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Original Article
124 (
6
); 655-664
doi:
10.25259/IJMR_20061246_655

Cultivation of human corneal limbal stem cells in Mebiol gel® - A thermo-reversible gelation polymer

Vision Research Foundation, Sankara Nethralaya, Chennai
Advanced Research Center for Science & Engineering, Waseda University, Tokyo, Japan
Department of Surgery, Yamanashi University- School of Medicine, Yamanashi, Japan

Present address: Nichi-In Centre for Regenerative Medicine, Vijaya Health Centre Premises NSK Road, Chennai 600094, India

Reprint requests: Dr H.N. Madhavan, Vision Research Foundation, Sankara Nethralaya 18, College Road, Chennai 600006, India e-mail: drhnm@snmail.org

Licence
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background & objectives:

Cultivated limbal stem cell transplantation is being used as a current treatment modality for limbal stem cell deficiency. However, use of allogenic biological material as substrate is associated with risks of transmission of certain diseases and allograft rejection. Therefore development of non-toxic biodegradable synthetic polymers is important. We undertook this study to evaluate the use of a synthetic polymer Mebiol gel as a substrate for the growth of limbal phenotype cells and cornea phenotype cells from limbal explants.

Methods:

Human cadaveric limbal explants cells were cultivated on Mebiol gel. The proliferative capacity of cultivated cells was analyzed with thymidine incorporation studies. Immunostaining for presumed limbal stem cell association markers and cornea differentiation markers was performed and confirmed with reverse transcription (RT-PCR).

Results:

The limbal explants underwent proliferation in vitro. The cultivated cells expressed the presumed limbal stem cell association markers (ABCG2 and p63), the transient amplifying cell markers (connexin 43, integrin a9) and the cornea differentiation marker (K3). RT PCR confirmed the immunohistochemical data.

Interpretation & conclusion:

Our findings showed that the synthetic polymer Mebiol gel was able to support limbal explant proliferation. The cultured cells expressed presumed limbal stem cell association markers, transient amplifying cells and cornea phenotype markers. Mebiol Gel can be used as a scaffold for growing limbal explants.

Keywords

Limbal explants
limbal stem cell transplation
Mebiol gel®

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