Evaluation of <scp>ABCG</scp>2 and p63 expression in canine cornea and cultivated corneal epithelial cells

Morita, Maresuke; Fujita, Naoki; Takahashi, Ayaka; Nam, Eun Ryel; Yui, Sho; Chung, Cheng Shu; Kawahara, Naoya; Lin, Hsing Yi; Tsuzuki, Keiko; Nakagawa, Takayuki; Nishimura, Ryohei

doi:10.1111/vop.12147

Cited by 32 publications

(29 citation statements)

References 27 publications

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“…Of these markers, p63 has been consistently found in basal and suprabasal cells in the limbus of horses, goats, rabbits, dogs, humans, and lab animals . Conflicting results, however, have been reported for the center of the cornea: no p63 expression has been detected in rabbits, only sporadically in suprabasal cells of goats and, consistent with our results, positive basal and suprabasal cells have been found throughout the corneal epithelium in humans, dogs, and horses . Our results support the interpretation of Schlötzer‐Schrehardt and Kruse, that basal p63 positivity may represent cells in proliferative state such as TACs and only include the corneal epithelial SC population.…”

Section: Discussionsupporting

confidence: 91%

“…In accordance with the human cornea, we found NGF in basal cells of noncrypt, limbal, and peripheral zones, however, additional positive cells were present in the center. Consistent results have been described for ABCG2 expression of basal and suprabasal cells of limbus of humans, dogs, and rabbits . However, conflicting results have been reported regarding the central cornea, since ABCG2 was shown to be absent in dogs and rabbits, but in agreement with our findings positive cells have been found in humans .…”

Section: Discussionsupporting

confidence: 91%

“…Consistent results have been described for ABCG2 expression of basal and suprabasal cells of limbus of humans, dogs, and rabbits . However, conflicting results have been reported regarding the central cornea, since ABCG2 was shown to be absent in dogs and rabbits, but in agreement with our findings positive cells have been found in humans . Therefore, a conclusion corresponding to p63 can be drawn for NGF and ABCG2.…”

Section: Discussionsupporting

confidence: 91%

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Morphological and immunohistochemical characteristics of the equine corneal epithelium

Kammergruber

Rahn

Nell

et al. 2019

Veterinary Ophthalmology

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ObjectiveThe morphology of the corneal epithelium in two age groups of horses is described. Distribution patterns of proliferation‐, differentiation‐, stem cell‐associated markers and cell junction proteins were assessed.MethodsCorneal samples from 12 horses (six foals and six adult horses) were analyzed after H&E staining and immunohistochemistry using the following antibodies: E‐cadherin, β‐catenin, Connexin 43 (Cx43), tight junction protein 1 (TJP1), cytokeratin (CK) 14, CK 19, CK 3, CK 10, vimentin, Ki67, p63, nerve growth factor (NGF), ABCG2, and epithelial growth factor receptor. Semiquantitative analysis of crypt, limbal, peripheral, and central zone was performed. Semithin and ultrathin sections were used for ultrastructural evaluation of the epithelium.ResultsThe height of the epithelium varied between age groups and crypts were consistently present. In the peripheral and central epithelium, three types of basal cells resembling a pseudostratified epithelium were characterized. Potential stem cell markers (CK 14, p63, NGF, and ABCG2) were present in all zones with decreasing frequency toward the center. Cornea‐specific differentiation marker CK 3 was not expressed in the most basal cell layer of the limbal epithelium. E‐cadherin, β‐catenin, and Cx43 revealed a similar apico‐lateral signal pattern throughout the entire epithelium; only TJP1 was additionally seen at the basal surface.ConclusionsThis study presents a systematic semiquantitative evaluation of the equine corneal epithelium, showing the presence of crypts as potential stem cell niche with CK 14, p63, NGF, and ABCG2 as relevant markers for cells with regenerative capacity. The pseudostratified arrangement of the basal layer was a unique finding.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 91%

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Morphological and immunohistochemical characteristics of the equine corneal epithelium

Kammergruber

Rahn

Nell

et al. 2019

Veterinary Ophthalmology

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“…ABCG-2 has a role in the maintenance of retinal stem cells under the regulation of Notch signalling. The expression of ABCG-2 in canine limbal epithelial cells has been considered a useful marker for presence of putative corneal epithelial stem cells and proliferation [34,35]. The ex vivo 3D tissue engineered pterygium outgrowths and the graft-containing cultures expressed all of these stem cell markers (Sox2, Oct4 and ABCG-2).…”

Section: Discussionmentioning

confidence: 99%

Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy

Josifovska

Szabó

Nagymihály

et al. 2017

Contact Lens and Anterior Eye

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A B S T R A C TPurpose: Development of ex vivo model to study pathogenesis, inflammation and treatment modalities for pterygium. Methods: Pterygium obtained from surgery was cultivated (3 months). Gravitational attachment method using viscoelastic facilitated adherence of graft and outgrowing cells. Medium contained serum as the only growth supplement with no use of scaffolds. Surface profiling of the multi-layered cells for hematopoietic-and mesenchymal stem cell markers was performed. Examination of cells by immunohistochemistry using pluripotency, oxidative stress, stemness, migration and proliferation, epithelial and secretory markers was performed. The effect of anti-proliferative agent Mitomycin C upon secretion of pro-inflammatory cytokines IL-6 and IL-8 was assessed. Results: Cells showed high expression of migration-(CXCR4), secretory-(MUC1, MUC4) and oxidative damage-(8-OHdG) markers, and low expression of hypoxia-(HIF-1α) and proliferation-(Ki-67) markers. Moderate and low expression of the pluripotency markers (Vimentin and ΔNp63) was present, respectively, while the putative markers of stemness (Sox2, Oct4, ABCG-2) and epithelial cell markers-(CK19, CK8-18) were weak. The surface marker profile of the outgrowing cells revealed high expression of the hematopoietic marker CD47, mesenchymal markers CD90 and CD73, minor or less positivity for the hematopoietic marker CD34, mesenchymal marker CD105, progenitor marker CD117 and attachment protein markers while low levels of IL-6 and IL-8 secretion ex vivo, were inhibited upon Mitomycin C treatment. Conclusion: Ex vivo tissue engineered pterygium consists of a mixture of cells of different lineage origin, suitable for use as a disease model for studying pathogenesis ex vivo, while opening possibilities for new treatment and prevention modalities.

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“…By searching for markers of epithelial progenitors or stem cells in the endometria (Westfall et al, 2003;Cykowski et al, 2016;Morita et al, 2015), we found that ΔNp63 mRNA level in AS group was dramatically upregulated by 139-folds of the controls ( Figure 1D). Through case to control comparison, 3/10 cases of AS showed overlap with controls on ΔNp63 transcription ( Figure 1D).…”

Section: δNp63 Upregulation Is Associated With Endometrial Quiescencementioning

confidence: 99%

Transplantation of collagen scaffold with autologous bone marrow mononuclear cells promotes functional endometrium reconstruction via downregulating ΔNp63 expression in Asherman’s syndrome

Zhao

Cao

Zhu

et al. 2016

Sci. China Life Sci.

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Asherman's syndrome (AS) is a common disease that presents endometrial regeneration disorder. However, little is known about its molecular features of this aregenerative endometrium in AS and how to reconstruct the functioning endometrium for the patients with AS. Here, we report that ΔNp63 is significantly upregulated in residual epithelial cells of the impaired endometrium in AS; the upregulated-ΔNp63 induces endometrial quiescence and alteration of stemness. Importantly, we demonstrate that engrafting high density of autologous bone marrow mononuclear cells (BMNCs) loaded in collagen scaffold onto the uterine lining of patients with AS downregulates ΔNp63 expression, reverses ΔNp63-induced pathological changes, normalizes the stemness alterations and restores endometrial regeneration. Finally, five patients achieved successful pregnancies and live births. Therefore, we conclude that ΔNp63 is a crucial therapeutic target for AS. This novel treatment significantly improves the outcome for the patients with severe AS.Asherman's syndrome, ΔNp63, quiescence, endometrial regeneration, bone marrow stem cell based therapy Citation:

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Evaluation of ABCG2 and p63 expression in canine cornea and cultivated corneal epithelial cells

Abstract: ABCG2 was localized in canine limbal epithelial cells, and p63 was widely expressed in canine corneal epithelia. ABCG2 and p63 could prove to be useful markers in dogs for putative corneal epithelial stem cells and for corneal epithelial cell proliferation, respectively.

Cited by 32 publications

References 27 publications

Morphological and immunohistochemical characteristics of the equine corneal epithelium

Morphological and immunohistochemical characteristics of the equine corneal epithelium

Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy

Transplantation of collagen scaffold with autologous bone marrow mononuclear cells promotes functional endometrium reconstruction via downregulating ΔNp63 expression in Asherman’s syndrome

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