scholarly journals First-in-human autologous oral mucosal epithelial sheet transplantation to prevent anastomotic re-stenosis in congenital esophageal atresia

2021 ◽  
Author(s):  
Akihiro Fujino ◽  
Yasushi Fuchimoto ◽  
Yoshiyuki Baba ◽  
Nobutaka Isogawa ◽  
Takanori Iwata ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Esophageal Atresia ◽  
Anastomotic Stricture ◽  
Cell Sheet ◽  
Severe Case ◽  
Epithelial Tissue ◽  
Cell Sheets ◽  
Congenital Esophageal Atresia ◽  
Epithelial Sheet ◽  
Oral Epithelial

AbstractBackgroundCongenital esophageal atresia postoperative anastomotic stricture occurs in 30-50% of cases. Patients with severe dysphagia are treated with endoscopic balloon dilatation (EBD) and/or local injection of steroids, but many patients continue to experience frequent stricture. In this study, we investigated the transplantation of autologous oral mucosa-derived cell sheets (epithelial cell sheets) as a prophylactic treatment for congenital esophageal atresia postoperative anastomotic stricture.MethodsEpithelial cell sheets were fabricated from a patient’s oral epithelial tissue, and their safety was confirmed by quality control tests. The epithelial cell sheets were transported under controlled conditions from the fabrication facility to the transplantation facility and successfully transplanted onto the lacerations caused by EBD using a newly developed transplantation device for pediatric patients. The safety of the transplantation was confirmed by follow-up examinations over 48 weeks.ResultsThe number of EBDs required after transplantation and the number of days between EDBs were recorded. Before transplantation, EBDs were performed approximately every two weeks, whereas after transplantation, the interval was extended to a maximum of four weeks. The patient was also aware of a reduction in dysphagia.ConclusionsThis study suggests that cell sheet transplantation might be effective in preventing anastomotic stricture after surgery for congenital esophageal atresia. We chose this very severe case for the first clinical study in humans. Future studies are needed to identify cases in which cell sheet transplantation is most effective and to determine the appropriate timeframes for transplantation.

scholarly journals Keratin intermediate filaments: intermediaries of epithelial cell migration

2019 ◽  
Vol 63 (5) ◽  
pp. 521-533 ◽  
Author(s):  
Sungjun Yoon ◽  
Rudolf E. Leube
Keyword(s):  
Cell Migration ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Biomechanical Properties ◽  
Biochemical Properties ◽  
Cell Junctions ◽  
Epithelial Tissue ◽  
Post Translational Modification ◽  
Cell Sheets ◽  
Epithelial Cell Migration

Abstract Migration of epithelial cells is fundamental to multiple developmental processes, epithelial tissue morphogenesis and maintenance, wound healing and metastasis. While migrating epithelial cells utilize the basic acto-myosin based machinery as do other non-epithelial cells, they are distinguished by their copious keratin intermediate filament (KF) cytoskeleton, which comprises differentially expressed members of two large multigene families and presents highly complex patterns of post-translational modification. We will discuss how the unique mechanophysical and biochemical properties conferred by the different keratin isotypes and their modifications serve as finely tunable modulators of epithelial cell migration. We will furthermore argue that KFs together with their associated desmosomal cell–cell junctions and hemidesmosomal cell–extracellular matrix (ECM) adhesions serve as important counterbalances to the contractile acto-myosin apparatus either allowing and optimizing directed cell migration or preventing it. The differential keratin expression in leaders and followers of collectively migrating epithelial cell sheets provides a compelling example of isotype-specific keratin functions. Taken together, we conclude that the expression levels and specific combination of keratins impinge on cell migration by conferring biomechanical properties on any given epithelial cell affecting cytoplasmic viscoelasticity and adhesion to neighboring cells and the ECM.

scholarly journals Allogeneic Cultivated Limbal Epithelial Sheet Transplantation in Reconstruction of Conjunctival Sac After Chemical and Thermal Burns

2021 ◽  
Vol 8 ◽  
Author(s):  
Ting Wang ◽  
Fengmei Shan ◽  
Qingjun Zhou ◽  
Weiyun Shi ◽  
Lixin Xie
Keyword(s):  
Epithelial Cell ◽  
Cell Sheet ◽  
Corneal Transplantation ◽  
Case Series ◽  
Human Amniotic Membrane ◽  
Safe Alternative ◽  
Thermal Burns ◽  
Palpebral Conjunctiva ◽  
Epithelial Sheet ◽  
Epithelial Sheets

The study aims to evaluate the effect of allogeneic cultivated limbal epithelial cell sheet transplantation (CLET) in reconstructing conjunctival sac for severe symblepharon after chemical and thermal burns. A retrospective, non-comparative case series. Thirty-six eyes (36 patients) underwent CLET for severe symblepharon and conjunctival sac stenosis or atresia. Symblepharon was separated, and pseudopterygium was preserved to replace the palpebral conjunctiva. Allogeneic cultivated limbal epithelial cell sheet using human amniotic membrane as a carrier was transplanted into the recipient's eye to reconstruct the conjunctival sac. The effect of conjunctival sac reconstruction, eye and eyelid movement, ocular surface restitution, and symblepharon recurrence were analyzed after surgery. Symblepharon was completely relieved in 30 of the 36 eyes (83.3%) by a single surgical procedure, with fornix reconstruction, as well as free movement of eye globe and eyelids. Strip-like symblepharon remained in 6 eyes (16.7%) and was completely relieved after the second CLET. Twenty patients without visual function received prostheses 3 months after surgery and the other sixteen patients underwent different corneal transplantation for visual acuity improvement. During the follow-up period, no one had symblepharon recurrence. The transplantation of cultivated allogeneic limbal epithelial sheets offers an effective and safe alternative in the treatment of symblepharon and reconstruction of conjunctival sac in eyes with severe ocular burns, which lays the foundation for subsequent treatments.

scholarly journals ROCK inhibitor combined with Ca2+ controls the myosin II activation and optimizes human nasal epithelial cell sheets

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yoshiyuki Kasai ◽  
Tsunetaro Morino ◽  
Eri Mori ◽  
Kazuhisa Yamamoto ◽  
Hiromi Kojima
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Sheet ◽  
Cell Junctions ◽  
Rock Inhibitor ◽  
Cell Sheets ◽  
Cultured Epithelial Cells ◽  
Rock Inhibition ◽  
Mlc Phosphorylation ◽  
Cell Cell

Abstract The proliferation and differentiation of cultured epithelial cells may be modified by Rho-associated kinase (ROCK) inhibition and extracellular Ca2+ concentration. However, it was not known whether a combination would influence the behavior of cultured epithelial cells through changes in the phosphorylation of non-muscle myosin light chain II (MLC). Here we show that the combination of ROCK inhibition with Ca2+ elevation regulated the phosphorylation of MLC and improved both cell expansion and cell–cell adhesion during the culture of human nasal mucosal epithelial cell sheets. During explant culture, Ca2+ enhanced the adhesion of nasal mucosal tissue, while ROCK inhibition downregulated MLC phosphorylation and promoted cell proliferation. During cell sheet culture, an elevation of extracellular Ca2+ promoted MLC phosphorylation and formation of cell–cell junctions, allowing the harvesting of cell sheets without collapse. Moreover, an in vitro grafting assay revealed that ROCK inhibition increased the expansion of cell sheets three-fold (an effect maintained when Ca2+ was also elevated), implying better wound healing potential. We suggest that combining ROCK inhibition with elevation of Ca2+ could facilitate the fabrication of many types of cell graft.

scholarly journals Partial Decellularized Scaffold Combined with Autologous Nasal Epithelial Cell Sheet for Tracheal Tissue Engineering

2021 ◽  
Vol 22 (19) ◽  
pp. 10322
Author(s):  
Luong Huu Dang ◽  
Shih-Han Hung ◽  
Yuan Tseng ◽  
Ly Xuan Quang ◽  
Nhi Thao Ngoc Le ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Sheet ◽  
Rabbit Model ◽  
Immunohistochemical Analysis ◽  
Airway Epithelial ◽  
Potential Solution ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
Nasal Epithelial Cell

Decellularization has emerged as a potential solution for tracheal replacement. As a fully decellularized graft failed to achieve its purposes, the de-epithelialization partial decellularization protocol appeared to be a promising approach for fabricating scaffolds with preserved mechanical properties and few immune rejection responses after transplantation. Nevertheless, a lack of appropriate concurrent epithelialization treatment can lead to luminal stenosis of the transplant and impede its eventual success. To improve re-epithelialization, autologous nasal epithelial cell sheets generated by our cell sheet engineering platform were utilized in this study under an in vivo rabbit model. The newly created cell sheets have an intact and transplantable appearance, with their specific characteristics of airway epithelial origin being highly expressed upon histological and immunohistochemical analysis. Subsequently, those cell sheets were incorporated with a partially decellularized tracheal graft for autograft transplantation under tracheal partial resection models. The preliminary results two months post operation demonstrated that the transplanted patches appeared to be wholly integrated into the host trachea with adequate healing of the luminal surface, which was confirmed via endoscopic and histologic evaluations. The satisfactory result of this hybrid scaffold protocol could serve as a potential solution for tracheal reconstructions in the future.

scholarly journals Tricellular Tight Junctions in the Inner Ear

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
Shin-ichiro Kitajiri ◽  
Tatsuya Katsuno
Keyword(s):  
Epithelial Cell ◽  
Membrane Proteins ◽  
Inner Ear ◽  
Tight Junctions ◽  
Barrier Function ◽  
Cell Sheet ◽  
Endocochlear Potential ◽  
Cell Sheets ◽  
Current State ◽  
Associated Proteins

Tight junctions (TJs) are structures that seal the space between the epithelial cell sheets. In the inner ear, the barrier function of TJs is indispensable for the separation of the endolymphatic and perilymphatic spaces, which is essential for the generation and maintenance of the endocochlear potential (EP). TJs are formed by the intercellular binding of membrane proteins, known as claudins, and mutations in these proteins cause deafness in humans and mice. Within the epithelial cell sheet, however, a bound structure is present at the site where the corners of three cells meet (tricellular tight junctions (tTJs)), and the maintenance of the barrier function at this location cannot be explained by the claudins alone. Tricellulin and the angulin family of proteins (angulin-1/LSR, angulin-2/ILDR1, and angulin-3/ILDR2) have been identified as tTJ-associated proteins. Tricellulin and ILDR1 are localized at the tTJ and alterations in these proteins have been reported to be involved in deafness. In this review, we will present the current state of knowledge for tTJs.

scholarly journals Biobanked human foreskin epithelial cell sheets reduce inflammation and promote wound healing in a nude mouse model

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Dongliang Zhang ◽  
Jialiang Shao ◽  
Jingming Zhuang ◽  
Shukui Zhou ◽  
Shuo Yin ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Wound Healing ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Sheet ◽  
Skin Defect ◽  
Cell Sheets ◽  
Vascular Regeneration ◽  
Middle Ear Cholesteatoma

Abstract Background Human epithelial cell sheets (ECSs) are used to clinically treat epithelial conditions such as burns, corneal blindness, middle ear cholesteatoma and vitiligo. As a widely used material in clinic, there is little information on the biobanking of ECSs and its repair effect after storage. Results Two methods for biobanking foreskin ECSs were compared in a short term (7 days): 4-degree storage and programmed cryopreservation. Cell sheet integrity, viability, apoptosis, immunogenicity, mechanical properties and function were evaluated. In vivo, ECSs were directly transplanted to skin defect models and histological examination was performed at 1 week postoperatively. We successfully extracted human foreskin-derived primary epithelial cells and fabricated them into ECSs. Compared with 4-degree storage, programmed cryopreservation preserved the ECS structural integrity, enhanced the mechanical properties, decreased HLA-I expression, and increased cell viability and survival. An increased proportion of melanocytes with proliferative capacity remained in the cryopreserved sheets, and the undifferentiated epithelial cells were comparable to those of the fresh sheets. In vivo, cryopreserved ECSs could reduce inflammatory cell infiltration and promote connective tissue remodeling, epithelial cell proliferation and vascular regeneration. Conclusions Programmed cryopreservation of ECSs was superior and more feasible than 4-degree storage and the cryopreserved ECSs achieved satisfying skin wound healing in vivo. We anticipate that the off-the-shelf ECSs could be quickly used, such as, to repair human epithelial defect in future. Graphical abstract

scholarly journals Vinculin is critical for the robustness of the epithelial cell sheet paracellular barrier for ions

2019 ◽  
Vol 2 (4) ◽  
pp. e201900414 ◽  
Author(s):  
Satoshi Konishi ◽  
Tomoki Yano ◽  
Hiroo Tanaka ◽  
Tomoaki Mizuno ◽  
Hatsuho Kanoh ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Atpase Activity ◽  
Barrier Function ◽  
Molecular Mechanisms ◽  
Structural Integrity ◽  
Myosin Ii ◽  
Cell Sheet ◽  
Adherens Junction ◽  
Cell Sheets ◽  
Confluent Cell

The paracellular barrier function of tight junctions (TJs) in epithelial cell sheets is robustly maintained against mechanical fluctuations, by molecular mechanisms that are poorly understood. Vinculin is an adaptor of a mechanosensory complex at the adherens junction. Here, we generated vinculin KO Eph4 epithelial cells and analyzed their confluent cell-sheet properties. We found that vinculin is dispensable for the basic TJ structural integrity and the paracellular barrier function for larger solutes. However, vinculin is indispensable for the paracellular barrier function for ions. In addition, TJs stochastically showed dynamically distorted patterns in vinculin KO cell sheets. These KO phenotypes were rescued by transfecting full-length vinculin and by relaxing the actomyosin tension with blebbistatin, a myosin II ATPase activity inhibitor. Our findings indicate that vinculin resists mechanical fluctuations to maintain the TJ paracellular barrier function for ions in epithelial cell sheets.

A Method for Electron Microscopic Study of Tissue Culture Cell Monolayers Grown in Tubes

1967 ◽  
Vol 25 ◽  
pp. 132-133
Author(s):  
R. Stephens ◽  
G. Schidlovsky ◽  
S. Kuzmic ◽  
P. Gaudreau
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Light Microscopy ◽  
Cell Sheet ◽  
Culture Cell ◽  
Tissue Culture Cell ◽  
Electron Microscopic ◽  
Cell Sheets ◽  
Morphological Alterations ◽  
Culture Cells

The usual method of scraping or trypsinization to detach tissue culture cell sheets from their glass substrate for further pelletization and processing for electron microscopy introduces objectionable morphological alterations. It is also impossible under these conditions to study a particular area or individual cell which have been preselected by light microscopy in the living state.Several schemes which obviate centrifugation and allow the embedding of nondetached tissue culture cells have been proposed. However, they all preserve only a small part of the cell sheet and make use of inverted gelatin capsules which are in this case difficult to handle.We have evolved and used over a period of several years a technique which allows the embedding of a complete cell sheet growing at the inner surface of a tissue culture roller tube. Observation of the same cell by light microscopy in the living and embedded states followed by electron microscopy is performed conveniently.

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