Cultivation of limbal stem cells on a biopolymer carrier (preliminary study)

Abstract

BACKGROUND: Limbal stem cell deficiency is a complicated pathology of ocular surface, which is not always helped by conservative methods of treatment and surgery is limited by available sources of tissue. Consequently, searching for new effective methods of its treatment is now gaining popularity. The most promising approach is transplantation of tissue-engineered constructs consisting of cultured limbal stem cells (LSCs) and variety of biopolymer carriers. AIM: This study was performed to obtain and characterize a tissue-engineered constructs consisting of cultured LSCs and collagen membrane. MATERIALS AND METODS: The study was performed at the Krasnov Research Institute at the Krasnov Research Institute of Eye Diseases and Koltzov Institute of Developmental Biology in cooperation with IMTEK Ltd. with a series of experiments. Two Chinchilla rabbits with an average weight of 3.5 kg and age of 6 months have been involved in this trial. LSCs were isolated and cultured in vitro from the healthy eye of rabbits using a method modified by the authors. The abtained cells were then cultured for 14 days and transplanted to the collagen membrane, which was then examined using immunohistochemical analysis. RESULTS: The cells isolated from the biopsy were a mixture of fibroblast-type cells and cells with characteristics of LSC. They maintained high survivability, proliferativity, phenotype and stemness on the collagen carrier according to immunofluorescent study. CONCLUSIONS: Thus, the abstained tissue-engineered constructs could be used for further transplantation to the affected eye with limbal stem cell deficiency under experimental conditions.