Ex VivoCultivation of Corneal Limbal Epithelial Cells in a Thermoreversible Polymer (Mebiol Gel) and Their Transplantation in Rabbits: An Animal Model

Abstract

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.

Successful transportation of human corneal endothelial tissues without cool preservation in varying Indian tropical climatic conditions and in vitro cell expansion using a novel polymer

“…[13] In addition, the use of TGP as a scaffold for delivery of cells has shown better biocompatibility and engraftment without any toxicity. [11] From our study, it is evident that the micro-architecture of TGP maximizes the corneal endothelial cell viability upto 72 h of transport without using any cool preservation methodologies. Moreover, TGP helps in maintaining the cell morphology and viability with the formation of healthy cell spheres even upto 30 days in vitro .…”

“…It is transparent and the sol-gel transition temperature can be controlled by altering the chemical composition. [10] TGP has been shown to support the growth of different cell types, including corneal limbal stem cells,[11] Hepatocytes,[12] Chondrocytes,[13] continuous cell lines such as the Vero, HEp-2, HeLa, BHK-21, McCoy and CHO cell lines,[14] embryonic stem cells, and induced pluripotent stem cells. [15] Furthermore, TGP is a purely synthesized biocompatible polymer that has no biological contaminants.…”

In vitro culture and characterization of enteric neural precursor cells from human gut biopsy specimens using polymer scaffold

“…Because transplantation of neurospheres from fetal and post natal intestine derived neural crest cells has already proven to produce functional neurons in the post natal colon of mice (18), further studies asking whether TGP can be used as a carrier for NLB transplantation in aganglionic gut models to help retain the neural progenitors in the region where they should re-colonize and form neural; glial cells are warranted. Suggesting TGP as a carrier for transplantation of NLBs is based upon earlier studies in which TGP was employed for transplantation of stem cells, progenitor cells and adult cells in animal models (19)(20)(21). The present study is only a preliminary study and extensive studies are warranted before this approach could be standardized and considered for treatment of Hirschsprung's disease.…”

Towards the use of hydrogels in the treatment of limbal stem cell deficiency

“…Mebiol ® supported limbal explant proliferation and LEC cultured on this gel expressed LSC markers (ATP-binding cassette sub-family G member 2 and p63), transient amplifying cell markers (connexin 43, integrin α9) and cornea differentiation marker (CK3) [76]. The transplantation of autologous LEC grown in Mebiol ® was shown to restore a nearly normal ocular epithelial surface in eyes with unilateral LSCD in rabbit models [77], indicating that this hydrogel is almost at the pre-clinical stage.…”