Abstract
Purpose: :
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.
Methods: :
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.
Results: :
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.
Conclusions: :
FGF2 supplementation enhanced the formation of lentoids (containing lens fiber cell proteins) in embryonic chicken ciliary body cultures.
Keywords: ciliary body • differentiation • growth factors/growth factor receptors