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"Immortalized cells" are cells with extended lifespans that retain characteristics of low-passage or freshly dissociated cells (contact inhibition, substrate-dependent growth, but no senescence or culture-related phenotypic changes after 8 to 10 passages)
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"Immortalized cells" are cells with extended lifespans that retain characteristics of low-passage or freshly dissociated cells (contact inhibition, substrate-dependent growth, but no senescence or culture-related phenotypic changes after 8 to 10 passages).
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9
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Viral producer cell line PA317LXSN 16E6E7 (American Type Culture Collection) was used
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C. L. Halbert, G. W. Demers, D. A. Galloway, J. Virol. 65, 473 (1991); J. Virol. 66, 2125 (1992). Viral producer cell line PA317LXSN 16E6E7 (American Type Culture Collection) was used.
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0345669035
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Cytogenetic analyses were performed by H. S. Wang (Cytogenetics Laboratory, Children's Hospital of Eastern Ontario, Ottawa) using routine G-banding techniques with 400-to 450-band resolution
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Cytogenetic analyses were performed by H. S. Wang (Cytogenetics Laboratory, Children's Hospital of Eastern Ontario, Ottawa) using routine G-banding techniques with 400-to 450-band resolution.
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19
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0344374355
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TeloQuant Telomeric Repeat Amplification Protocol kit (Pharmingen Canada)
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TeloQuant Telomeric Repeat Amplification Protocol kit (Pharmingen Canada).
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Epithelial cells showed corneal epithelial-specific keratin 12 staining [A. Schermer, S. Galvin, T. T. Sun, J. Cell Biol. 103, 49 (1986)] with AE5 antibody (ICN Biochemicals, Aurora, OH). Stromal cells stained for vimentin (Roche Diagnostics, Montreal). Endothelial cells expressed α2(VIII) collagen mRNA [Y. Muragaki et al., J. Biol. Chem. 266, 7721 (1991)] and lacked keratin 12 staining.
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Epithelial cells showed corneal epithelial-specific keratin 12 staining [A. Schermer, S. Galvin, T. T. Sun, J. Cell Biol. 103, 49 (1986)] with AE5 antibody (ICN Biochemicals, Aurora, OH). Stromal cells stained for vimentin (Roche Diagnostics, Montreal). Endothelial cells expressed α2(VIII) collagen mRNA [Y. Muragaki et al., J. Biol. Chem. 266, 7721 (1991)] and lacked keratin 12 staining.
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Cell lines producing successful constructs were from young donors, were immortalized with HPV E6/E7, and had appropriate electrophysiological responses
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Cell lines producing successful constructs were from young donors, were immortalized with HPV E6/E7, and had appropriate electrophysiological responses.
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27
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0345237167
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in preparation
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M. Griffith et al., in preparation.
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unpublished data
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D. H. Geroski, J. C. Kies, and H. F. Edelhauser [Curr. Eye Res. 3, 331 (1984)] showed that 100 μM ouabain causes swelling of human corneas (41 μm per hour); after 2 hours corneas would swell by 82 μm (16%).
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34
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0345237166
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note
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Constructs, with epithelium sealed with silicone oil to prevent movement of water across the anterior surface, were measured using OCT (Zeiss-Humphrey, San Leandro, CA). The average change in thickness was calculated from four to six measurements before and after treatment of the endothelial surface for 2 hours with 100 μM ouabain or Dulbecco's modified Eagle's medium alone (DMEM, Gibco).
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36
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0000021897
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c-fos: Y. Okada et al., Graefes Arch. Clin. Exp. Ophthalmol. 236, 853 (1998); IL-1 and IL-6: C. Sotozono et al., Curr. Eye Res. 16, 670 (1997); bFGF; V. P. T. Hoppenreijs, E. Pels, G. F. J. M. Vrensen, W. F. Treffers, Invest. Ophthalmol. Visual Sci. 35, 931 (1994); VEGF: S. Amano, R. Roban, M. Kuroki, M. Tolentino, A. P. Adamis, Invest. Ophthalmol. Visual Sci. 39, 18 (1998); Coll I: W. J. Power, A. H. Kaufman, J. Merayo-Lloves, V. Arrunategui-Correa, C. S. Foster, Curr. Eye Res. 14, 879 (1995).
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c-fos: Y. Okada et al., Graefes Arch. Clin. Exp. Ophthalmol. 236, 853 (1998); IL-1 and IL-6: C. Sotozono et al., Curr. Eye Res. 16, 670 (1997); bFGF; V. P. T. Hoppenreijs, E. Pels, G. F. J. M. Vrensen, W. F. Treffers, Invest. Ophthalmol. Visual Sci. 35, 931 (1994); VEGF: S. Amano, R. Roban, M. Kuroki, M. Tolentino, A. P. Adamis, Invest. Ophthalmol. Visual Sci. 39, 18 (1998); Coll I: W. J. Power, A. H. Kaufman, J. Merayo-Lloves, V. Arrunategui-Correa, C. S. Foster, Curr. Eye Res. 14, 879 (1995).
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Sotozono, C.1
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c-fos: Y. Okada et al., Graefes Arch. Clin. Exp. Ophthalmol. 236, 853 (1998); IL-1 and IL-6: C. Sotozono et al., Curr. Eye Res. 16, 670 (1997); bFGF; V. P. T. Hoppenreijs, E. Pels, G. F. J. M. Vrensen, W. F. Treffers, Invest. Ophthalmol. Visual Sci. 35, 931 (1994); VEGF: S. Amano, R. Roban, M. Kuroki, M. Tolentino, A. P. Adamis, Invest. Ophthalmol. Visual Sci. 39, 18 (1998); Coll I: W. J. Power, A. H. Kaufman, J. Merayo-Lloves, V. Arrunategui-Correa, C. S. Foster, Curr. Eye Res. 14, 879 (1995).
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Hoppenreijs, V.P.T.1
Pels, E.2
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Treffers, W.F.4
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41
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0031965601
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c-fos: Y. Okada et al., Graefes Arch. Clin. Exp. Ophthalmol. 236, 853 (1998); IL-1 and IL-6: C. Sotozono et al., Curr. Eye Res. 16, 670 (1997); bFGF; V. P. T. Hoppenreijs, E. Pels, G. F. J. M. Vrensen, W. F. Treffers, Invest. Ophthalmol. Visual Sci. 35, 931 (1994); VEGF: S. Amano, R. Roban, M. Kuroki, M. Tolentino, A. P. Adamis, Invest. Ophthalmol. Visual Sci. 39, 18 (1998); Coll I: W. J. Power, A. H. Kaufman, J. Merayo-Lloves, V. Arrunategui-Correa, C. S. Foster, Curr. Eye Res. 14, 879 (1995).
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Amano, S.1
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c-fos: Y. Okada et al., Graefes Arch. Clin. Exp. Ophthalmol. 236, 853 (1998); IL-1 and IL-6: C. Sotozono et al., Curr. Eye Res. 16, 670 (1997); bFGF; V. P. T. Hoppenreijs, E. Pels, G. F. J. M. Vrensen, W. F. Treffers, Invest. Ophthalmol. Visual Sci. 35, 931 (1994); VEGF: S. Amano, R. Roban, M. Kuroki, M. Tolentino, A. P. Adamis, Invest. Ophthalmol. Visual Sci. 39, 18 (1998); Coll I: W. J. Power, A. H. Kaufman, J. Merayo-Lloves, V. Arrunategui-Correa, C. S. Foster, Curr. Eye Res. 14, 879 (1995).
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Power, W.J.1
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43
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0345237164
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Live/Dead Viability/Cytotoxicity kit (Molecular Probes, Eugene, OR)
-
Live/Dead Viability/Cytotoxicity kit (Molecular Probes, Eugene, OR).
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46
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0345669031
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note
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Supported by grants from the Natural Sciences and Engineering Research Council of Canada, Medical Research Council-Pharmaceutical Manufacturers' Association of Canada, and the P&G International Program for Animal Alternatives. We thank S. Whelan and P. Berg for AdS and psvBneo plasmids, C. Smith and Ottawa Hospital Research Institute colleagues, and E. Hay and A. Tashjian Jr. for helpful insights during preparation of this manuscript.
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