Long-term Observation After Transplantation of Cultured Human Corneal Endothelial Cells for Corneal Endothelial Dysfunction

Abstract Background At present, corneal transplantation is still the only way to treat serious corneal diseases caused by corneal endothelial dysfunction. However, the shortage of donor cornea tissues and human corneal endothelial cells (HCECs) remains a worldwide challenge. We cultivated HCECs by the use of a conditioned medium from orbital adipose-derived stem cells (OASC-CM) in vitro. Then the HCECs were used to treat animal corneal endothelial dysfunction models via cell transplantation. The initial effect was gratifying. The purpose of this study was to conduct a long-term observation and evaluation after cell transplantation. Methods First, orbital adipose-derived stem cells (OASCs) were isolated to prepare conditioned medium (CM). Then HCECs were cultivated and expanded by the usage of CM (CM-HCECs). Related CEC markers were analyzed by immunofluorescence. Cells proliferation ability was also tested. CM-HCECs were then transplanted into monkey corneal endothelial dysfunction models by cell injection. We carried out a 24-month postoperative preclinical observation and verified the long-term effect by histological examination and transcriptome sequencing. Results CM-HCECs expressed HCEC related markers and maintained polygonal cell morphology after several passages. During 24 months of cell transplantation into the monkey's anterior chamber, the cornea thickness and transparency kept healthy status, and the corneal endothelial cell density remained in the normal range. Gene sequencing showed that the gene expression pattern of CM-HCECs was similar to that of transplanted cells and HCECs. Conclusions The proliferation and repair ability of HCECs were significantly improved due to the effect of OASC-CM. The result of this study confirmed long-term therapeutic efficacy of CM-HCECs in vivo. Our research provided an extensive cell source and a promising prospect for regenerative medicine and cell-based therapy.

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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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Transplantation of Human Corneal Endothelial Cells Cultured on Bio-Engineered Collagen Vitrigel in a Rabbit Model of Corneal Endothelial Dysfunction

Current Eye Research ◽

10.1080/02713683.2017.1351568 ◽

2017 ◽

Vol 42 (11) ◽

pp. 1420-1425 ◽

Cited By ~ 8

Author(s):

Junko Yoshida ◽

Seiichi Yokoo ◽

Ayumi Oshikata-Miyazaki ◽

Shiro Amano ◽

Toshiaki Takezawa ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Dysfunction ◽

Rabbit Model ◽

Corneal Endothelial Cells ◽

Human Corneal Endothelial Cells ◽

Corneal Endothelial Dysfunction ◽

Cells Cultured ◽

Collagen Vitrigel

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Conditioned Medium from Human Umbilical Vein Endothelial Cells Promotes Proliferation, Migration, Invasion and Angiogenesis of Adipose Derived Stem Cells

Current Medical Science ◽

10.1007/s11596-018-1855-8 ◽

2018 ◽

Vol 38 (1) ◽

pp. 124-130 ◽

Cited By ~ 3

Author(s):

Ming-lian Luo ◽

Xiao-ping Liu ◽

Fang Wang ◽

Xiao-xia Liu ◽

Wei-fang Liu ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Conditioned Medium ◽

Umbilical Vein ◽

Adipose Derived Stem Cells ◽

Human Umbilical Vein ◽

Umbilical Vein Endothelial Cells

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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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Paracrine effect of human adipose-derived stem cells on lymphatic endothelial cells

Regenerative Medicine ◽

10.2217/rme-2020-0071 ◽

2020 ◽

Vol 15 (9) ◽

pp. 2085-2098

Author(s):

Cristiana Marcozzi ◽

Annalisa Frattini ◽

Marina Borgese ◽

Federica Rossi ◽

Ludovica Barone ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Conditioned Medium ◽

Lymphatic Vessels ◽

Lymphatic Endothelial Cell ◽

Adipose Derived Stem Cells ◽

Lymphatic Endothelial Cells ◽

Paracrine Effect ◽

Secondary Lymphedema

Aim: The proposal of this study was to evaluate, in vitro, the potential paracrine effect of human adipose-derived stem cells (hASCs) to promote lymphangiogenesis in lymphatic endothelial cells isolated from rat diaphragmatic lymphatic vessels. Materials & methods: ELISA on VEGFA, VEGFC and IL6 in hASC-conditioned medium; LYVE1 immunostaining; and gene expression of PROX1, VEGFR3, VEGFC, VEGFA and IL6 were the methods used. Results: In 2D culture, hASC-conditioned medium was able to promote lymphatic endothelial cell survival, maintenance of endothelial cobblestone morphology and induction to form a vessel-like structure. Conclusion: The authors' results represent in vitro evidence of the paracrine effect of hASCs on lymphatic endothelial cells, suggesting the possible role of hASC-conditioned medium in developing new therapeutic approaches for lymphatic system-related dysfunction such as secondary lymphedema.

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In vitro Three Dimensional Scaffold-free Construct of Human Adipose-derived Stem Cells in Coculture with Endothelial Cells and Fibroblasts

Revista de Chimie ◽

10.37358/rc.17.6.5670 ◽

2017 ◽

Vol 68 (6) ◽

pp. 1341-1344

Author(s):

Grigore Berea ◽

Gheorghe Gh. Balan ◽

Vasile Sandru ◽

Paul Dan Sirbu

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Endothelial Cells ◽

Single Cell ◽

Cell Line ◽

Bone Repair ◽

Umbilical Vein ◽

Human Fibroblasts ◽

Three Dimensional ◽

Adipose Derived Stem Cells

Complex interactions between stem cells, vascular cells and fibroblasts represent the substrate of building microenvironment-embedded 3D structures that can be grafted or added to bone substitute scaffolds in tissue engineering or clinical bone repair. Human Adipose-derived Stem Cells (hASCs), human umbilical vein endothelial cells (HUVECs) and normal dermal human fibroblasts (NDHF) can be mixed together in three dimensional scaffold free constructs and their behaviour will emphasize their potential use as seeding points in bone tissue engineering. Various combinations of the aforementioned cell lines were compared to single cell line culture in terms of size, viability and cell proliferation. At 5 weeks, viability dropped for single cell line spheroids while addition of NDHF to hASC maintained the viability at the same level at 5 weeks Fibroblasts addition to the 3D construct of stem cells and endothelial cells improves viability and reduces proliferation as a marker of cell differentiation toward osteogenic line.

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