Ex vivo cultivation of corneal limbal epithelial cells in a thermoreversible polymer (Mebiol Gel) and their transplantation in rabbits: an animal model.
We evaluated the efficacy of autologous expanded corneal epithelial cell transplants derived from harvested limbal biopsy cultured on a thermoreversible polymer (Mebiol Gel) for the management of unilateral limbal stem cell deficiency (LSCD). Corneal limbal biopsies from 12 rabbits were cultured on a thermoreversible polymer Mebiol Gel at 37 degrees C. Cells were harvested from the dishes after 3 weeks by reducing temperature to 4 degrees C. Autologous transplantation was undertaken to reconstruct the experimentally induced limbal stem cell deficiency in the rabbit eyes. The corneas of both eyes of all rabbits were harvested later for molecular studies. Reparative surgery was a total success in seven rabbits, partial success in two, and failure in three eyes. Histology of the seven successful eyes showed the successful growth of the corneal epithelium. Immunohistochemistry and reverse transcriptase polymerase chain reaction showed the cornea phenotype and stem cell-associated markers in the limbus of the seven successful eyes, indicating the homing of these cells into limbus. In the three failure cases and in the two control rabbit eyes, used in the study, histology showed presence of goblet cells and vascularization in the stroma with abortive formation of corneal epithelium. Our results suggest that transplantation of autologous limbal epithelial cells grown in thermoreversible polymer Mebiol Gel may restore a nearly normal ocular epithelial surface in eyes with unilateral LSCD.
Sitalakshmi
G
G
Department of Cornea, Vision Research Foundation, Chennai, India.
Sudha
B
B
Madhavan
H N
HN
Vinay
S
S
Krishnakumar
S
S
Mori
Yuichi
Y
Yoshioka
Hiroshi
H
Abraham
Samuel
S
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D003316:Corneal Diseases / Q000473:pathology; D004847:Epithelial Cells / Q000187:drug effects* / Q000473:pathology / Q000637:transplantation*; D019573:Epithelium, Corneal / Q000473:pathology*; D015353:Eye Enucleation; D005782:Gels; D016850:Limbus Corneae / Q000473:pathology*; D008297:Male; D023421:Models, Animal; D011108:Polymers / Q000494:pharmacology*; D012313:RNA / Q000302:isolation & purification; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013499:Surface Properties / Q000187:drug effects; D013696:Temperature
D015415:Biomarkers; D005782:Gels; D011108:Polymers; D012313:RNA