Abstract
• Background: We report a modified method for the isolation and propagation of adult human Müller cells in culture. • Methods: The retina of postmortem human eyes was mechanically dissociated and cultured. Using immunocytochemical techniques, these cells were stained with monoclonal antibodies specific for Müller cells, glial fibrillary acidic protein (GFAP), vimentin, glutamine synthetase (GS) and keratin. Transmission electron microscopy (TEM) was also performed. • Results: The dissociated and cultured cells expressed vimentin and GS, but not GFAP. At least 85% of these cells stained with a Müller tell-specific monoclonal antibody. Using TEM, flat cells containing 13-nm intermediate filaments and glycogen were identified. • Conclusion: Human retinal Müller cells tan be isolated and propagated in culture. Purified cell cultures are required for controlled studies of the normal physiology and pathologie responses of Müller cells.
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Adamis AP, Miller JW, Bernal M, D' Amico DJ, Folkman J, Yeo T, Yeo K (1994) Increased vascular endothelial growth factor levels in the vitreous of eyes with proliferative diabetic retinopathy. Am J Ophthaolmol 118:445–450
Barbour B, Brew H, Attwell D (1988) Electrogenic glutamate uptake in glial cells is activated by intracellular potassium. Nature 335:433–435
Chan C-C, Rozenszajn LA, Nussenblatt RB, Muellenberg-Coulombre C, Hsu S, Palestine AG, Lando Z, Ben-Ezra D (1984) Monoclonal antibodies to Müller cells of the retina. Invest Opthalmol Vis Sci 25:1007–1012
Cunha-Vaz JG, Shakib M, Ashton N (1966) Studies on the permeability of the blood-retinal barrier. I. On the existence, development, and site of a blood-retinal barrier. Br J Ophthalmol 50:441–453
D'Amore PA (1994) Mechanisms of retinal and choroidal neovascularization. Invest Ophthalmol Vis Sci 35:3974–3979
Hicks D, Courtois Y (1990) The growth and behavior of rat retinal Müller cells in vitro. 1. An improved method for isolation and culture. Exp Eye Res 51:119–129
Hiscott PS, Grierson I, Tromibetta CJ, Rahi AHS, Marshall J, McLeod D (1984) Retinal and epiretinal glia — an immunohistochemical study. Br J Ophthalmol 68:698–707
Hsu SM, Raine L, Fander H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comiparison between ABC and unlabeled antibody (PAP) procedures. J Histochemi Cytochem 29:577–580
Kuwabara T, Cogan DG (1961) Retinal glycogen. Arch Ophthalmol 66:680–688
Lewis PL, Kaska DD, Vaughan DK, Fisher SK (1988) An immunocytochemical study of cat retinal Müller cells in culture. Exp Eye Res 47:855–868
Meister A (1974) Glutamine synthetase of mamimals. In: Boyer PD (ed) The enzymes. Academic Press, London, pp 699–754
Newman EA (1986) The Müller cell. In: Fedoroff S, Vernadakis A (eds) Astrocytes, vol 1, Academic Press, Orlando, Fla, pp 149–171
Newman EA (1987) Distribution of potassium conductance in mamimalian Müller (glial) cells: a comparative study. J Neurosci 7:2423–2432
Puro DG, Mano T, Chan C, Fukuda M, Shimada H (1990) Thrombin stimulates the proliferativn of human retinal glial cells. Graefe's Arch Clin Exp Ophthalmol 228:169–173
Roberge FG, Caspi RR, Chan C, Kuwabara T, Nussenblatt RB (1985) Long term culture of Müller cells from adult rats in the presence of activated lymphocytes/monocytes products. Curr Eye Res 4:975–982
Sarthy PV (1985) Establishment of Müller cell cultures from adult rat retina. Brain Res 337:138–141
Savage FJ, Day JE, Hogg P, Grierson I (1988) Tissue culture of retinal glial cells. Eye 2 [Suppl]:S164–179
Tout S, Chan-Ling T, Hollander H, Stone J (1993) The role of Müller cells in the formation of the blood-retinal barrier. Neuroscience 55:291–301
Uchihori Y, Puro DG (1993) Glutamate as a neuron-to-glial signal for mitogenesis: role of glialN-methyl-d-aspartate receptors. Brain Res 613:212–220
Vinores SA, Camipochiaro PA, Conway BP (1990) Ultrastructural and electron-immunocytochemical characterization of cells in epiretinal membranes. Invest Ophthalmol Vis Sci 31:14–28
Wakakura M, Foulds WS (1988) Immunocytochemical characteristics of Müller cells cultured from adult rabbit retina. Invest Ophthalmol Vis Sci 29:892–900
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Arroyo, J.G., Ghazvini, S. & Char, D.H. An immunocytochemical study of isolated human retinal Müller tells in culture. Graefe's Arch Clin Exp Ophthalmol 235, 411–414 (1997). https://doi.org/10.1007/BF00947059
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DOI: https://doi.org/10.1007/BF00947059