Cultivated Human Conjunctival Epithelial Transplantation for Total Limbal Stem Cell Deficiency

Ang, Leonard P. K.; Tanioka, Hidetoshi; Kawasaki, Satoshi; Ang, Leslie P. S.; Yamasaki, Kenta; Phuc, Tien; Thein, Zaw Moe; Koizumi, Noriko; Nakamura, Takahiro; Yokoi, Norihiko; Komuro, Aoi; Inatomi, Tsutomu; Nakatsukasa, Mina; Kinoshita, Shigeru

doi:10.1167/iovs.09-3379

Cited by 71 publications

(44 citation statements)

References 28 publications

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“…Recently, the use of tissue-engineered epithelial transplantation has revolutionized the treatment of patients with severe ocular surface disease (Ang et al, 2010;Ricardo et al, 2013). The novel use of cultivated autologous conjunctival epithelial transplantation for reconstructing the corneal surface and treating limbal stem cell deficiency is a promising treatment.…”

Section: Discussionmentioning

confidence: 99%

“…The novel use of cultivated autologous conjunctival epithelial transplantation for reconstructing the corneal surface and treating limbal stem cell deficiency is a promising treatment. It has the advantages of safety, a decreased risk of rejection and infection, and no need for long-term treatment with steroids for immunosuppression (Tanioka et al, 2006;Ang et al, 2010). Mucosal integrity of the lacrimal duct system is the key to success using traditional methods of surgery for the correction of lacrimal pathway dysfunctions (traumatic damage, stenosis, and obstruction) (Takahashi et al, 2013;Baran et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…The conjunctival epithelium (CjE), consisting of palpebral, fornical, and bulbar CjE, maintains the microenvironment on the ocular surface. Ex vivo conjunctival grafts have been cultured on amniotic membrane and transplanted into animals and patients with LSCD (Tanioka et al, 2006;Ang et al, 2010;Ricardo et al, 2013). Moreover, the LDE is continuous with the CjE, and expresses a similar broad spectrum of mucins, to reduce drag and enhance tear outflow (Paulsen et al, 2003;Jäger et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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潜在候选细胞构建组织工程的泪道上皮: 兔的组织学和细胞学研究

Xie

Cui

2015

J. Zhejiang Univ. Sci. B

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Abstract:Objective: Injury and deficiency of the lacrimal duct epithelium (LDE) can lead to a variety of lacrimal diseases. The purpose of this study was to characterize potential candidate cells for constructing a tissue-engineered LDE. Methods: Different areas of the conjunctiva and lacrimal duct tissue were removed from male adult New Zealand white rabbits for histological evaluation. Hematoxylin and eosin staining and immunohistochemical staining of cytokeratin AE1+AE3, cytokeratin 4, Ki-67, and MUC5AC were observed by light microscopy. The surface morphologies of different epithelial tissues and cellular structures were examined using field-emission scanning electron microscopy and transmission electron microscopy. Epithelial cells were isolated from tissues and identified by specific markers. In vitro, proliferative ability and Western blot analyses of the proliferating cell nuclear antigen (PCNA) of different epithelial cells cultured in identical environments were investigated and compared. Results: Histologically, the epithelial specific markers, cytokeratin AE1+AE3 and cytokeratin 4, were expressed in the conjunctiva epithelium and the LDE. Notably, highly proliferative cells stained with Ki-67 were concentrated under the epithelium in a dome structure of the posterior palpebral conjunctiva. Differentiated goblet cells were also found to a lesser extent in this region. Primary palpebral and fornical conjunctival epithelial cells (PFCECs), bulbar conjunctival epithelial cells (BCECs), and lacrimal duct epithelial cells (LDECs) were successfully separated from tissues. In vitro, rabbit PFCECs and LDECs grew faster and expressed more PCNA than BCECs. Conclusions: PFCECs are anatomically similar to LDECs. They also have similar morphological characteristics, immune phenotypes, and proliferation features. PFCECs are therefore potential candidate cells to replace LDECs in tissue engineering to treat lacrimal duct diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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潜在候选细胞构建组织工程的泪道上皮: 兔的组织学和细胞学研究

Xie

Cui

2015

J. Zhejiang Univ. Sci. B

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“…Fetal calf serum-supplemented DMEM was chosen as culture media due to good evidence for growth of conjunctival cell lines 19,20 and the same medium is a standard medium in human keratinocytes 8 in combination with human IgG-fractions. Samples were incubated for 12 hours in 5% CO 2 at 378C under the respective conditions.…”

Section: Human Conjunctiva Acquisition and Processingmentioning

confidence: 99%

Pemphigus Autoantibodies Induce Blistering in Human Conjunctiva

Vielmuth

Rötzer

Hartlieb

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. The autoimmune blistering skin disease pemphigus vulgaris (PV) is caused by autoantibodies against desmosomal adhesion molecules. Patients may suffer conjunctival involvement, yet the underlying mechanisms are largely unclear. We characterized human and murine conjunctiva with respect to the PV autoantigens, and evaluated the effects and mechanisms of PV autoantibodies applied to human conjunctiva ex vivo. METHODS.We obtained human conjunctiva specimens from surgical explants and established a short-term culture model to study the alterations induced by antibody fractions of PV patients (PV-IgG). Furthermore, we applied a mouse model depleted of the desmosomal cadherin desmoglein 3 (Dsg3), the primary autoantigen in PV. Murine and human conjunctiva also was used to analyze the expression pattern of desmosomal proteins by immunostaining and Western blotting.RESULTS. Human and murine conjunctiva samples expressed the majority of desmosomal molecules with an expression pattern similar to the epidermis. Interestingly, Dsg3 knock out animals frequently suffer eye lesions, histologically evident as microblisters in the eyelid epidermis and conjunctiva. Incubation of human specimens with PV-IgG for 12 hours caused blistering in the suprabasal layers of the conjunctiva as well as reduction of Dsg1 and Dsg3 protein levels. Furthermore, PV-IgG prompted activation of p38MAPK in the conjunctiva, which is a central pathomechanism leading to blistering in the epidermis.CONCLUSIONS. PV-IgG leads to blister formation and p38MAPK activation in the conjunctiva and, thus, resembles the effects found in the epidermis. Our data indicate that the ocular involvement observed in PV patients is mainly based on conjunctival blistering.

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“…Esses autores também evidenciaram a presença de células caliciformes em culturas de células amplificadoras transitórias, sugerindo uma origem comum à das células epiteliais conjuntivais, porém com diferenciação mais tardia. Como o transplante de células conjuntivais teve sucesso em reconstruir algumas doenças da superfície ocular (29) , Ang et al testaram em modelos de coelhos com deficiência límbica a possibilidade de utilizar a cultura de epitélio conjuntival humano como alternativa ao transplante de epitélio corneano (30) . Eles mostraram que o transplante de células epiteliais da conjuntiva cultivadas ex vivo tinham resultados clínicos equivalentes ao transplante de células epiteliais límbicas cultivadas ex vivo.…”

Section: Conjuntiva Expandidas Ex Vivounclassified

Uso de células-tronco cultivadas ex vivo na reconstrução da superfície ocular

Ricardo

Gomes

2010

Arq. Bras. Oftalmol.

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A s superfícies externas da córnea e da conjuntiva são recobertas por epitélios altamente especializados, que são derivados de células precursoras genotipicamente diversas. Ambos são epitélios do tipo escamoso estratificado não-queratinizado, formados por camadas de células que continuamente se renovam e se regeneram. As camadas basais contêm células-tronco (CTs) semiquiescentes capazes de autoregeneração e também de criar outras células que se diferenciam e migram para a superfície externa, quando cessa a divisão celular (1) . Esses epitélios são separados entre si por uma zona de transição conhecida como limbo. Acredita-se que as paliçadas de Vogt localizadas no limbo contenham estruturas chamadas criptas epiteliais límbicas que representam o nicho das CTs ABSTRACTLesions on the ocular surface can destroy the stem cells from the limbus and cause limbal stem cell deficiency. The limbal stem cell deficiency is marked by conjunctivalization, which can be defined as the invasion of conjunctival epithelium over the cornea. This process is accompanied by varying degrees of corneal changes such as neovascularization, inflammation, recurrent erosions, persistent epithelial defects, destruction of basement membrane of epithelium and stromal healing. Often, these changes are associated with poor visual acuity, photophobia and ocular discomfort. The best treatment for this disease is not known and varies in unilateral or bilateral cases. Among the treatments available, transplantation of limbal autograft or allograft is one of the most used. To improve the outcome of allotransplantation, some researchers use the transplantation of corneal epithelium cultured in the laboratory by ex vivo expansion of limbal stem cells, but due to limited availability of autologous tissue from the limbus and the risk of complications associated with immunosuppression in allogeneic tissue transplantation, researches of others options of stem cell cultured ex vivo have been described in experimental and clinical stage. This review describes the new types of stem cells cultured ex vivo, their current results and future potential. RESUMOLesões na superfície ocular podem atingir as células-tronco do limbo e causar deficiência límbica. A deficiência límbica é caracterizada pela conjuntivalização, que pode ser definida como a invasão do epitélio conjuntival sobre a córnea. Este processo é acompanhado por graus variáveis de alterações corneanas, como neovascularização, inflamação, erosões recorrentes, defeitos epiteliais persistentes, destruição da membrana basal do epitélio e cicatrização estromal. Frequentemente, estas alterações estão associadas à diminuição da acuidade visual, fotofobia e desconforto ocular. O melhor tratamento para essa afecção não é conhecido e possibilidades variam em casos uni ou bilaterais. Entre os tratamentos disponíveis, o transplante de limbo autólogo ou alógeno é um dos mais utilizados. Para melhorar os resultados dos transplantes alógenos, alguns pesquisadores utilizam o transplante de epitélio da córnea cultivado...

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Cultivated Human Conjunctival Epithelial Transplantation for Total Limbal Stem Cell Deficiency

Abstract: The study showed that microscopically, HCjE cells have features similar to HCE cells, with clinically equivalent outcomes. The ex vivo cultivation of conjunctiva to form transplantable epithelial sheets for corneal replacement is a promising new treatment modality in patients with LSCD.

Cited by 71 publications

References 28 publications

潜在候选细胞构建组织工程的泪道上皮: 兔的组织学和细胞学研究

潜在候选细胞构建组织工程的泪道上皮: 兔的组织学和细胞学研究

Pemphigus Autoantibodies Induce Blistering in Human Conjunctiva

Uso de células-tronco cultivadas ex vivo na reconstrução da superfície ocular

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