Five-Year Follow-up of First 11 Patients Undergoing Injection of Cultured Corneal Endothelial Cells for Corneal Endothelial Failure

Numa, Kohsaku; Imai, Kojiro; Ueno, Masaki; Kitazawa, Koji; Tanaka, Hiroshi; Bush, John; Teramukai, Satoshi; Okumura, Naoki; Koizumi, Noriko; Hamuro, Junji; Sotozono, Chie; Kinoshita, Shigeru

doi:10.1016/j.ophtha.2020.09.002

Cited by 86 publications

(86 citation statements)

References 44 publications

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“…A prospective observational study confirmed that at 5 years after surgery, CE function was restored in 10 of the 11 eyes, the mean central corneal ECD of which was 1257 ± 467 cells/mm 2 . This follow-up study confirmed the safety and efficacy of the developed cell-injection therapy method for treating bullous keratopathy patients [ 25 ].…”

Section: Transplantation Of Bioengineered Corneal Endotheliumsupporting

confidence: 65%

“…This review shows that the introduction of a robust dual-media culture system for the ex vivo expansion of CECs, as well as intensive research into selected signaling pathway inhibitors, such as Rho-associated kinase inhibitors, has enabled the preparation of GMP-compliant CE grafts for first-in-human clinical trials with promising outcomes [ 24 , 25 ]. According to the study, approximately 300 patients (eyes) can be treated using T-E grafts derived from one (younger) donor cadaveric cornea, using culture conditions developed by authors [ 26 , 27 ].…”

Section: Introductionmentioning

confidence: 99%

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Ex vivo expansion and characterization of human corneal endothelium for transplantation: a review

2021

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The corneal endothelium plays a key role in maintaining corneal transparency. Its dysfunction is currently treated with penetrating or lamellar keratoplasty. Advanced cell therapy methods seek to address the persistent global deficiency of donor corneas by enabling the renewal of the endothelial monolayer with tissue-engineered grafts. This review provides an overview of recently published literature on the preparation of endothelial grafts for transplantation derived from cadaveric corneas that have developed over the last decade (2010–2021). Factors such as the most suitable donor parameters, culture substrates and media, endothelial graft storage conditions, and transplantation methods are discussed. Despite efforts to utilize alternative cellular sources, such as induced pluripotent cells, cadaveric corneas appear to be the best source of cells for graft preparation to date. However, native endothelial cells have a limited natural proliferative capacity, and they often undergo rapid phenotype changes in ex vivo culture. This is the main reason why no culture protocol for a clinical-grade endothelial graft prepared from cadaveric corneas has been standardized so far. Currently, the most established ex vivo culture protocol involves the peel-and-digest method of cell isolation and cell culture by the dual media method, including the repeated alternation of high and low mitogenic conditions. Culture media are enriched by additional substances, such as signaling pathway (Rho-associated protein kinase, TGF-β, etc.) inhibitors, to stimulate proliferation and inhibit unwanted morphological changes, particularly the endothelial-to-mesenchymal transition. To date, this promising approach has led to the development of endothelial grafts for the first in-human clinical trial in Japan. In addition to the lack of a standard culture protocol, endothelial-specific markers are still missing to confirm the endothelial phenotype in a graft ready for clinical use. Because the corneal endothelium appears to comprise phenotypically heterogeneous populations of cells, the genomic and proteomic expression of recently proposed endothelial-specific markers, such as Cadherin-2, CD166, or SLC4A11, must be confirmed by additional studies. The preparation of endothelial grafts is still challenging today, but advances in tissue engineering and surgery over the past decade hold promise for the successful treatment of endothelial dysfunctions in more patients worldwide.

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Section: Transplantation Of Bioengineered Corneal Endotheliumsupporting

confidence: 65%

Section: Introductionmentioning

confidence: 99%

Ex vivo expansion and characterization of human corneal endothelium for transplantation: a review

2021

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“…Discussing the implementation of the suggested here technique into current industry standards, we can make simple calculations: during 7–9 weeks of co-cultivation (about 10 passages) at the passaging diluting of 4 to 6 fold the whole procedure can give up to ∼5 10 = ∼10 7 fold multiplication of the initial cell number (let say 10 5 cells), resulting in 10 12 cells, which is comparable to the amount of the cells in human body. Current protocols of human EC therapy requires from 1.5 × 10 6 ( Numa et al, 2020 ) to 2 × 10 9 cells (calculation based on human/mouse weight ratio) ( Poulos et al, 2015 ). Since it could be difficult to produce such amount of cells in standard laboratory 2D plates (2 × 10 9 cells would occupy about 2 m 2 ), the industrial 3D cell culture systems should be used, and appropriate bioprocess screening technologies applied ( Abraham et al, 2018 ).…”

Section: Discussionmentioning

confidence: 99%

Co-growth of Stem Cells With Target Tissue Culture as an Easy and Effective Method of Directed Differentiation

Kovina

Dyuzheva

Krasheninnikov

et al. 2021

Front. Bioeng. Biotechnol.

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The long-term co-culture of mouse embryonic stem cells (mESC) with rat endothelial cells (EC) was tested for contact differentiation into the endothelial lineage. Serial passaging of rat ECs mixed with mESC in ratio 10:1 resulted in the emergence of a homogeneous cell population expressing mouse endothelial surface markers CD102, CD29, CD31. Rat endothelial surface marker RECA-1 completely disappeared from the co-cultured population after 2 months of weekly passaging. Co-incubation of mESC with rat ECs without cell-to-cell contact did not result in the conversion of mESC into ECs. After co-cultivation of adult mesenchymal stem cells from human endometrium (eMSC) with pre-hepatocyte-like cells of human hepatocarcinoma Huh7 the resulting co-culture expressed mature liver markers (oval cell antigen and cytokeratin 7), none of which were expressed by any of co-cultivated cultures, thus proving that even an immature (proliferating) pre-hepatocyte-like line can induce hepatic differentiation of stem cells. In conclusion, we have developed conditions where long-term co-proliferation of embryonic or adult SC with fully or partially differentiated cells results in stem cell progeny expressing markers of target tissue. In the case of endothelial differentiation, the template population quickly disappeared from the resulted culture and the pure endothelial population of stem cell progeny emerged. This approach demonstrates the expected fate of stem cells during various in vivo SC-therapies and also might be used as an effective in vitro differentiation method to develop the pure endothelium and, potentially, other tissue types of desirable genetic background.

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“…Promising results were observed up to 5-year follow-up, proving HCEnC therapy as a potential alternative to corneal transplantation for managing endothelial diseases. [38][39][40] Similarly, the Singapore group has demonstrated that the corneal transparency in rabbit eyes with corneal endothelial dysfunction could be restored using a regulatory-compliant cell culture system, with the cells delivered either as tissue-engineered endothelial keratoplasty (TE-EK) or as cell injection therapy. 12 We have recently initiated a first-in-human trial of HCEnC therapy for corneal endothelial disease in Singapore, having received Institutional Review Board (IRB) approval in 2016.…”

Section: Overview Of Corneal Endothelial Diseases and Potential Impact Of Cell Therapymentioning

confidence: 99%

Translational and Regulatory Challenges of Corneal Endothelial Cell Therapy: A Global Perspective

Ting

Peh

Adnan

2022

Tissue Engineering Part B: Reviews

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Cell therapies are emerging as a unique class of clinical therapeutics in medicine. In 2015, Holoclar (ex vivo expanded autologous human corneal epithelial cells containing stem cells) gained the regulatory approval for treating limbal stem cell deficiency after chemical eye burn. This has set a precedent in ophthalmology and in medicine, reinforcing the therapeutic promise of cell therapy. However, to generalize and commercialize cell therapies on a global scale, stringent translational and regulatory requirements need to be fulfilled at both local and international levels. Over the past decade, the Singapore group has taken significant steps in developing human corneal endothelial cell (HCEnC) therapy for treating corneal endothelial diseases, which are currently the leading indication for corneal transplantation in many countries. Successful development of HCEnC therapy may serve as a novel solution to the current global shortage of donor corneas. Based on the experience in Singapore, this review aims to provide a global perspective on the translational and regulatory challenges for bench-to-bedside translation of cell therapy. Specifically, we discussed about the characterization of the critical quality attributes (CQA), the challenges that can affect the CQA, and the variations in the regulatory framework embedded within different regions, including Singapore, Europe, and the United States.

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Five-Year Follow-up of First 11 Patients Undergoing Injection of Cultured Corneal Endothelial Cells for Corneal Endothelial Failure

Cited by 86 publications

References 44 publications

Ex vivo expansion and characterization of human corneal endothelium for transplantation: a review

Ex vivo expansion and characterization of human corneal endothelium for transplantation: a review

Co-growth of Stem Cells With Target Tissue Culture as an Easy and Effective Method of Directed Differentiation

Translational and Regulatory Challenges of Corneal Endothelial Cell Therapy: A Global Perspective

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