Purchase this article with an account.
J. Moreno-Montanes, A. Fernandez-Hortelano, M. Garcia-Guzman, D. Lozano, J. Echeveste, F. Prosper; Transplantation of Autologous Limbal Stem Cells Culture on Human Amniotic Membrane as Treatment of Complete Limbal Stem Cell Deficiency in a Rabbit Model. Invest. Ophthalmol. Vis. Sci. 2007;48(13):468.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To study the outcome of autologous limbal stem cell (LSC) culture on human denuded amniotic membrane as a treatment for corneal regeneration in a total limbal stem cell deficiency animal model.
Complete limbal stem cell deficiency (LSCD) was induced by treatment with n-heptanol in thirty rabbits. After two months animals underwent transplantation of autologous limbal stem cell culture on human denuded amniotic membrane (n=15, Group I) or received symptomatic treatment (n=15, Group II). Culture explants were culture by a combined method including a first part on plastic for 14-21 days and secondly on denuded amniotic membrane (72 hours). Transplanted cells were characterized by immunohistochemistry using antibodies against p63, CK3/12 and CK14. Clinical outcome was based on biomicroscopy examination and impression cytology and was performed monthly in both groups. After 6 months animals were sacrificed and the eyes subjected to histological and IHQ examination.
6 months post-treatment, histological examination showed complete corneal epithelium regeneration and a significant decrease of goblet cells in the group of animals treated with LSC on AM. Immunohistochemistry revealed a normal distribution of corneal epithelial markers CK3/12 and CK14. Clinical outcome also showed significant corneal epithelization and reduced inflammation and neovascularization on LSCs transplantation group. Corneal ulceration was also resolved on LSCs transplantation group.
Our results suggest that ex vivo expansion of LSCs can be achieved by using a combined culture method on plastic and denuded human amniotic membrane. Such expanded LSCs can successfully reconstruct corneal surface and improve clinical and histological outcome of total LSCD. Comparing to a control group, treatment with transplantation of LSCs cultured on amniotic membrane improves clinical inflammation and corneal surface regeneration and decreases number of Goblet cells.
This PDF is available to Subscribers Only