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Eleanor A. Blakely, Kathleen A. Bjornstad, Polly Y. Chang, Morgan P. McNamara, Edmund Chang, Geraldine Aragon, Serena P. Lin, GeMing Lui, Jon R. Polansky; Growth and Differentiation of Human Lens Epithelial Cells In Vitro on Matrix. Invest. Ophthalmol. Vis. Sci. 2000;41(12):3898-3907.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To characterize the growth and maturation of nonimmortalized
human lens epithelial (HLE) cells grown in vitro.
methods. HLE cells, established from 18-week prenatal lenses, were maintained on
bovine corneal endothelial (BCE) extracellular matrix (ECM) in medium
supplemented with basic fibroblast growth factor (FGF-2). The identity,
growth, and differentiation of the cultures were characterized by
karyotyping, cell morphology, and growth kinetics studies, reverse
transcription–polymerase chain reaction (RT-PCR), immunofluorescence,
and Western blot analysis.
results. HLE cells had a male, human diploid (2N = 46) karyotype. The
population-doubling time of exponentially growing cells was 24 hours.
After 15 days in culture, cell morphology changed, and lentoid
formation was evident. Reverse transcription–polymerase chain reaction
(RT-PCR) indicated expression of αA- and βB2-crystallin, fibroblast
growth factor receptor 1 (FGFR1), and major intrinsic protein (MIP26)
in exponential growth. Western analyses of protein extracts show
positive expression of three immunologically distinct classes of
crystallin proteins (αA-, αB-, and βB2-crystallin) with time in
culture. By Western blot analysis, expression of p57KIP2, a
known marker of terminally differentiated fiber cells, was detectable
in exponential cultures, and levels increased after confluence. MIP26
and γ−crystallin protein expression was detected in confluent
cultures, by using immunofluorescence, but not in exponentially growing
conclusions. HLE cells can be maintained for up to 4 months on ECM derived from BCE
cells in medium containing FGF-2. With time in culture, the cells
demonstrate morphologic characteristics of, and express protein markers
for, lens fiber cell differentiation. This in vitro model will be
useful for investigations of radiation-induced cataractogenesis and
other studies of lens toxicity.
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