Corneal Endothelial Regeneration Using Mesenchymal Stem Cells Derived from Human Umbilical Cord

Yamashita, Kazuya; Inagaki, Emi; Hatou, Shin; Higa, Kazunari; Ogawa, Akiko; Miyashita, Hideyuki; Ohta, Michio; Shimmura, Shigeto

doi:10.1089/scd.2017.0297

Cited by 54 publications

(50 citation statements)

References 43 publications

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“…Although the gene expression was higher in CEC-derived from DPSC, its value was lower than the expression of human CEC, used as control. This is in line with a previous study which presented lower levels of gene expression of CEC-derived from ESC compared to human CEC or CEC cell line [30,45].…”

Section: Discussionsupporting

confidence: 93%

A new alternative for corneal endothelial regeneration using autologous dental pulp stem cells

Bosch

Salero²,

Núñez-Toldrà

et al. 2020

Preprint

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Background Failure of the corneal endothelium cell (CEC) monolayer is the main cause leading to corneal transplantation. Autologous cell-based therapies are required to reconstruct in vitro the cell monolayer. For this purpose, we propose the use of dental pulp stem cells isolated from the third molars to form CEC monolayer. We hypothesize that by using dental pulp stem cells (DPSC) that share an embryological origin with CEC, as they both arise from the neural crest, may allow a direct differentiation process avoiding the use of reprogramming techniques, such as induced pluripotent stem cells (iPSC). Methods In this work, we report a two-step differentiation protocol, where dental pulp stem cells are derived into neural crest stem cells and, then, into CEC. Results Initially, we compared the efficiency of direct differentiation of DPSC with the differentiation of iPSC to express NCSC related genes in adhesion culture, showing significantly higher levels of early stage marker AP2 for the DPSC compared to iPSC. To provide better environment for NCSC gene expression, suspension method was performed, which induced the formation of neurospheres. The results showed neurosphere formation after few days, obtaining the peak of NCSC marker expression after 4 days, showing overexpression of AP2, p75 and CHD7 markers, confirming the formation of NCSC like cells. Furthermore, pluripotent markers Oct4, Nanog and Sox2 were as well upregulated in suspension culture. Neurospheres were then directly cultured in CEC conditioned medium for the second differentiation, showing the conversion of DPSC into polygonal like cells expressing higher levels of ZO-1, ATP1A1, COL4A2 and COL8A1 markers, providing proof of the successful conversion into CEC. Conclusions Therefore, our findings demonstrate that patient-derived dental pulp stem cells represent an autologous cell source for corneal endothelial regeneration that avoids actual transplantation limitations as well as reprogramming techniques.

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Section: Discussionsupporting

confidence: 93%

A new alternative for corneal endothelial regeneration using autologous dental pulp stem cells

Bosch

Salero²,

Núñez-Toldrà

et al. 2020

Preprint

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“…The worldwide shortage of donor corneal tissue for the treatment of corneal endothelial dysfunction necessitates the development of viable alternatives to the paradigm of one donor cornea being used for only one recipient [10][11][12] . The in vitro generation of stem cell-derived corneal endothelial-like cells 13 , immortalized CEnC lines 14,15 and expansion of primary CEnC from cadaveric donor corneal tissue 10,16,17 have challenged the one donor-one recipient paradigm of corneal transplantation. Nevertheless, in vitro culture poses its own challenges, including unwanted changes in cell phenotype (e.g., endothelial to fibroblastic) and progression towards replicative senescence that limits cell numbers 11,18 .…”

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confidence: 99%

Phenotypic and functional characterization of corneal endothelial cells during in vitro expansion

Frausto¹,

Swamy²,

Peh

et al. 2020

Sci Rep

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the advent of cell culture-based methods for the establishment and expansion of human corneal endothelial cells (CEnC) has provided a source of transplantable corneal endothelium, with a significant potential to challenge the one donor-one recipient paradigm. However, concerns over cell identity remain, and a comprehensive characterization of the cultured cenc across serial passages has not been performed. to this end, we compared two established cenc culture methods by assessing the transcriptomic changes that occur during in vitro expansion. In confluent monolayers, low mitogenic culture conditions preserved corneal endothelial cell state identity better than culture in high mitogenic conditions. expansion by continuous passaging induced replicative cell senescence. transcriptomic analysis of the senescent phenotype identified a cell senescence signature distinct for CEnC. We identified activation of both classic and new cell signaling pathways that may be targeted to prevent senescence, a significant barrier to realizing the potential clinical utility of in vitro expansion. The corneal endothelium is a neuroectoderm-derived tissue that is located on the posterior surface of the cornea and is a semipermeable monolayer of mitotically inactive (i.e., quiescent) cells. A critical functional property of the corneal endothelium is to maintain the corneal stroma in a relatively dehydrated state. This process ensures that the collagen fibers of the stroma retain an ultrastructural organization essential for corneal transparency. The pump-leak hypothesis is believed to best explain the role that the endothelium plays in maintaining a relatively dehydrated stroma. Passive movement of water from the aqueous humor to the stroma (i.e., leak) and active transport of solutes (i.e., pump) in the opposite direction are regulated by the corneal endothelial cell (CEnC) barrier formation (i.e., tight junctions, cell adhesion) and expression of solute transporters (e.g., Na/K ATPases, SLC4A11) 1. Mutations in genes encoding solute transporters (e.g., SLC4A11) or other essential endothelial cell functions (e.g., ZEB1, TCF4) lead to stromal edema and loss of corneal clarity 2-5. In general, loss of barrier integrity, dysfunction of solute transporters or a significant decrease in CEnC density leads to the loss of corneal clarity that necessitates corneal transplantation. Endothelial cell failure constitutes the primary indication for corneal transplantation both worldwide and in the U.S., serving as the indication for 55% of all keratoplasty procedures performed in the US in 2018 6. While endothelial keratoplasty represents a significant advance in the surgical management of corneal endothelial dysfunction, a variety of associated intraoperative and postoperative complications, in addition to the low rates of donor corneal recovery and the lack of adequately trained surgeons in the majority of countries, have significantly limited the impact of endothelial keratoplasty on visual impairment due to corneal endothelial dysfunction worl...

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“…A certain number of studies have been done to isolate and characterize multipotent stem cells from different tissues in order to use their great potential in regenerative medicine. Bone marrow-derived mesenchymal stem cells [83], human umbilical cord mesenchymal stem cells [84], postnatal periodontal ligament [85], and limbal stem cells [86] are recently studied stem cells sources in corneal wound healing and regeneration. Saghizadeh et al [30] have recently reviewed all major stem cell usage in corneal wound healing.…”

Section: Opportunities and Challenges In Regenerative Ophthalmologymentioning

confidence: 99%

Corneal Stem Cells as a Source of Regenerative Cell-Based Therapy

Nurković

Vojinović

Dolićanin

2020

Stem Cells International

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In the past few years, intensive research has focused on corneal stem cells as an unlimited source for cell-based therapy in regenerative ophthalmology. Today, it is known that the cornea has at least two types of stem cells: limbal epithelial stem cells (LESCs) and corneal stromal stem cells (CSSCs). LESCs are used for regeneration of corneal surface, while CSSCs are used for regeneration of corneal stroma. Until now, various approaches and methods for isolation of LESCs and CSSCs and their successful transplantation have been described and tested in several preclinical studies and clinical trials. This review describes in detail phenotypic characteristics of LESCs and CSSCs and discusses their therapeutic potential in corneal regeneration. Since efficient and safe corneal stem cell-based therapy is still a challenging issue that requires continuous cooperation between researchers, clinicians, and patients, this review addresses the important limitations and suggests possible strategies for improvement of corneal stem cell-based therapy.

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Corneal Endothelial Regeneration Using Mesenchymal Stem Cells Derived from Human Umbilical Cord

Cited by 54 publications

References 43 publications

A new alternative for corneal endothelial regeneration using autologous dental pulp stem cells

A new alternative for corneal endothelial regeneration using autologous dental pulp stem cells

Phenotypic and functional characterization of corneal endothelial cells during in vitro expansion

Corneal Stem Cells as a Source of Regenerative Cell-Based Therapy

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