Abstract
The retinal pigment epithelium (RPE) is a layer of multipotential cells of neural ectoderm origin lying between Bruch’s membrane and the neural retina. The RPE subserves several essential ocular functions, including phagocytosis of shed photoreceptor outer segments, maintenance of the blood-retinal barrier, absorption of stray light, regulation of the biochemical, metabolic, and ionic composition of the subretinal space, and induction of embryonic differentiation of adjacent neural retina and choroids (1). Experimental evidence indicates that early in embryonic life the neural retina can regenerate from the pigment epithelium (2). In vitro cultures of pure RPE provide a vehicle for studying RPE function in both normal and diseased states, and may also serve as a model for other neural cells (3,4). Multiple techniques have been described for culturing human RPE ((5) – (12)). The authors describe here a modification of the technique of DelMonte and Maumenee (10), which is simple and effective in establishing primary cultures and extended cell lines of human RPE for research.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Zinn, K. M. and Benjamin-Henkind, J. (1991) Retinal pigment epithelium, in Biomedical Foundations of Ophthalmology, vol. I (Duane, T. D., ed.), Harper and Row, Philadelphia, PA, ch. 21, pp. 1–20.
Coulombre, J. L. and Coulombre, H. A. (1965) Regeneration of neural retina from the pigmented epithelium in the chick embryo. Dev. Biol. 17, 79–83.
Newsome, D. A. (1983) Retinal pigmented epithelium culture: current applications. Trans. Ophthalmol. Soc. UK 103, 458–466.
Boulton, M. E., Marshall. J., and Mellerio, J. (1982) Human retinal pigment epithelial cells in tissue culture: a means of studying inherited retinal diseases. Birth Defects 19, 101–118.
Albert, D. M. and Buyukmihci, N. (1979) Tissue culture of the retinal pigment epithelium, in The Retinal Pigment Epithelium (Zinn, K. M. and Marmour, M. F., eds.), Harvard Univ. Press, Cambridge, MA, ch. 16, pp. 277–292.
Albert, D. M., Tso, M. O., and Rabson, A. S. (1972) In vitro growth of pure cultures of retinal pigment epithelium. Arch. Ophthalmol. 88, 63–69.
Mannaugh, J., Dhaarmendra, V. A., and Irvine, A. R. (1973) Tissue culture of human retinal pigment epithelium. Invest. Opthalmol. Vis. Sci. 12, 52–64.
Edwards, R. B. (1982) Culture of mammalian retinal pigment epithelium and neural retina. Methods Enzymol. 81, 39–43.
Flood, M. T., Gouras, P., and Kjeldbye, H. (1980) Growth characteristics and ultra-structure of human retinal pigment epithelium in vitro. Invest. Opthalmol. Vis. Sci. 19, 1309–1320.
DelMonte, M. A. and Maumenee, I. H. (1981) In vitro culture of human retinal pigment epithelium for biochemical and metabolic study. Vis. Res. 21, 137–142.
Pfeffer, B. A., Clark, V. M. Flanery, J. G., and Bok, D. (1986) Membrane receptors for retinal-binding protein in cultured human retinal pigment epithelium. Invest. Ophthalmol. Vis. Sci. 27, 1031–1040.
Mircheff, A. K., Miller, S. S., Farber, D. B., Bradley M. E., O’Day. W. T., and Bok, D. (1990) Isolation and provisional identification of plasma membrane populations from cultured human retinal pigment epithelium. Invest. Opthalmol. Vis. Sci. 31, 863–878.
DelMonte, M. A. and Maumenee, I. H. (1980) New techniques for in vitro culture of human retinal pigment epithelium, in Birth Defects: Original Article Series, vol. XVI, no. 2, pp. 327–338.
Dutt, K., Scott, M. M., Del Monte, M., et al. (1991) Extracellular matrix mediated growth and differentiation in human pigment epithelial cell line 0041. Curr. Eve Res. 10, 1089–1100.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Stevens, M.J., Larkin, D.D., Feldman, E.L., DelMonte, M.A., Greene, D.A. (2005). Establishment, Maintenance, and Transfection of In Vitro Cultures of Human Retinal Pigment Epithelium. In: Picot, J. (eds) Human Cell Culture Protocols. Methods in Molecular Medicine™, vol 107. Humana Press. https://doi.org/10.1385/1-59259-861-7:343
Download citation
DOI: https://doi.org/10.1385/1-59259-861-7:343
Publisher Name: Humana Press
Print ISBN: 978-1-58829-222-3
Online ISBN: 978-1-59259-861-8
eBook Packages: Springer Protocols