020–8.

    Google Scholar 

  74. Lai J-Y, Chen K-H, Hsiue G-H. Tissue-engineered human corneal endothelial cell sheet transplantation in a rabbit model using functional biomaterials. Transplantation. 2007;84(10):1222–32.

    PubMed  Google Scholar 

  75. Yoeruek E, Bayyoud T, Maurus C, Hofmann J, Spitzer MS, Bartz-Schmidt K-U, et al. Decellularization of porcine corneas and repopulation with human corneal cells for tissue-engineered xenografts. Acta Ophthalmol. 2012;90(2):e125–31.

    Article  PubMed  Google Scholar 

  76. Joyce NC, Zhu CC. Human corneal endothelial cell proliferation: potential for use in regenerative medicine. Cornea. 2004;23(8 Suppl):S8–19.

    Article  PubMed  Google Scholar 

  77. Okumura N, Inoue R, Kakutani K, Nakahara M. Corneal endothelial cells have an absolute requirement for cysteine for survival. Cornea. 2017;36(8):988–94.

    Article  PubMed  Google Scholar 

  78. Peh GSL, Chng Z, Ang H-P, Cheng TYD, Adnan K, Seah X-Y, et al. Propagation of human corneal endothelial cells: a novel dual media approach. Cell Transplant. 2015;24(2):287–304.

    Article  PubMed  Google Scholar 

  79. Liu X, Tseng SC, Zhang M-C, Chen S-Y, Tighe S, Lu W-J, et al. LIF-JAK1-STAT3 signaling delays contact inhibition of human corneal endothelial cells. Cell Cycle. 2015;14(8):1197–206.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Engelmann K, Friedl P. Growth of human corneal endothelial cells in a serum-reduced medium. Cornea. 1995;14(1):62–70.

    Article  CAS  PubMed  Google Scholar 

  81. Blake DA, Yu H, Young DL, Caldwell DR. Matrix stimulates the proliferation of human corneal endothelial cells in culture. Invest Ophthalmol Vis Sci. 1997;38(6):1119–29.

    CAS  PubMed  Google Scholar 

  82. Jäckel T, Knels L, Valtink M, Funk RHW, Engelmann K. Serum-free corneal organ culture medium (SFM) but not conventional minimal essential organ culture medium (MEM) protects human corneal endothelial cells from apoptotic and necrotic cell death. Br J Ophthalmol. 2011;95(1):123–30.

    Article  PubMed  Google Scholar 

  83. Feizi S, Soheili Z-S, Bagheri A, Balagholi S, Mohammadian A, Rezaei-Kanavi M, et al. Effect of amniotic fluid on the in vitro culture of human corneal endothelial cells. Exp Eye Res. 2014;122:132–40.

    Article  CAS  PubMed  Google Scholar 

  84. Zhu Y-T, Li F, Han B, Tighe S, Zhang S, Chen S-Y, et al. Activation of RhoA-ROCK-BMP signaling reprograms adult human corneal endothelial cells. J Cell Biol. 2014;206(6):799–811.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Thieme D, Reuland L, Lindl T, Kruse F, Fuchsluger T. Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture. J Tissue Eng Regen Med. 2018;12(2):557–64.

    Article  CAS  PubMed  Google Scholar 

  86. Lee JG, Kay EP. FGF-2-mediated signal transduction during endothelial mesenchymal transformation in corneal endothelial cells. Exp Eye Res. 2006;83(6):1309–16.

    Article  CAS  PubMed  Google Scholar 

  87. Zhu Y-T, Chen H-C, Chen S-Y, Tseng SCG. Nuclear p120 catenin unlocks mitotic block of contact-inhibited human corneal endothelial monolayers without disrupting adherent junctions. J Cell Sci. 2012;125(Pt 15):3636–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  88. Zhu Y-T, Tighe S, Chen S-L, John T, Kao WY, Tseng SCG. Engineering of human corneal endothelial grafts. Curr Ophthalmol Rep. 2015;3(3):207–17.

    Article  PubMed  PubMed Central  Google Scholar 

  89. Lee JG, Ko MK, Kay EP. Endothelial mesenchymal transformation mediated by IL-1β-induced FGF-2 in corneal endothelial cells. Exp Eye Res. 2012;95(1):35–9.

    Article  CAS  PubMed  Google Scholar 

  90. Liu Y, Sun H, Hu M, Zhu M, Tighe S, Chen S, et al. Human corneal endothelial cells expanded in vitro are a powerful resource for tissue engineering. Int J Med Sci. 2017;14(2):128–35.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  91. Bartakova A, Alvarez-Delfin K, Weisman AD, Salero E, Raffa GA, Merkhofer RM, et al. Novel identity and functional markers for human corneal endothelial cells. Invest Opthalmol Vis Sci. 2016;57(6):2749.

    Article  CAS  Google Scholar 

  92. Frausto RF, Le DJ, Aldave AJ. Transcriptomic analysis of cultured corneal endothelial cells as a validation for their use in cell replacement therapy. Cell Transplant. 2016;25(6):1159–76.

    Article  PubMed  Google Scholar 

  93. Ding V, Chin A, Peh G, Mehta JS, Choo A. Generation of novel monoclonal antibodies for the enrichment and characterization of human corneal endothelial cells (hCENC) necessary for the treatment of corneal endothelial blindness. MAbs. 2014;6(6):1439–52.

    Article  PubMed  PubMed Central  Google Scholar 

  94. Dorfmueller S, Tan HC, Ngoh ZX, Toh KY, Peh G, Ang H-P, et al. Isolation of a recombinant antibody specific for a surface marker of the corneal endothelium by phage display. Sci Rep. 2016;6(1):21661.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  95. Koizumi N, Kinoshita S, Okumura N. The role of rho kinase inhibitors in corneal endothelial dysfunction. Curr Pharm Des. 2017;23(4):660–6.

    Article  CAS  PubMed  Google Scholar 

  96. Wang TJ, Chen MS, Chou ML, Lin HC, Seghatchian J, Burnouf T. Comparison of three human platelet lysates used as supplements for in vitro expansion of corneal endothelium cells. Transfus Apher Sci. 2017;56(5):769–73.

    Article  PubMed  Google Scholar 

  97. Peh GSL, Ang H-P, Lwin CN, Adnan K, George BL, Seah X-Y, et al. Regulatory compliant tissue-engineered human corneal endothelial grafts restore corneal function of rabbits with bullous keratopathy. Sci Rep. 2017;7(1):14149.

    Google Scholar 

Download references

Compliance with Ethical Requirements

David Mingo-Botín, Marie Joan Therese Dr. Balgos, and Francisco Arnalich-Montiel declare that they have no conflict of interest. No human or animal studies were carried out by the authors for this article.

Author information

Authors and Affiliations

  1. Cornea Unit, Department of Ophthalmology, Hospital Universitario Ramón y Cajal, Madrid, Spain

    David Mingo-Botín

  2. Research and Development Department, Cornea, Cataract and Refractive Surgery Department, Vissum Alicante, Alicante, Spain

    Marie Joan Therese D. Balgos

  3. Vissum Corporation, Madrid, Spain

    Francisco Arnalich-Montiel

Authors
  1. David Mingo-Botín
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marie Joan Therese D. Balgos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Francisco Arnalich-Montiel
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Miguel Hernández University , Alicante, Spain

    Jorge L. Alió

  2. Vissum-Instituto Oftalmologico de Alica, University Miguel Hernandez, Alicante, Alicante, Spain

    Jorge L. Alió del Barrio

  3. Vissum Corporation, Madrid, Spain

    Francisco Arnalich-Montiel

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Nature Switzerland AG

About this chapter

Check for updates. Verify currency and authenticity via CrossMark

Cite this chapter

Mingo-Botín, D., Balgos, M.J.T.D., Arnalich-Montiel, F. (2019). Corneal Endothelium: Isolation and Cultivation Methods. In: Alió, J., Alió del Barrio, J., Arnalich-Montiel, F. (eds) Corneal Regeneration . Essentials in Ophthalmology. Springer, Cham. https://doi.org/10.1007/978-3-030-01304-2_28

Download citation

  • DOI: https://doi.org/10.1007/978-3-030-01304-2_28

  • Published:

  • Publisher Name: Springer, Cham

  • Print ISBN: 978-3-030-01303-5

  • Online ISBN: 978-3-030-01304-2

  • eBook Packages: MedicineMedicine (R0)

Publish with us

Policies and ethics

Search

Navigation