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S. G. Remington, J. M. Crow, J. D. Nelson; FGF2 Enhanced Lentoid Formation in Chicken Ciliary Body Cultures. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4368.
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Lentoids (clusters of lens fiber-like cells) form in cultures of embryonic chicken ciliary body. We investigated the effects of FGF2 media supplementation on lentoid formation.
Embryonic (E6-E8) chicken ciliary body cells were dissociated with dispase II and plated on plastic dishes in M199, 5% fetal calf serum, 5% chicken serum and 1% penicillin/streptomycin/amphotericin B. Media changes were supplemented with human FGF2 (20-100 ng/ml) 1-6 times. Media was changed every 2-3 days for 5-8 weeks. Cultures were processed for immunofluorescence or Western blots with antibodies to chicken fiber cell proteins AQP0 and filensin. Δ-crystallin was monitored in Coomassie stained gels.
Lentoids formed in FGF2-supplemented ciliary body cultures after 3-5 weeks, constituting a sub-population of cells. More FGF2 treatments (5-6) and higher concentrations of FGF2 (50-100ng) resulted in the formation of more lentoids and the production of higher levels of the fiber cell proteins Δ-crystallin, AQP0 and filensin. In lentoid-rich ciliary body cultures, the levels of Δ-crystallin, AQP0 and filensin per 100 mg culture protein were approximately 50% of the levels per 100 mg embryonic chicken lens protein. Few or no lentoids formed in untreated control ciliary body cultures; AQP0 and filensin were not detected in Western blots of control ciliary body cultures.
FGF2 supplementation enhanced the formation of lentoids (containing lens fiber cell proteins) in embryonic chicken ciliary body cultures.
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