Descemet's membrane substrate from human donor lens anterior capsule.

BACKGROUND

To study the potential use of human donor anterior lens capsule as a Descemet's membrane substrate.

METHODS

Anterior lens capsules were recovered from the lenses of 30 cornea donors. Human corneal endothelial cells were recovered from the remaining corneal sclera rims of 15 donor corneas used for penetrating keratoplasty. Samples were sorted into three groups. Group 1 consisted of 10 samples in which the endothelial cells were allowed to grow on anterior lens capsules. In Group 2 human corneal endothelial cells grew on a collagen membrane and in Group 3 on polystyrene culture plates. Cell density, morphology and adherence of the cell-capsule complex were evaluated at 1, 4, 7 and 14days with a phase-contrast microscope, a scanning electron microscope and by histology. Cell viability was quantified by a microscopic live-dead assay. Expression of zonula occludens-1, Na(+) /K(+) -adenosine triphosphatase, tissue transglutaminase and vimentin were investigated by immunohistochemistry.

RESULTS

A mean diameter of 10.05±0.13mm of anterior capsule was obtained as a substrate for cell culture. Endothelial cell density of Group 1 was measured at 2455.4±283.8cells/mm(2) , which was also comparable with the cell density of the control group. Cell viability was 95% or superior in all groups and multiple cellular interconnections developed between growing cells. Immunohistochemical analysis demonstrated strongly positive staining for all investigated proteins. Electron microscopy confirmed the adherence and monolayer growth of the endothelial cells.

CONCLUSIONS

Human donor anterior lens capsule might therefore be a potential scaffold for the ex vivo expansion of human corneal endothelial cells.