Abstract
Purpose :
Expansion of the conjunctiva ex vivo, for study and transplantation, containing both epithelial and goblet cells is difficult (Marsh et al. Exp Eye Res. 2002;74:61-69). Biomaterials surfaces are known to control cellular processes such as adhesion and differentiation and this has been demonstrated with Biomer Technology Ltd.’s novel line of acrylic polymers (Glennon-Alty et al. Acta Biomat. 2013;9:6041-6051). These acrylic surfaces have been designed with specific surface chemical functionality to mimic extracellular matrix molecules. We have evaluated epithelial and goblet cell growth on the Biomer acrylics to determine optimal culture surfaces for both cell types.
Methods :
Metabolic activity and cell number of conjunctival (HCjE-Gi) and goblet (HT29 MTX) cell lines were investigated, using AlamarBlue and CyQuant assays respectively, on 8 Biomer acrylics over time (N=3). Cells were stained for CK19 and phalloidin to view morphology.
Results :
Epithelial cells grown on ESP003, ESP008, ESP010, ESP012 and BLT015 had a greater metabolic activity relative to control at 24hrs. By day 7 cells on all materials had lower metabolic activity relative to controls. ESP011 and ESP012 had the highest relative metabolic activity at day7 of 0.85±0.01 and 0.82±0.01 respectively. The number of epithelial cells, from DNA measurements, on ESP011 and ESP012 at day 7 was 23,305±9,117 and 24,164±5,315 respectively; and CyQuant assays demonstrated that the numbers of cells on ESP007 and BTL015 were also greater than on controls. Only ESP003, ESP012 and BTL015 supported goblet cell growth to day 7. Metabolic activities of cells on these materials on day 7, relative to controls, were 0.53±0.44, 0.57±0.43 and 0.88±0.65 respectively. The number of cells on substrates that supported goblet cell growth at day 7; ESP003, ESP012 and BTL015, were 331,490±144,433, 276,642±152,168 and 374,641±198,172 respectively. HCjE cells on ESP0003, ESP007, ESP011, ESP012, and BTL015 had an epithelial morphology.
Conclusions :
ESP003, ESP012 and BTL015 supported the growth of both cell types. Adhesion and growth was greater for goblet cells compared to epithelial cells on these materials. The improved adhesion of goblet cells on these materials may allow a combined co-culture with good conjunctival properties, to be evaluated with primary conjunctival cultures.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.