Long-Term Observation and Sequencing Analysis of SKPs-Derived Corneal Endothelial Cell-Like Cells for Treating Corneal Endothelial Dysfunction

Corneal endothelial dysfunction is a principal cause of visual deficiency. Corneal transplantation is the most effective treatment for corneal endothelial dysfunction. However, a severe shortage of available donor corneas or human corneal endothelial cells (HCECs) remains a global challenge. Previously, we acquired corneal endothelial cell-like cells (CEC-like cells) derived from human skin-derived precursors (SKPs). CEC-like cells were injected into rabbit and monkey corneal endothelial dysfunction models and exerted excellent therapeutic effect. In this study, we prolonged the clinical observation in the monkey experiment for 2 years. Polymerase chain reaction (PCR) and DNA sequencing were carried out to confirm the existence of CEC-like cells. Histological examinations were carried out to show the corneal morphology. Further transcriptome sequencing was also carried out on HCEC, CEC-like cells before transplantation and after transplantation. We found that the monkeys cornea remained transparent and normal thickness. The total endothelial cell density decreased gradually, but tended to be stable and remained in a normal range during 2-year observation. The CEC-like cells persist during observation and could adapt to the microenvironment after transplantation. The gene expression pattern of CEC-like cells was similar to HCEC and changed slightly after transplantation. In conclusion, this study presented a brand-new insight into CEC-like cells and further provided a promising prospect of cell-based therapy for corneal endothelial dysfunction. The renewable cell source, novel derivation method and simple treatment strategy may be clinically applied in regenerative medicine in the future.

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Long-term observation and sequencing analysis of SKPs-derived corneal endothelial cell-like cells for treating corneal endothelial dysfunction

10.21203/rs.3.rs-36046/v1 ◽

2020 ◽

Author(s):

Lin Shen ◽

Peng Sun ◽

Liqun Du ◽

Jing Zhu ◽

Chengqun Ju ◽

...

Keyword(s):

Endothelial Cell ◽

Endothelial Dysfunction ◽

Corneal Transplantation ◽

Gene Expression Pattern ◽

Corneal Endothelial Cell ◽

Sequencing Analysis ◽

Global Challenge ◽

Cell Based Therapy ◽

Corneal Endothelial Dysfunction ◽

Long Term Observation

Abstract Background Corneal endothelial dysfunction is a principal cause of visual deficiency. Corneal transplantation is the most effective treatment for corneal endothelial dysfunction. However, a severe shortage of available donor corneas remains a global challenge. Previously, we acquired corneal endothelial cell-like cells (CEC-like cells) derived from skin-derived precursors (SKPs). CEC-like cells exerted excellent therapeutic effect in rabbit and monkey corneal endothelial dysfunction models. Method: We prolonged the observation in the monkey experiment for 2 years. Further transcriptome sequencing was also carried out on HCEC, CEC-like cells before transplantation and after transplantation. Results We confirmed that the monkey cornea remained transparent and the CEC-like cells could adapt to the microenvironment after transplantation. The gene expression pattern of CEC-like cells was similar to HCEC and changed slightly after transplantation. Conclusions This study presented a brand-new insight into CEC-like cells and further provided a promising prospect of cell-based therapy for corneal endothelial dysfunction. The renewable cell source, novel derivation method and simple treatment strategy may be clinically applied in regenerative medicine in the future.

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Long-term observation and sequencing analysis of SKPs-derived corneal endothelial cell-like cells for treating corneal endothelial dysfunction

10.21203/rs.3.rs-36046/v2 ◽

2020 ◽

Author(s):

Lin Shen ◽

Peng Sun ◽

Liqun Du ◽

Jing Zhu ◽

Chengqun Ju ◽

...

Keyword(s):

Endothelial Cell ◽

Endothelial Dysfunction ◽

Therapeutic Effect ◽

Gene Expression Pattern ◽

Corneal Endothelial Cell ◽

Sequencing Analysis ◽

Cell Based Therapy ◽

Corneal Endothelial Dysfunction ◽

Long Term Observation

Abstract Background Corneal endothelial dysfunction is a principal cause of visual deficiency. Corneal transplantation is the most effective treatment for corneal endothelial dysfunction. However, a severe shortage of available donor corneas or human corneal endothelial cells (HCEC) remains a global challenge. Previously, we acquired corneal endothelial cell-like cells (CEC-like cells) derived from human skin-derived precursors (SKPs). CEC-like cells were injected into rabbit and monkey corneal endothelial dysfunction models and exerted excellent therapeutic effect. Method We prolonged the clinical observation in the monkey experiment for 2 years. PCR and DNA sequencing were carried out to confirm the existence of CEC-like cells. Histological examinations were carried out to show the corneal morphology. Further transcriptome sequencing was also carried out on HCEC, CEC-like cells before transplantation and after transplantation. Results The monkeys cornea remained transparent and normal thickness. The total endothelial cell density decreased gradually, but tended to be stable and remained in a normal range during 2-year observation. The CEC-like cells persist during observation and could adapt to the microenvironment after transplantation. The gene expression pattern of CEC-like cells was similar to HCEC and changed slightly after transplantation. Conclusions CEC-like cells could adapt to the microenvironment and had a good therapeutic effect after being transplanted into the monkey endothelial dysfunction model during long-term observation. This study provided a promising prospect of cell-based therapy for corneal endothelial dysfunction and may be clinically applied in regenerative medicine in the future.

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Review: corneal endothelial cell derivation methods from ES/iPS cells

Inflammation and Regeneration ◽

10.1186/s41232-019-0108-y ◽

2019 ◽

Vol 39 (1) ◽

Cited By ~ 5

Author(s):

Shin Hatou ◽

Shigeto Shimmura

Keyword(s):

Endothelial Cell ◽

Conditioned Medium ◽

Corneal Transplantation ◽

Ips Cells ◽

Lens Epithelial Cell ◽

Corneal Endothelial Cell ◽

Bullous Keratopathy ◽

Corneal Endothelial Dysfunction ◽

Fetal Bovine ◽

Proof Of Efficacy

Abstract Globally, approximately 12.7 million people are awaiting a transplantation, while only 185,000 cases of corneal transplantation are performed in a year. Corneal endothelial dysfunction (bullous keratopathy) due to Fuchs’ corneal endothelial dystrophy, or insults associated with intraocular surgeries, shared half of all indications for corneal transplantation. Regenerative therapy for corneal endothelium independent of eye bank eyes has great importance to solve the large supply-demand mismatching in corneal transplantation and reduce the number of worldwide corneal blindness. If corneal endothelial cells could be derived from ES or iPS cells, these stem cells would be the ideal cell source for cell therapy treatment of bullous keratopathy. Four representative corneal endothelial cell derivation methods were reviewed. Components in earlier methods included lens epithelial cell-conditioned medium or fetal bovine serum, but the methods have been improved and materials have been chemically more defined over the years. Conditioned medium or serum is replaced to recombinant proteins and small molecule compounds. These improvements enabled to open the corneal endothelial developmental mechanisms, in which epithelial-mesenchymal and mesenchymal-endothelial transition by TGF beta, BMP, and Wnt signaling have important roles. The protocols are gradually approaching clinical application; however, proof of efficacy and safety of the cells by adequate animal models are the challenges for the future.

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Corneal endothelial cell dysfunction: etiologies and management

Therapeutic Advances in Ophthalmology ◽

10.1177/2515841418815802 ◽

2018 ◽

Vol 10 ◽

pp. 251584141881580 ◽

Cited By ~ 13

Author(s):

Sepehr Feizi

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Endothelial Dysfunction ◽

Corneal Endothelial Cell ◽

Endothelial Cell Dysfunction ◽

Endothelial Keratoplasty ◽

Corneal Endothelial Cells ◽

Cell Dysfunction ◽

Corneal Endothelial Dysfunction

A transparent cornea is essential for the formation of a clear image on the retina. The human cornea is arranged into well-organized layers, and each layer plays a significant role in maintaining the transparency and viability of the tissue. The endothelium has both barrier and pump functions, which are important for the maintenance of corneal clarity. Many etiologies, including Fuchs’ endothelial corneal dystrophy, surgical trauma, and congenital hereditary endothelial dystrophy, lead to endothelial cell dysfunction. The main treatment for corneal decompensation is replacement of the abnormal corneal layers with normal donor tissue. Nowadays, the trend is to perform selective endothelial keratoplasty, including Descemet stripping automated endothelial keratoplasty and Descemet’s membrane endothelial keratoplasty, to manage corneal endothelial dysfunction. This selective approach has several advantages over penetrating keratoplasty, including rapid recovery of visual acuity, less likelihood of graft rejection, and better patient satisfaction. However, the global limitation in the supply of donor corneas is becoming an increasing challenge, necessitating alternatives to reduce this demand. Consequently, in vitro expansion of human corneal endothelial cells is evolving as a sustainable choice. This method is intended to prepare corneal endothelial cells in vitro that can be transferred to the eye. Herein, we describe the etiologies and manifestations of human corneal endothelial cell dysfunction. We also summarize the available options for as well as recent developments in the management of corneal endothelial dysfunction.

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Therapy of corneal endothelial dysfunction with corneal endothelial cell-like cells derived from skin-derived precursors

Scientific Reports ◽

10.1038/s41598-017-13787-1 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 14

Author(s):

Lin Shen ◽

Peng Sun ◽

Canwei Zhang ◽

Le Yang ◽

Liqun Du ◽

...

Keyword(s):

Endothelial Cell ◽

Endothelial Dysfunction ◽

Corneal Endothelial Cell ◽

Corneal Endothelial Dysfunction

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Cases With Long-Term High Corneal Endothelial Cell Density Maintained After Corneal Transplantation

Cornea ◽

10.1097/ico.0000000000002386 ◽

2020 ◽

Vol Publish Ahead of Print ◽

Cited By ~ 1

Author(s):

Kanae Kayukawa ◽

Koji Kitazawa ◽

Koichi Wakimasu ◽

Sanjay V. Patel ◽

John Bush ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Density ◽

Corneal Transplantation ◽

Corneal Endothelial Cell ◽

Endothelial Cell Density ◽

Corneal Endothelial Cell Density

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Generation and Feasibility Assessment of a New Vehicle for Cell-Based Therapy for Treating Corneal Endothelial Dysfunction

PLoS ONE ◽

10.1371/journal.pone.0158427 ◽

2016 ◽

Vol 11 (6) ◽

pp. e0158427 ◽

Cited By ~ 13

Author(s):

Naoki Okumura ◽

Kazuya Kakutani ◽

Ryota Inoue ◽

Daiki Matsumoto ◽

Tomoki Shimada ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Feasibility Assessment ◽

Cell Based Therapy ◽

Corneal Endothelial Dysfunction

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Corneal Endothelial Cell Density and Morphology in Healthy Egyptian Eyes

Journal of Ophthalmology ◽

10.1155/2019/6370241 ◽

2019 ◽

Vol 2019 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Marwa Mahmoud Abdellah ◽

Hatem Gamal Ammar ◽

Mohamed Anbar ◽

Engy Mohammed Mostafa ◽

Mahmoud Mohamed Farouk ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Density ◽

Corneal Thickness ◽

Corneal Transplantation ◽

Corneal Endothelial Cell ◽

Endothelial Cell Density ◽

Specular Microscopy ◽

Cell Area ◽

Corneal Endothelial Cell Density ◽

The Mean

Purpose. To evaluate the corneal endothelial cell density and morphology in normal Egyptian eyes. Methods. In total, 568 healthy eyes of 568 Egyptian volunteers aged 20 to 85 years were examined using noncontact specular microscopy for the central corneal thickness (CCT), mean endothelial cell density (MCD), coefficient of variation (CV) in cell area, mean cell area (MCA), and hexagonal cell (Hex) percentage. Variables were compared between sexes and between different age groups. Results. The mean CCT, MCD, and MCA were 514.45 ± 43.04 μm, 2647.50 ± 382.62 cells/mm2, and 390.59 ± 149.94 μm2, respectively. MCD and MCA showed no significant differences between men and women (P=0.171 and 0.099, respectively), whereas CV (%) and Hex (%) showed significant differences (P=0.024 and 0.015, respectively). CCT (P=0.007, r = −0.113) and MCD (P<0.001, r = −0.357) exhibited a significant negative correlation with age, whereas CV (%) (P<0.001, r = 0.341) and MCA (P=0.008, r = 0.111) exhibited a significant positive correlation. The mean rate of endothelial cell loss from 20 to 85 years of age was 0.3% per year. Conclusions. Our results provide normative data for the corneal endothelium in healthy Egyptian eyes, thus increasing the knowledge base for corneal endothelial cell parameters in healthy Egyptian eyes. Furthermore, our findings can be used as baseline values for comparisons between Egyptian and other populations and for studies of the endothelial cell reserve and capacity for intraocular surgery and corneal transplantation.

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