Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering

The lack of donor corneal tissue or the immunological rejection remains a challenge for individuals with limbal stem cell deficiency (LSCD) who are treated with keratoplasty. Numerous lenticules which were extracted by small incision lenticule extraction (SMILE) appear to be useful materials for ker...

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Main Authors: Shuai Qin, Shuai Zheng, Bing Qi, Rui Guo, Guanghui Hou
Format: Article
Language:English
Published: Hindawi Limited 2019-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2019/4252514
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spelling doaj-e96173a8189c489581d50b8c3aae9ffe2020-11-25T01:49:59ZengHindawi LimitedStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/42525144252514Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue EngineeringShuai Qin0Shuai Zheng1Bing Qi2Rui Guo3Guanghui Hou4Department of Ophthalmology, Zhuhai Hospital Affiliated with Jinan University, Zhuhai People’s Hospital, Zhuhai, 519000 Guangdong, ChinaDepartment of Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, 515000 Guangdong, ChinaDepartment of Ophthalmology, Zhuhai Hospital Affiliated with Jinan University, Zhuhai People’s Hospital, Zhuhai, 519000 Guangdong, ChinaDepartment of Ophthalmology, Zhuhai Hospital Affiliated with Jinan University, Zhuhai People’s Hospital, Zhuhai, 519000 Guangdong, ChinaDepartment of Ophthalmology, Zhuhai Hospital Affiliated with Jinan University, Zhuhai People’s Hospital, Zhuhai, 519000 Guangdong, ChinaThe lack of donor corneal tissue or the immunological rejection remains a challenge for individuals with limbal stem cell deficiency (LSCD) who are treated with keratoplasty. Numerous lenticules which were extracted by small incision lenticule extraction (SMILE) appear to be useful materials for keratoplasty. In order to reduce the incidence of allograft rejection, lenticules would be decellularized. Lenticules which were treated with liquid nitrogen and nucleases had no cellular and nuclear materials remained. Human induced pluripotent stem cells (iPSCs) can be generated from the patient who requires keratoplasty, offering an autologous alternative and eliminating the risk of graft rejection. We found that BMP-4, RA, N-2 supplement, hEGF, B27, decellularized human stromal lenticules, conditioned medium, or induction medium promoted the differentiation of human iPSCs with high purity. The results showed that human iPSCs cultured for 4 days in differentiation medium A, 14 days in condition medium, and 1 week in induction medium on decellularized human stromal lenticules developed markedly higher expression of the markers P63, CK3, and CK12 than did those in the other methods. The level of gene expression of the epithelial and pluripotency markers and analysis by scanning electron microscopy and immunohistochemistry also showed successful differentiation. After inducing differentiation in vitro, corneal epithelial-like cells were induced. In the study, we investigated the possibility of a new resource for corneal tissue engineering.http://dx.doi.org/10.1155/2019/4252514
collection DOAJ
language English
format Article
sources DOAJ
author Shuai Qin
Shuai Zheng
Bing Qi
Rui Guo
Guanghui Hou
spellingShingle Shuai Qin
Shuai Zheng
Bing Qi
Rui Guo
Guanghui Hou
Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering
Stem Cells International
author_facet Shuai Qin
Shuai Zheng
Bing Qi
Rui Guo
Guanghui Hou
author_sort Shuai Qin
title Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering
title_short Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering
title_full Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering
title_fullStr Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering
title_full_unstemmed Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering
title_sort decellularized human stromal lenticules combine with corneal epithelial-like cells: a new resource for corneal tissue engineering
publisher Hindawi Limited
series Stem Cells International
issn 1687-966X
1687-9678
publishDate 2019-01-01
description The lack of donor corneal tissue or the immunological rejection remains a challenge for individuals with limbal stem cell deficiency (LSCD) who are treated with keratoplasty. Numerous lenticules which were extracted by small incision lenticule extraction (SMILE) appear to be useful materials for keratoplasty. In order to reduce the incidence of allograft rejection, lenticules would be decellularized. Lenticules which were treated with liquid nitrogen and nucleases had no cellular and nuclear materials remained. Human induced pluripotent stem cells (iPSCs) can be generated from the patient who requires keratoplasty, offering an autologous alternative and eliminating the risk of graft rejection. We found that BMP-4, RA, N-2 supplement, hEGF, B27, decellularized human stromal lenticules, conditioned medium, or induction medium promoted the differentiation of human iPSCs with high purity. The results showed that human iPSCs cultured for 4 days in differentiation medium A, 14 days in condition medium, and 1 week in induction medium on decellularized human stromal lenticules developed markedly higher expression of the markers P63, CK3, and CK12 than did those in the other methods. The level of gene expression of the epithelial and pluripotency markers and analysis by scanning electron microscopy and immunohistochemistry also showed successful differentiation. After inducing differentiation in vitro, corneal epithelial-like cells were induced. In the study, we investigated the possibility of a new resource for corneal tissue engineering.
url http://dx.doi.org/10.1155/2019/4252514
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