Abstract
Purpose: :
To investigate growth characteristics of lens epithelial cells from human capsular explants in response to substrates and medium variants, which included electrospun nanofibres and various growth stimulating factors.
Methods: :
Human capsulorhexis specimen from cataract surgery where cultured in 1) Media 199 supplemented with 10% fetal calf serum (FCS) and hydrocortisone (HC), or 2) the retinal pigment epithelium (RPE) specific serum reduced medium by Hu& Bok containing bovine retina extract (BRE). Cells were grown on lens capsules, gelatine coated glas slides, plastic or electrospun polyamide nanofibres. Their growth behaviour was monitored by phase contrast microscopy and with immunofluorescence for vimentin, smooth muscle actin (SMA), f-actin, filensin, -tubulin, endoglin and basic fibroblast growth factor (FGF-2).
Results: :
Cell growth was independent of donor age of the capsulorhexis specimen. Both gelatine coating as well as electrospun nanofibres stimulated epithelial cell growth with diverging cell morphologies. RPE specific medium proved to be a pronounced stimulator of epithelial growth independent of the substrate, but as opposed to FCS it stimulated multilayered growth. F-actin, SMA, vimentin and -tubulin staining demonstrated more stress fibre patterns in epithelial cells growing outside the original capsular substrate. In contrast, Filensin staining was localized at cell membranes on all substrates. FGF-2 expression was always membrane bound and showed a vanishing signal gradient with increasing distance from the capsule.
Conclusions: :
Cultured human lens epithelial cells are responsive to substrates variants and specific growth stimulators. This preserved in vitro reactivity could make them suitable for various aspects of tissue engineering in the eye.
Keywords: cytoskeleton • immunohistochemistry • growth factors/growth factor receptors