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Mitra Alizadeh, Claire M. Gelfman, Shellie R. Bench, Leonard M. Hjelmeland; Expression and Splicing of FGF Receptor mRNAs during ARPE-19 Cell Differentiation In Vitro. Invest. Ophthalmol. Vis. Sci. 2000;41(8):2357-2362.
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purpose. The expression and alternative splicing of the four FGF receptor (FGFR)
mRNAs are regulated in a developmental- and tissue-specific fashion.
Capability of differentiation in vitro of the retinal pigment
epithelial cell line ARPE-19 has been previously demonstrated. In this
study, the hypothesis that FGF receptor gene expression and the
alternative splicing of the FGFR1 mRNA is regulated as a function of
ARPE-19 differentiation in vitro was tested.
methods. ARPE-19 cells were plated at sparse or confluent densities and
maintained in culture up to 14 months. The expression of FGF receptors
and the ratio of the FGFR1β to FGFR1α splice variants of the FGFR1
transcript were quantified by a published PCR technique. Two in vivo
samples of human RPE served as controls.
results. Sparse cultures of ARPE-19 cells predominantly express FGFR1. When
these cultures are allowed to differentiate, FGFR2 is also expressed.
Samples of mRNA from RPE cells in vivo exhibit FGFR1 and FGFR2
expression as well as FGFR3 expression, a form that is minimally
apparent in vitro. The ratio of the FGFR1β to FGFR1α splice variant
decreases as a function of cell differentiation in vitro and approaches
the ratio observed in human RPE cells in vivo. Stimulation of cultures
in vitro with FGF2 as a prototypical differentiation agent does not
regulate the ratio of the FGFR1β to FGFR1α splice variant.
conclusions. Differentiation of the ARPE-19 cell line in vitro recapitulates many
but not all the in vivo patterns of FGFR expression and splicing. This
in vitro system may be useful for selected studies on how cellular
differentiation regulates FGF receptor gene expression and
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