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An important role for adenine, cholera toxin, hydrocortisone and triiodothyronine inthe proliferation, self-renewal and differentiation of limbal stem cells in vitro

Lookup NU author(s): Dr Min Yu, Dr Sanja Bojic, Dr Gustavo Figueiredo, Dr Julian De Havilland, Professor Anne Dickinson, Professor Francisco Figueiredo, Professor Majlinda Lako

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

The cornea is a self-renewing tissue located at the front of the eye. Its transparency is essential for allowing light to focusonto the retina for visual perception. The continuous renewal of corneal epithelium is supported by limbal stem cells(LSCs) which are located in the border region between conjunctiva and cornea known as the limbus. Ex vivo expansionof LSCs has been successfully applied in the last two decades to treat patients with limbal stem cell deficiency (LSCD).Various methods have been used for their expansion, yet the most widely used culture media contains a number of ingredientsderived from animal sources which may compromise the safety profile of human LSC transplantation. In this studywe sought to understand the role of these components namely adenine, cholera toxin, hydrocortisone and triiodothyroninewith the aim of re-defining a safe and GMP compatible minimal media for the ex vivo expansion of LSCs on humanamniotic membrane. Our data suggest that all four components play a critical role in maintaining LSC proliferation andpromoting LSC self-renewal. However removal of adenine and triiodothyronine had a more profound impact and led toLSC differentiation and loss of viability respectively, suggesting their essential role for ex vivo expansion of LSCs. Replacementof each of the components with GMP-grade reagents resulted in equal growth to non-GMP grade media, howeveran enhanced differentiation of LSCs was observed, suggesting that additional combinations of GMP grade reagentsneed to be tested to achieve similar or better level of LSC maintenance in the same manner as the traditional LSC media.


Publication metadata

Author(s): Yu M, Bojic S, Figueiredo GS, Rooney P, deHavilland J, Dickinson A, Figueiredo FC, Lako M

Publication type: Article

Publication status: Published

Journal: Experimental Eye Research

Year: 2016

Volume: 152

Pages: 113-122

Print publication date: 01/11/2016

Online publication date: 28/09/2016

Acceptance date: 27/09/2016

ISSN (print): 0014-4835

ISSN (electronic): 1096-0007

Publisher: Academic Press

URL: http://dx.doi.org/10.1016/j.exer.2016.09.008

DOI: 10.1016/j.exer.2016.09.008


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