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G. Astrid Limb, Thomas E. Salt, Peter M. G. Munro, Stephen E. Moss, Peng T. Khaw; In Vitro Characterization of a Spontaneously Immortalized Human Müller Cell Line (MIO-M1). Invest. Ophthalmol. Vis. Sci. 2002;43(3):864-869.
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purpose. To characterize a spontaneously immortalized human Müller cell
line and to determine whether it retains the characteristics of primary
isolated cells without undergoing differentiation in vitro.
methods. An immortalized cell line obtained from human retina was investigated
for the expression of known markers of Müller cells, including
cellular retinaldehyde binding protein (CRALBP), glutamine synthetase,
epidermal growth factor receptor (EGF-R), α-smooth muscle actin
(α-SMA), and glial fibrillary acidic protein (GFAP). Also examined
were the morphologic features of these cells, by scanning and
transmission electron microscopy, and their functional characteristics,
by electrogenic responses to glutamate. In addition, comparative
studies were made of these cells with primary cultures of freshly
isolated human Müller cells.
results. The cells expressed CRALBP, EGF-R, glutamine synthetase, and α-SMA,
as judged by confocal microscopy and Western blot analysis of cell
lysates. Western blot analysis did not detect GFAP in cell lysates, but
confocal microscopy showed that occasional cells expressed GFAP after
detachment from the monolayer. The morphologic features of the cells
examined, as judged by scanning and transmission electron microscopy,
resemble those of cells derived from primary cell cultures. They
possess villous projections on their apical surfaces and contain loose
bundles of microtubules aligned parallel to one another and the long
axis of the cell process. Characteristically, they contain abundant
deposits of glycogen particles that do not differ from those seen in
primary isolated cells. Preliminary recordings with intracellular
electrodes revealed that these cells have properties similar to those
described for mammalian Müller cells and depolarize in response
to l-glutamate without significant change in membrane
resistance, consistent with the well-established electrogenic uptake of
this amino acid.
conclusions. A spontaneously immortalized Müller cell line was characterized
that retains the characteristics of primary isolated cells in culture.
To the authors’ knowledge, it constitutes the first human Müller
cell line reported in the literature. It has been named MIO-M1
(Moorfields/Institute of Ophthalmology-Müller 1) after the
authors’ institution. Availability of this human cell line will
facilitate studies designed to obtain a better understanding of the
role of Müller cells in normal and pathologic
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