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Article ; Online: Donor Corneal Endothelial Cell Maturity and Its Impact on Graft Survival in Glaucoma Patients Undergoing Corneal Transplantation.

Kitazawa, Koji / Toda, Munetoyo / Ueno, Morio / Wakimasu, Koichi / Tomioka, Yasufumi / Uehara, Asako / Sotozono, Chie / Kinoshita, Shigeru

American journal of ophthalmology

2024 Volume 262, Page(s) 1–9

Abstract: Purpose: To examine corneal graft survival via corneal endothelial cell density (ECD) and corneal endothelial cell loss (ECL) at 5 years post-transplantation in the eyes of patients with and without a history of undergoing glaucoma surgery according to ... ...

Abstract	Purpose: To examine corneal graft survival via corneal endothelial cell density (ECD) and corneal endothelial cell loss (ECL) at 5 years post-transplantation in the eyes of patients with and without a history of undergoing glaucoma surgery according to the maturity of the donor corneal endothelial cells. Design: Prospective cohort study. Methods: This prospective cohort study included 17 patients with glaucoma and 51 patients without glaucoma who underwent Descemet's stripping automated endothelial keratoplasty or penetrating keratoplasty at the Baptist Eye Institute, Kyoto, Japan, between October 2014 and October 2016. Human corneal endothelial cells were cultured from residual peripheral donor cornea tissue, and the maturity of the cells was evaluated by cell surface markers (ie, CD166 Results: At 36 months postoperatively, the mean ECD and ECL in the glaucoma-bleb eyes were 1197 ± 352 cells/mm Conclusions: The findings in this prospective cohort study revealed that the use of donor corneal grafts containing mature-differentiated corneal endothelial cells could maintain the survival of the transplanted graft for a long-term period, even in patients with a history of undergoing glaucoma surgery.
MeSH term(s)	Humans ; Graft Survival/physiology ; Prospective Studies ; Male ; Female ; Endothelium, Corneal/pathology ; Aged ; Tissue Donors ; Cell Count ; Middle Aged ; Intraocular Pressure/physiology ; Glaucoma/surgery ; Glaucoma/physiopathology ; Corneal Endothelial Cell Loss/diagnosis ; Keratoplasty, Penetrating ; Descemet Stripping Endothelial Keratoplasty ; Follow-Up Studies ; Flow Cytometry ; Aged, 80 and over ; Visual Acuity/physiology
Language	English
Publishing date	2024-02-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	80030-2
ISSN	1879-1891 ; 0002-9394
ISSN (online)	1879-1891
ISSN	0002-9394
DOI	10.1016/j.ajo.2024.01.033
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Article ; Online: The Biologic Character of Donor Corneal Endothelial Cells Influences Endothelial Cell Density Post Successful Corneal Transplantation.

Kitazawa, Koji / Toda, Munetoyo / Ueno, Morio / Uehara, Asako / Sotozono, Chie / Kinoshita, Shigeru

Ophthalmology science

2022 Volume 3, Issue 2, Page(s) 100239

Abstract: Purpose: Corneal endothelial cell density (ECD) gradually decreases after corneal transplantation by unknown biologic, biophysical, or immunologic mechanism. Our purpose was to assess the association between donor corneal endothelial cell (CEC) maturity ...

Abstract	Purpose: Corneal endothelial cell density (ECD) gradually decreases after corneal transplantation by unknown biologic, biophysical, or immunologic mechanism. Our purpose was to assess the association between donor corneal endothelial cell (CEC) maturity in culture and postoperative endothelial cell loss (ECL) after successful corneal transplantation. Design: Prospective cohort study. Participants: This cohort study was conducted at Baptist Eye Institute, Kyoto, Japan, between October 2014 and October 2016. It included 68 patients with a 36-month follow-up period who had undergone successful Descemet stripping automated endothelial keratoplasty (DSAEK) or penetrating keratoplasty. Methods: Human CECs (HCECs) from remaining peripheral donor corneas were cultured and evaluated for maturity by surface markers (CD166 Main outcome measures: Endothelial cell density and ECL at 36 months postoperative. Results: The 68 included patients (mean [standard deviation] age 68.1 [13.6] years, 47.1% women, 52.9% DSAEK). The high, middle, and low-maturity groups included 17, 32, and 19 eyes, respectively. At 36 months postoperative, the mean (standard deviation) ECD significantly decreased to 911 (388) cells/mm Conclusions: The high content of mature-differentiated HCECs expressed in culture by the donor peripheral cornea was coincident with low ECL, suggesting that a high-maturity CEC content predicts long-term graft survival. Understanding the molecular mechanism for maintaining HCEC maturity could elucidate the mechanism of ECL after corneal transplantation and aid in developing effective interventions. Financial disclosures: Proprietary or commercial disclosure may be found after the references.
Language	English
Publishing date	2022-10-28
Publishing country	Netherlands
Document type	Journal Article
ISSN	2666-9145
ISSN (online)	2666-9145
DOI	10.1016/j.xops.2022.100239
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Article ; Online: Quiescent innate and adaptive immune responses maintain the long-term integrity of corneal endothelium reconstituted through allogeneic cell injection therapy.

Toda, Munetoyo / Ueno, Morio / Yamada, Jun / Hiraga, Asako / Asada, Kazuko / Hamuro, Junji / Sotozono, Chie / Kinoshita, Shigeru

Scientific reports

2022 Volume 12, Issue 1, Page(s) 18072

Abstract: This study aims to clarify the immunogenicity in acquired and innate immune responses of cultured human corneal endothelial cells (hCECs) applied for cell injection therapy, a newly established modality for corneal endothelium failures. Thirty-four ... ...

Abstract	This study aims to clarify the immunogenicity in acquired and innate immune responses of cultured human corneal endothelial cells (hCECs) applied for cell injection therapy, a newly established modality for corneal endothelium failures. Thirty-four patients with corneal endothelial failure received injection of allogeneic hCEC suspension into anterior chamber. No sign of immunological rejection was observed in all 34 patients during the 5-8 years postoperative follow-up period. Cell injection therapy was successful in 2 patients treated for endothelial failure after penetrating keratoplasty and one patient with Descemet membrane stripping automated endothelial keratoplasty failure. ELISPOT assays performed in allo-mixed lymphocyte reaction to the alloantigen identical to that on the injected hCECs, elicited sparse IFN-γ-specific spots in the peripheral blood mononuclear cells of patients who received hCEC injection. The therapy generated simple and smooth graft-host junctions without wound stress. The injection of C57BL/6 CECs into the anterior chamber of BALB/c mice, which rejected C57BL/6 corneas 6 weeks ago, induced no sign of inflammatory reactions after the second challenge of alloantigen. Collectively, injection of the hCEC cell suspension in the aqueous humor induces immune tolerance that contributes to the survival of the reconstituted endothelium.
MeSH term(s)	Mice ; Animals ; Humans ; Endothelium, Corneal ; Allogeneic Cells ; Endothelial Cells ; Leukocytes, Mononuclear ; Mice, Inbred C57BL ; Mice, Inbred BALB C ; Isoantigens ; Immunity ; Corneal Diseases/surgery
Chemical Substances	Isoantigens
Language	English
Publishing date	2022-10-27
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-22522-4
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Article ; Online: In Vivo Fluorescence Visualization of Anterior Chamber Injected Human Corneal Endothelial Cells Labeled With Quantum Dots.

Toda, Munetoyo / Yukawa, Hiroshi / Yamada, Jun / Ueno, Morio / Kinoshita, Shigeru / Baba, Yoshinobu / Hamuro, Junji

Investigative ophthalmology & visual science

2019 Volume 60, Issue 12, Page(s) 4008–4020

Abstract: Purpose: The injection of cultured human corneal endothelial cells (cHCECs) into the anterior chamber (AC) is a newly developed modality for the successful treatment of corneal endothelium dysfunction. Here, we investigated whether or not cHCECs could ... ...

Abstract	Purpose: The injection of cultured human corneal endothelial cells (cHCECs) into the anterior chamber (AC) is a newly developed modality for the successful treatment of corneal endothelium dysfunction. Here, we investigated whether or not cHCECs could be labeled using quantum dots (QDs) composed of semiconductor nanoparticle octa-arginine (R8) to trace injected cHCECs and examined the utility of in vivo fluorescence imaging to analyze the dynamics and accumulation of QD-labeled injected cHCECs in a corneal endothelial dysfunction mouse model. Methods: The cHCECs, either of high quality or with cell-state transition, were labeled by adding a mixture of QDs655 and R8. The labeling efficiency and the unchanging of the cell phenotypes by the labeling was confirmed by flow cytometry. The labeled cHCECs were injected into the AC of either healthy mice or mice with corneal endothelium damaged by cryogenic treatment. The kinetics of the injected cHCECs was traced quantitatively via multiphoton confocal laser microscopy. Results: QD labeling induced no morphologic change in the cHCECs or in the expression of the functional markers of cHCECs (i.e., Na+/K+-ATPase and zonula occludens-1). The injected cHCECs-QDs were quantitatively detected, and the retention of cHCECs-QDs was evident, from 3 to 48 hours post cell injection on the posterior surface in the cryogenically injured corneal endothelium mouse model eyes, yet not in the noninjured healthy control eyes. Conclusions: The findings of this study show that in the field of regenerative medicine, QD labeling of cells presents a convenient and sensitive method of finely monitoring the fate of injected cells in vivo.
MeSH term(s)	Adult ; Animals ; Anterior Chamber/metabolism ; Biomarkers/metabolism ; Cell Count ; Cell Differentiation ; Cells, Cultured ; Endothelium, Corneal/metabolism ; Flow Cytometry ; Humans ; Injections, Intraocular ; Male ; Mice ; Mice, Inbred C57BL ; Microscopy, Fluorescence ; Microscopy, Phase-Contrast ; Quantum Dots ; Regenerative Medicine/methods ; Sodium-Potassium-Exchanging ATPase/metabolism ; Tissue Donors ; Young Adult ; Zonula Occludens-1 Protein/metabolism
Chemical Substances	Biomarkers ; Tjp1 protein, mouse ; Zonula Occludens-1 Protein ; Sodium-Potassium-Exchanging ATPase (EC 7.2.2.13)
Language	English
Publishing date	2019-09-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.19-27788
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Article ; Online: Repressed miR-34a Expression Dictates the Cell Fate to Corneal Endothelium Failure.

Hamuro, Junji / Asada, Kazuko / Ueno, Morio / Yamashita, Tomoko / Mukai, Atsushi / Fujita, Tomoko / Ito, Eiko / Hiramoto, Nao / Toda, Munetoyo / Sotozono, Chie / Kinoshita, Shigeru

Investigative ophthalmology & visual science

2022 Volume 63, Issue 4, Page(s) 22

Abstract: Purpose: To reveal the mechanism triggering the functional disparity between degenerated and non-degenerated corneal endothelium cells in the water efflux from corneal stroma to the anterior chamber.: Methods: The varied levels of the microRNA (miR)- ... ...

Abstract	Purpose: To reveal the mechanism triggering the functional disparity between degenerated and non-degenerated corneal endothelium cells in the water efflux from corneal stroma to the anterior chamber. Methods: The varied levels of the microRNA (miR)-34, miR-378, and miR-146 family in human corneal endothelium and cultured cells thereof were investigated using 3D-Gene Human miRNA Oligo Chips. Concomitantly, CD44, p53, c-Myc, matrix metalloprotease (MMP)-2 expression, and Ras homolog gene family member A (Rho A) activity was correlated to the expression intensities of these microRNAs, partly complemented with their altered expression levels with the transfection of the corresponding mimics and inhibitors. The levels of miRs were further associated with intracellular pH (pHi) and mitochondrial energy homeostasis. Results: P53-inducible miR-34a/b repressed CD44 expression, and CD44 was repressed with the elevated c-Myc. The repressed miR-34a activated the CD44 downstream factors Rho A and MMP-2. MiR-34a mimics downregulated pHi, inducing the skewing of mitochondrial respiration to oxidative phosphorylation. The oxidative stress (H2O2) induced on human corneal endothelial cells, which repressed miR-34a/b expression, may account for the impaired signaling cascade to mitochondrial metabolic homeostasis necessary for an efficient water efflux from the corneal stroma. Conclusions: The upregulated expression of CD44, through repressed miR-34a/b by reactive oxygen species and elevated c-Myc by oxidative stress, may impair mitochondrial metabolic homeostasis, leading to human corneal endothelial failure.
MeSH term(s)	Endothelial Cells/metabolism ; Endothelium, Corneal/metabolism ; Humans ; Hydrogen Peroxide/metabolism ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Water/metabolism
Chemical Substances	MIRN34 microRNA, human ; MicroRNAs ; Water (059QF0KO0R) ; Hydrogen Peroxide (BBX060AN9V)
Language	English
Publishing date	2022-04-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.63.4.22
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Article ; Online: A physical biomarker of the quality of cultured corneal endothelial cells and of the long-term prognosis of corneal restoration in patients.

Yamamoto, Akihisa / Tanaka, Hiroshi / Toda, Munetoyo / Sotozono, Chie / Hamuro, Junji / Kinoshita, Shigeru / Ueno, Morio / Tanaka, Motomu

Nature biomedical engineering

2019 Volume 3, Issue 12, Page(s) 953–960

Abstract: Dysfunction of the corneal endothelium reduces the transparency of the cornea and can cause blindness. Because corneal endothelial cells have an extremely limited proliferative ability in vivo, treatment for corneal endothelial dysfunction involves the ... ...

Abstract	Dysfunction of the corneal endothelium reduces the transparency of the cornea and can cause blindness. Because corneal endothelial cells have an extremely limited proliferative ability in vivo, treatment for corneal endothelial dysfunction involves the transplantation of donor corneal tissue. Corneal endothelium can also be restored via intraocular injection of endothelial cells in suspension after their expansion in vitro. Yet, because quality assessment during the expansion of the cells is a destructive process, a substantial number of the cultured cells are lost. Here, we show that the 'spring constant' of the effective interaction potential between endothelial cells in a confluent monolayer serves as a biomarker of the quality of corneal endothelial cells in vitro and of the long-term prognosis of corneal restoration in patients treated with culture-expanded endothelial cells or with transplanted corneas. The biomarker can be measured from phase contrast imaging in vitro and from specular microscopy in vivo, and may enable a shift from passive monitoring to pre-emptive intervention in patients with severe corneal disorders.
MeSH term(s)	Adolescent ; Adult ; Aged ; Biomarkers ; Cell Culture Techniques/methods ; Cells, Cultured ; Cornea/cytology ; Cornea/diagnostic imaging ; Cornea/surgery ; Corneal Diseases/diagnosis ; Corneal Diseases/pathology ; Corneal Diseases/therapy ; Corneal Transplantation/methods ; Endothelial Cells/cytology ; Endothelial Cells/transplantation ; Endothelium, Corneal/cytology ; Endothelium, Corneal/diagnostic imaging ; Endothelium, Corneal/transplantation ; Humans ; Middle Aged ; Prognosis ; Young Adult
Chemical Substances	Biomarkers
Language	English
Publishing date	2019-07-22
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2157-846X
ISSN (online)	2157-846X
DOI	10.1038/s41551-019-0429-9
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Article ; Online: Superiority of Mature Differentiated Cultured Human Corneal Endothelial Cell Injection Therapy for Corneal Endothelial Failure.

Ueno, Morio / Toda, Munetoyo / Numa, Kohsaku / Tanaka, Hiroshi / Imai, Kojiro / Bush, John / Teramukai, Satoshi / Okumura, Naoki / Koizumi, Noriko / Yamamoto, Akihisa / Tanaka, Motomu / Sotozono, Chie / Hamuro, Junji / Kinoshita, Shigeru

American journal of ophthalmology

2021 Volume 237, Page(s) 267–277

Abstract: Purpose: To investigate the safety and efficacy of cultured human corneal endothelial cell (hCEC) injection therapy with mature differentiated (mature) cell subpopulations (SPs) for corneal endothelial failure (CEF).: Design: Comparative, ... ...

Abstract	Purpose: To investigate the safety and efficacy of cultured human corneal endothelial cell (hCEC) injection therapy with mature differentiated (mature) cell subpopulations (SPs) for corneal endothelial failure (CEF). Design: Comparative, interventional case series. Methods: This study involved 18 eyes with CEF that underwent cultured hCEC injection therapy, categorized into 2 groups: (1) 11 eyes administered a relatively lower proportion (0.1 to 76.3%) of mature cell SPs (group 1 [Gr1]), and (2) 7 eyes administered a relatively higher proportion (>90%) of mature cell SPs (group 2 [Gr2]). From 1 week to 3 years postoperation, corneal endothelial cell (CEC) density (CECD), central corneal thickness (CCT), and best-corrected visual acuity (BCVA) were recorded, and the CEC parameter's "spring constant" was calculated. The proportion of mature SPs was evaluated by fluorescence-activated cell sorting analysis based on cell-surface markers. Results: At 3 years postoperation, corneal restoration with improved BCVA was attained in 10 of the 11 Gr1 eyes and all Gr2 eyes, the median CECD in Gr2 (3083 cells/mm Conclusion: Our findings showed that mature cell SPs for hCEC injection therapy provide rapid recovery of CCT, better CECD, and low CECD attrition over 3 years postsurgery.
MeSH term(s)	Cell Count ; Cell Differentiation ; Cells, Cultured ; Cornea ; Endothelial Cells ; Endothelium, Corneal ; Humans
Language	English
Publishing date	2021-11-14
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80030-2
ISSN	1879-1891 ; 0002-9394
ISSN (online)	1879-1891
ISSN	0002-9394
DOI	10.1016/j.ajo.2021.11.012
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Article ; Online: Metabolic Plasticity in Cell State Homeostasis and Differentiation of Cultured Human Corneal Endothelial Cells.

Hamuro, Junji / Ueno, Morio / Asada, Kazuko / Toda, Munetoyo / Montoya, Monty / Sotozono, Chie / Kinoshita, Shigeru

Investigative ophthalmology & visual science

2016 Volume 57, Issue 10, Page(s) 4452–4463

Abstract: Purpose: To clarify whether cultured human corneal endothelial cells (cHCECs), heterogeneous in their differentiation state, exhibit distinctive energy metabolism with the aim to develop a reliable method to sort cHCECs applicable for regenerative ... ...

Abstract	Purpose: To clarify whether cultured human corneal endothelial cells (cHCECs), heterogeneous in their differentiation state, exhibit distinctive energy metabolism with the aim to develop a reliable method to sort cHCECs applicable for regenerative medicine. Methods: The presence of cHCEC subpopulations (SPs) was verified via surface cluster-of-differentiation (CD) marker expression. Cultured HCEC metabolic extracts or corresponding culture supernatants with distinctive cellular phenotypes in regard to energy-metabolism-related functional markers c-Myc and CD44 were prepared and analyzed via capillary electrophoresis-tandem mass spectrometry. The metabolic requirements of heterogeneous SPs of cHCECs were also investigated. Results: After successfully discriminating SPs, as verified via surface CD markers in terms of their secretory metabolites, we found that the CD44+++ SP with cell-state transition (CST) exhibited disposition for anaerobic glycolysis, whereas the CD44-SP without CST was disposed to mitochondria-dependent oxidative phosphorylation (OXPHOS). These results raised the possibility of establishing effective culture conditions to selectively expand mature cHCECs with a hexagonal cobblestone shape and inclination for mitochondria-dependent OXPHOS. Conclusions: The findings of this study open a pathway for monitoring the disposition of cHCECs via their energy metabolism, thus leading to safe and stable regenerative medicine by use of metabolically defined cHCECs in cell-suspension form.
MeSH term(s)	Adolescent ; Adult ; Aged ; Biomarkers/metabolism ; Cell Differentiation ; Cell Plasticity/physiology ; Cells, Cultured ; Child ; Child, Preschool ; Corneal Diseases/diagnosis ; Corneal Diseases/metabolism ; Electrophoresis, Capillary ; Endothelium, Corneal/metabolism ; Endothelium, Corneal/pathology ; Energy Metabolism/physiology ; Female ; Flow Cytometry ; Homeostasis ; Humans ; Male ; Middle Aged ; Tandem Mass Spectrometry ; Young Adult
Chemical Substances	Biomarkers
Language	English
Publishing date	2016-10-18
Publishing country	United States
Document type	Journal Article
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.16-19807
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Article ; Online: Cultured Human Corneal Endothelial Cell Aneuploidy Dependence on the Presence of Heterogeneous Subpopulations With Distinct Differentiation Phenotypes.

Hamuro, Junji / Ueno, Morio / Toda, Munetoyo / Sotozono, Chie / Montoya, Monty / Kinoshita, Shigeru

Investigative ophthalmology & visual science

2016 Volume 57, Issue 10, Page(s) 4385–4392

Abstract: Purpose: Cultured human corneal endothelial cells (cHCECs) are anticipated to become an alternative to donor corneas for the treatment of corneal endothelial dysfunction. However, cHCECs reportedly tend to exhibit chromosomal abnormality during in vitro ...

Abstract	Purpose: Cultured human corneal endothelial cells (cHCECs) are anticipated to become an alternative to donor corneas for the treatment of corneal endothelial dysfunction. However, cHCECs reportedly tend to exhibit chromosomal abnormality during in vitro cell division, thereby hampering their use in the clinical setting. The purpose of this study was to clarify whether a specified subpopulation (SP) of heterogeneous cHCECs would exhibit aneuploidy, whereas other SPs would not. Methods: The presence of SPs in cHCECs was analyzed on the basis of surface cluster of differentiation (CD) antigen CD166, CD105, CD44, CD26, and CD24 expression levels by flow cytometry. Cytogenetic examination was performed for 23 lots of cHCECs, either as whole-cell preparations (bulk) consisting of mixed SPs or as a semipurified SP by magnetic activated cell sorting (MACS). The HCEC donors ranged from 9 to 69 years of age and the culture passages from primary to fifth passage. Results: Flow cytometry analysis demonstrated the presence of at least three cHCEC SPs. One SP, purified by MACS, with surface expression of CD166+, CD105-, CD44-, CD24-, and CD26- did not show any aneuploidy in 50 cells. However, CD166+, CD44+++, CD24-, and CD26+ cHCEC SPs showed sex chromosome loss in all cells (60 cells), whereas CD166+, CD44+++, CD24+, and CD26- SPs exhibited, albeit partly, trisomy on chromosomes 6, 7, 12, and 20. Conclusions: We found that cHCEC aneuploidy is linked to specified SPs present in cHCECs and that the SP sharing the surface phenotype with mature HCECs in corneal tissues was devoid of the karyotype abnormality.
MeSH term(s)	Adolescent ; Adult ; Aged ; Aneuploidy ; Cell Adhesion Molecules/metabolism ; Cell Differentiation ; Cell Division ; Cells, Cultured ; Child ; Corneal Endothelial Cell Loss/genetics ; Corneal Endothelial Cell Loss/pathology ; Corneal Endothelial Cell Loss/surgery ; Corneal Transplantation/methods ; Endothelium, Corneal/cytology ; Endothelium, Corneal/metabolism ; Endothelium, Corneal/transplantation ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Phenotype ; Tissue Donors ; Young Adult
Chemical Substances	Cell Adhesion Molecules
Language	English
Publishing date	2016-08-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.16-19771
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Article ; Online: Allogeneic Sensitization and Tolerance Induction After Corneal Endothelial Cell Transplantation in Mice.

Yamada, Jun / Ueno, Morio / Toda, Munetoyo / Shinomiya, Katsuhiko / Sotozono, Chie / Kinoshita, Shigeru / Hamuro, Junji

Investigative ophthalmology & visual science

2016 Volume 57, Issue 11, Page(s) 4572–4580

Abstract: Purpose: We evaluated the allogeneic response after corneal endothelial cell transplantation in the anterior chamber (AC) in a new mouse model by examining the acquisition of a delayed-type hypersensitivity (DTH) response, induction of allogeneic AC- ... ...

Abstract	Purpose: We evaluated the allogeneic response after corneal endothelial cell transplantation in the anterior chamber (AC) in a new mouse model by examining the acquisition of a delayed-type hypersensitivity (DTH) response, induction of allogeneic AC-associated immune deviation (ACAID), and acquisition of delayed transplantation tolerance. Method: The corneal eyecups from C57BL/6 mice were prepared. The epithelial layer was detached with EDTA solution and treated with trypsin to release mouse-derived primary corneal endothelial cells (mpCECs). The mpCECs (1 × 104 cells) were transplanted into the AC of the eye or subcutaneously (SC) into the neck of BALB/c mice. In the mouse model of endothelial cell transplantation, the endothelial cells in a 2-mm central area of the cornea were eliminated by cryoinjury. The mpCEC transplant model was evaluated by measuring allogeneic cell survival and corneal thickness. The allospecific DTH response and ACAID induction were evaluated 1 week after transplantation. The long-term transplantation tolerance was evaluated by observing a secondary penetrating keratoplasty (PKP) performed on the same donor C57BL/6 mice. Results: The SC injection of mpCECs induced a DTH response, whereas the AC injection induced ACAID. However, eyes inflamed by cryoinjury showed neither the DTH response nor ACAID following AC injection. The mpCECs survived for at least 1 week after injection. Penetrating keratoplasty allografts at 8 weeks after mpCEC transplantation survived indefinitely (100%). Conclusions: The mpCECs display low allogenicity in the AC and are capable of inducing allogeneic tolerance. Corneal endothelial cell transplantation into the AC may represent a safe technique for allogeneic transplantation.
MeSH term(s)	Animals ; Anterior Chamber/immunology ; Cell Transplantation/methods ; Corneal Edema/pathology ; Corneal Edema/surgery ; Corneal Transplantation/methods ; Disease Models, Animal ; Endothelium, Corneal/cytology ; Endothelium, Corneal/transplantation ; Graft Survival/immunology ; Hypersensitivity, Delayed/immunology ; Hypersensitivity, Delayed/pathology ; Immune Tolerance ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Transplantation, Homologous
Language	English
Publishing date	2016-09-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	391794-0
ISSN	1552-5783 ; 0146-0404
ISSN (online)	1552-5783
ISSN	0146-0404
DOI	10.1167/iovs.15-19020
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