Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Aug 26, 2017; 9(8): 127-132
Published online Aug 26, 2017. doi: 10.4252/wjsc.v9.i8.127
Transplanting embryonic stem cells onto damaged human corneal endothelium
Charles Hanson, Arsaell Arnarsson, Thorir Hardarson, Ann Lindgård, Mandana Daneshvarnaeini, Catarina Ellerström, Anita Bruun, Ulf Stenevi
Charles Hanson, Unit of Reproductive Medicine, Sahlgrenska University Hospital, 41345 Gothenburg, Sweden
Arsaell Arnarsson, Neuroscience Laboratory, University of Akureyri, 600 Akureyri, Iceland
Thorir Hardarson, Fertility Centre Scandinavia, Carlanderska Hospital, 40229 Gothenburg, Sweden
Ann Lindgård, Mandana Daneshvarnaeini, Ulf Stenevi, Department of Ophthalmology, Gothenburg University, 43180 Mölndal, Sweden
Catarina Ellerström, Takara Bio Europe AB, Arvid Wallgrens Backe 20, 41346 Gothenburg, Sweden
Anita Bruun, Department of Ophthalmology, Lund University Hospital, 22121 Lund, Sweden
Author contributions: Hanson C, Arnarsson A, Hardarson T and Stenevi U designed the research; Ellerström C was responsible for the undifferentiated pluripotent human embryonic stem cells; Stenevi U was responsible for preparation of the human corneas; Hanson C, Arnarsson A, Lindgård A, Daneshvarnaeini M and Bruun A performed the research; all authors were involved in the manuscript preparation.
Supported by De Blindas Vänner, Gothenburg, and Greta Bergs Foundation, Lerum (to Charles Hanson); University of Akureyri Research Fund, the KEA Fund, and the Icelandic Council on Ageing (to Arsaell Arnarsson); and Gothenburg Medical Society, the Medical Faculty of the University of Gothenburg and the Herman Svensson Foundation (to Ulf Stenevi).
Institutional review board statement: This study has been reviewed and approved by the Regional Ethics Committee in Gothenburg and Malmö (approval number 067-04 and M155-14).
Institutional animal care and use committee statement: No animals were used in this study.
Conflict-of-interest statement: Catarina Ellerström is an employee of Takara Bio Europe AB. To the best of our knowledge, no conflict of interest exists for the other authors.
Data sharing statement: Dataset available from the corresponding author (charles.hanson@obgyn.gu.se). Participants gave informed consent for anonymized data sharing. All data are anonymized and risk of identification is low.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Charles Hanson, PhD, Associate Professor of Reproductive Genetics, Unit of Reproductive Medicine, Sahlgrenska University Hospital, Blå stråket 6, 41345 Gothenburg, Sweden. charles.hanson@obgyn.gu.se
Telephone: +46-31-3423572
Received: January 24, 2017
Peer-review started: February 8, 2017
First decision: April 17, 2017
Revised: June 30, 2017
Accepted: July 14, 2017
Article in press: July 17, 2017
Published online: August 26, 2017
Processing time: 211 Days and 3.1 Hours
Abstract
AIM

To investigate whether human embryonic stem cells (hESCs) could be made to attach, grow and differentiate on a human Descemet’s membrane (DM).

METHODS

Spontaneously differentiated hESCs were transferred onto a human corneal button with the endothelial layer removed using ocular sticks. The cells were cultured on a DM for up to 15 d. The genetically engineered hESC line expressed green fluorescent protein, which facilitated identification during the culture experiments, tissue preparation, and analysis. To detect any differentiation into human corneal endothelial-like cells, we analysed the transplanted cells by immunohistochemistry using specific antibodies.

RESULTS

We found transplanted cells form a single layer of cells with a hexagonal shape in the periphery of the DM. The majority of the cells were negative for octamer-binding transcription factor 4 but positive for paired box 6 protein, sodium potassium adenosine triphosphatase (NaKATPase), and Zona Occludens protein 1. In four of the 18 trials, the transplanted cells were found to express CK3, which indicates that the stem cells differentiated into corneal epithelial cells in these cases.

CONCLUSION

It is possible to get cells originating from hESCs to become established on a human DM, where they grow and differentiate into corneal endothelial-like cells in vitro.

Keywords: Embryonic stem cells; Cornea; Descemet’s membrane; Endothelium; Immunohistochemistry

Core tip: This is the first report on the interaction between human embryonic stem cells (hESCs) and the inner parts of the human cornea. When hESCs were transplanted onto Descemet’s membrane (DM) in vitro we found that they were able to attach to and grow on DM in a single cell layer. Furthermore, the stem cells changed their morphology from small round cells to flat hexagonal cells. The transplanted hESCs also started to express the proteins paired box 6, Zona Occludens protein 1 and sodium potassium adenosine triphosphatase (NaKATPase), which also are expressed in human corneal endothelial cells.