Amniotic membrane maintains the phenotype of rabbit retinal pigment epithelial cells in culture

Boris V. Stanzel, Edgar M. Espana, Martin Grueterich, Tetsuya Kawakita, Jean Marie Parel, Scheffer C G Tseng, Susanne Binder

研究成果: Article

39 引用 (Scopus)

抄録

The success of surgical removal of choroidal neovascularisation followed by transplantation of autologous retinal pigment epithelial cells (RPE) for age-related macular degeneration (ARMD) may be limited by damage in Bruch's membrane. We investigated whether amniotic membrane (AM) might be used as an alternative basement membrane-containing matrix to support RPE growth and differentiation. Primary RPE plastic cultures were established from freshly enucleated Dutch belted rabbit eyes in DMEM/F12 containing 0·1 mm Ca ++ and 10% dialysed FBS. Upon subconfluence, cells were subcultured at 5000-9000 cells cm-2 in the above-mentioned culture medium on intact AM (iAM), epithelially denuded AM (dAM) or plastic. After confluence, the Ca++ concentration in the medium was increased to 1·8 mm for 4 weeks. Growth and morphology were monitored by phase contrast microscopy, and the phenotype by immunostaining with antibodies against cytokeratin 18, tight junction protein ZO-1, and RPE65 protein, and by transepithelial resistance (TER) measurement. Immunostaining to cytokeratin 18 confirmed the epithelial origin of isolated cells in both primary culture and subcultures. Compared to plastic cultures, RPE increased pigmentation within 24 hr after seeding on AM, with iAM being more pronounced than dAM. RPE adopted a hexagonal epithelial phenotype with more organised pigmentation, strong expression of ZO-1 and RPE65, and a significantly higher TER 4 weeks after Ca++ switch on dAM. Our results indicate that AM may be used as a basement membrane-containing matrix to maintain RPE phenotype in vitro, and may facilitate subsequent transplantation to treat ARMD.

元の言語English
ページ(範囲)103-112
ページ数10
ジャーナルExperimental Eye Research
80
発行部数1
DOI
出版物ステータスPublished - 2005 1
外部発表Yes

Fingerprint

Amnion
Retinal Pigments
Cell Culture Techniques
Epithelial Cells
Rabbits
Phenotype
Plastics
Keratin-18
Pigmentation
Macular Degeneration
Basement Membrane
Zonula Occludens-1 Protein
Bruch Membrane
Phase-Contrast Microscopy
Choroidal Neovascularization
Autologous Transplantation
Growth
Culture Media
Cell Differentiation
Transplantation

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

これを引用

Stanzel, B. V., Espana, E. M., Grueterich, M., Kawakita, T., Parel, J. M., Tseng, S. C. G., & Binder, S. (2005). Amniotic membrane maintains the phenotype of rabbit retinal pigment epithelial cells in culture. Experimental Eye Research, 80(1), 103-112. https://doi.org/10.1016/j.exer.2004.06.032

Amniotic membrane maintains the phenotype of rabbit retinal pigment epithelial cells in culture. / Stanzel, Boris V.; Espana, Edgar M.; Grueterich, Martin; Kawakita, Tetsuya; Parel, Jean Marie; Tseng, Scheffer C G; Binder, Susanne.

:: Experimental Eye Research, 巻 80, 番号 1, 01.2005, p. 103-112.

研究成果: Article

Stanzel, BV, Espana, EM, Grueterich, M, Kawakita, T, Parel, JM, Tseng, SCG & Binder, S 2005, 'Amniotic membrane maintains the phenotype of rabbit retinal pigment epithelial cells in culture', Experimental Eye Research, 巻. 80, 番号 1, pp. 103-112. https://doi.org/10.1016/j.exer.2004.06.032
Stanzel, Boris V. ; Espana, Edgar M. ; Grueterich, Martin ; Kawakita, Tetsuya ; Parel, Jean Marie ; Tseng, Scheffer C G ; Binder, Susanne. / Amniotic membrane maintains the phenotype of rabbit retinal pigment epithelial cells in culture. :: Experimental Eye Research. 2005 ; 巻 80, 番号 1. pp. 103-112.
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