Short-term organ culture of the retinal pigment epithelium in microtitration plates: Ultrastructural studies

  • N. M. McKechnie
  • W. A. Keegan
  • C. A. Converse
  • W. S. Foulds
Article

Abstract

Microtitration plates were used to culture simultaneously multiple, small (6 mm diameter) explants of bovine retinal pigment epithelium (RPE). Evaluation of tissue by light microscopy and by scanning and transmission electron microscopy after various incubation periods up to 6 h showed that RPE maintained in this system retains near normal morphology. Initially, the explanted RPE lacks apical microvilli, but during the first 2–3 h in culture recovery of apical microvilli occurs. The results suggest that the system is suitable for short-term maintenance of RPE for experimental purposes. Moreover, the ability to culture up to 16 explants from one bovine eye aids statistical evaluation of RPE behaviour under varying experimental conditions.

Keywords

Public Health Microscopy Electron Microscopy Transmission Electron Microscopy Light Microscopy 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Berman ER, Schwell H, Feeney L (1974) The retinal pigment epithelium. Chemical composition and structure. Invest Ophthalmol 13:675–687Google Scholar
  2. Chaitin MH, Hall MO (1983) Defective ingestion of rod outer segments by cultured dystrophic rat pigment epithelial cells. Invest Ophthalmol Vis Sci 24:812–820Google Scholar
  3. Del Monte MA, Maumenee IH (1981) In vitro culture of human retinal pigment epithelium for biochemical and metabolic study. Vision Res 21:137–142Google Scholar
  4. Droz B (1963) Dynamic condition of proteins in the visual cells of rats and mice as shown by radioactography with labeled amino acids. Anat Rec 145:157–167Google Scholar
  5. Edwards RB (1977) Cultures of rat pigment epithelium. In Vitro 13:301–304Google Scholar
  6. Edwards RB (1983) The use of tissue culture techniques to study normal and diseased retinal pigment epithelium. Prog Retinal Res 2:51–66Google Scholar
  7. Feeney L, Mixon RL (1976) An in vitro model of phagocytosis in bovine and human retinal pigment epithelium. Exp Eye Res 22:533–548Google Scholar
  8. Flood MT, Gouras P (1981) The organization of human retinal pigment epithelium in vitro. Vision Res 21:119–126Google Scholar
  9. Flood MT, Gouras P, Kjeldbye H (1980). Growth characteristics and ultrastructure of human retinal pigment epithelium in vitro. Invest Ophthalmol Vis Sci 19:1309–1320Google Scholar
  10. Goldman AI, O'Brien PJ (1978) Phagocytosis in the retinal pigment epithelium of the RCS rat. Science 201:1023–1025Google Scholar
  11. Hall MO (1978) Phagocytosis of light and dark adapted rod outer segments by cultured pigment epithelium. Science 202:526–528Google Scholar
  12. Hayashi M, Matsumoto A, Hamashima Y, Tsukahara I (1979) Phagocytic activity of cultured retinal pigment epithelium. Uptake of polystyrene shperes andStaphlococcus aureus. Exp Eye Res 28:427–434Google Scholar
  13. McKechnie NM, Keegan WA, Converse CA, Foulds WS (1985) Glycosaminoglycans: their effect on rod outer segment phagocytosis by bovine retinal pigment epithelium explants. Invest Ophthalmol Vis Sci [Suppl] 26:196Google Scholar
  14. Mason WT, Fager RS, Abrahamson EW (1973) Ultrastructure of the receptor and epithelial layers of the bovine retina. J Anat 115:289–308Google Scholar
  15. Masterson E, Goldman AI, Chader GJ (1981) Phagocytosis of rod outer segments by cultured epithelial cells. Vision Res 21:143–145Google Scholar
  16. Mullen RJ, LaVail MM (1976) Inherited retinal dystrophy: a primary defect in pigment epithelium determined with experimental rat chimeras. Science 192:799–801Google Scholar
  17. Orenstein JM, Shelton E (1977) Membrane phenomena accompanying erythrophagocytosis. A scanning electron microscopy study. Lab Invest 36:363–374Google Scholar
  18. Philp NJ, Bernstein MH (1981) Phagocytosis by retinal pigment epithelium explants in culture. Exp Eye Res 33:47–53Google Scholar
  19. Silverstein SC, Steinman RM, Sohn ZA (1977) Endocytosis. Ann Rev Biochem 46:669–722Google Scholar
  20. Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43Google Scholar
  21. Steinman RM, Brodie SE, Cohn ZA (1976) Membrane flow during pinocytosis. A stereological analysis. J Cell Biol 68:665–687Google Scholar
  22. Tamai M, Mizuno K, Chader GJ (1982) In vitro studies on shedding and phagocytosis or rod outer segments in the rat retina: effect of oxygen concentration. Invest Ophthalmol Vis Sci 22:439–448Google Scholar
  23. Tso MOM, Albert D, Zimmerman LE (1973) Organ culture of human RPE and choroid: a model for the study of cytologic behavior of RPE in vitro. Invest Ophthalmol 12:554–566Google Scholar
  24. Young RW (1965) Renewal of photoreceptor outer segments. Anat Rec 151:484Google Scholar
  25. Young RW (1967) The renewal of photoreceptor cell outer segments. J Cell Biol 33:61–72Google Scholar
  26. Young RW (1977) Visual cell renewal systems and the problem of retinitis pigmentosa. In: Landers MB, III, Wolbarsht ML, Dowling JE, Laties AM (eds) Retinitis pigmentosa. Clinical implications of current research. Plenum Press, New York, pp 93–113Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • N. M. McKechnie
    • 1
  • W. A. Keegan
    • 2
  • C. A. Converse
    • 2
  • W. S. Foulds
    • 1
  1. 1.Department of OphthalmologyUniversity of GlasgowScotland
  2. 2.Department of PharmacyUniversity of StrathclydeGlasgowScotland

Personalised recommendations