Heterologous Transplantation Versus Enhancement of Human Corneal Endothelium

Insler, Michael S.; Lopez, Jamie G.

doi:10.1097/00003226-199103000-00009

Cited by 42 publications

(27 citation statements)

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“…[30][31][32][33][34][35] Cultured HCECs expanded in an in vitro culture system can be composed of multiple SPs of cHCECs, including those with CST. Cultured cells also tend toward karyotype changes.…”

Section: Discussionmentioning

confidence: 99%

Cell Homogeneity Indispensable for Regenerative Medicine by Cultured Human Corneal Endothelial Cells

Hamuro

Toda

Asada

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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Citation: Hamuro J, Toda M, Asada K, et al. Cell homogeneity indispensable for regenerative medicine by cultured human corneal endothelial cells. Invest Ophthalmol Vis Sci. 2016;57:4749-4761. DOI:10.1167/ iovs.16-19770 PURPOSE. To identify the subpopulation (SP) among heterogeneous cultured human corneal endothelial cells (cHCECs) devoid of cell-state transition applicable for cell-based therapy.METHODS. Subpopulation presence in cHCECs was confirmed via surface CD-marker expression level by flow cytometry. CD markers effective for distinguishing distinct SPs were selected by analyzing those on established cHCECs with a small cell area and high cell density. Contrasting features among three typical cHCEC SPs was confirmed by PCR array for extracellular matrix (ECM). Combined analysis of CD markers was performed to identify the SP (effector cells) applicable for therapy. ZO-1 and Na þ /K þ ATPase, CD200, and HLA expression were compared among heterogeneous SPs. RESULTS. Flow cytometry analysis identified the effector cell expressing CD166 and the presence of other SPs with CD166þþþ (CD26 and CD24, either þ or À) was confirmed. PCR array revealed three distinct ECM expression profiles. Some SPs expressed ZO-1 and Naþ ATPase at comparable levels with effector cells, while only one SP expressed CD200, but not on effector cells. Human leukocyte antigen expression was most reduced in the effector SP. The proportion of effector cells (E-ratio) inversely paralleled donor age and decreased during prolonged culture passages. The presence of Rho-associated protein kinase (ROCK) inhibitor increased the E-ratio in cHCECs. The average area of effector cells was approximately 200~220 lm 2 , and the density of cHCECs exceeded 2500 cells/mm 2 . CONCLUSIONS.A specified cultured effector cell population sharing the surface phenotypes with mature HCECs in corneal tissues may serve as an alternative to donor corneas for the treatment of corneal endothelial dysfunction.

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Section: Discussionmentioning

confidence: 99%

Cell Homogeneity Indispensable for Regenerative Medicine by Cultured Human Corneal Endothelial Cells

Hamuro

Toda

Asada

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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“…11,[41][42][43][44][45] One of the most notable obstacles to the application of cHCECs for cell-based therapy is the lack of an established reliable method for verifying the quality of cHCECs. In this study, we presented a noninvasive method for monitoring cell quality, as opposed to the invasive method that involves following surface CD markers.…”

Section: Discussionmentioning

confidence: 99%

Metabolic Plasticity in Cell State Homeostasis and Differentiation of Cultured Human Corneal Endothelial Cells

Hamuro

Ueno

Asada

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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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-ofdifferentiation (CD) marker expression. Cultured HCEC metabolic extracts or corresponding culture supernatants with distinctive cellular phenotypes in regard to energy-metabolismrelated 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 CD44þþþ SP with cell-state transition (CST) exhibited disposition for anaerobic glycolysis, whereas the CD44 À 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.

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“…The ability to demonstrate in vitro Descemet's reconstitution using a bovine system suggested that endothelial decompensation could be reversed by the replacement of normal endothelial cells [1]. Such as approach would lower the antigenic load compared with whole cornea re- Insler and Lopez [5] were able to demonstrate a 67% success rate in transplants of donor corneas denuded and reconstituted in vitro with cultured human fetal CEC and transplanted into nonhuman primates. The high level of success is a strong indication that such a reconstitution protocol is workable.…”

Section: Discussionmentioning

confidence: 99%

“…Several groups have initiated studies to replace defective corneal endothelial cells (CEC) by either: (1) seeding CEC directly onto the donor cornea, which is then transplanted [1,3,5,7], or (2) using a carrier, either an artificial membrane or a natural carrier such as Descemet's membrane [8,9], to allow cells to migrate onto the cornea. The preliminary results indicate that these methodologies may be suitable alternatives to penetrating keratoplasty in some cases.…”

Section: Introductionmentioning

confidence: 99%

Repopulation of denuded murine Descemet's membrane with life-extended murine corneal endothelial cells as a model for corneal cell transplantation

Joo

Green

Pepose³

et al. 2000

Graefe's Archive for Clinical and Experimental Ophthalmology

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This experimental protocol establishes that "life-extended" MCEC can bind to Descemet's membrane ex vivo and form a distinct monolayer. The repopulated Descemet's membrane allowed us to directly test the hypothesis that cultured life-extended corneal endothelial cells are functional when reintroduced into an in vivo milieu and provides evidence that specific corneal endothelial cell transplantation may be a viable alternative to pentrating keratoplasty.

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Heterologous Transplantation Versus Enhancement of Human Corneal Endothelium

Cited by 42 publications

References 0 publications

Cell Homogeneity Indispensable for Regenerative Medicine by Cultured Human Corneal Endothelial Cells

Cell Homogeneity Indispensable for Regenerative Medicine by Cultured Human Corneal Endothelial Cells

Metabolic Plasticity in Cell State Homeostasis and Differentiation of Cultured Human Corneal Endothelial Cells

Repopulation of denuded murine Descemet's membrane with life-extended murine corneal endothelial cells as a model for corneal cell transplantation

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