The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Daniel K Podolsky - One of the best experts on this subject based on the ideXlab platform.
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intestinal fibroblasts regulate intestinal Epithelial Cell proliferation via hepatocyte growth factor
American Journal of Physiology-gastrointestinal and Liver Physiology, 1998Co-Authors: Michael Goke, Michiyuki Kanai, Daniel K PodolskyAbstract:Although the presence of subEpithelial intestinal fibroblasts has been well recognized, the effects of fibroblasts on intestinal Epithelial Cell (IEC) growth are incompletely understood. In vitro s...
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hepatocyte growth factor scatter factor modulates intestinal Epithelial Cell proliferation and migration
Biochemical and Biophysical Research Communications, 1994Co-Authors: Axel U Dignass, Kathryn Lynchdevaney, Daniel K PodolskyAbstract:Abstract Various peptide growth factors have been found to regulate Epithelial Cell function within the mucosal epithelium of the gastrointestinal tract. In this study hepatocyte growth factor/scatter factor (HGF/SF) was found to stimulate intestinal Epithelial Cell proliferation: 2.5-fold in the non-transformed rat small intestinal Epithelial Cell line IEC-6 and 1.9-fold in the human colon cancer-derived HT-29 Cell line. In addition, HGF/SF enhanced Epithelial Cell restitution, the initial step involved in gastrointestinal wound healing, in an in vitro model. Migration of IEC-6 in wounded monolayers was enhanced up to 7-fold. Enhancement of restitution by HGF could be completely abrogated by addition of immunoneutralizing anti-TGFβ1, indicating that this process is mediated through a TGFβ-dependent pathway. These findings suggest that HGF exerts functional effects on intestinal Epithelial Cell populations and may play a role in the morphogenesis tract and its remodeling following injury.
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fibroblast growth factors modulate intestinal Epithelial Cell growth and migration
Gastroenterology, 1994Co-Authors: Axel U Dignass, Shoji Tsunekawa, Daniel K PodolskyAbstract:Abstract Background/Aims: Various peptide growth factors have been found to exert functional effects among Epithelial Cell populations. This study assessed the role of certain fibroblast growth factors (FGFs) (acidic FGF, basic FGF, and keratinocyte growth factor) in the regulation of intestinal Epithelial Cell proliferation and restitution. Methods: Recombinant growth factors were added to subconfluent cultures of IEC-6, Caco-2, and HT-29 Cell lines with subsequent assessment of [ 3 H]-thymidine incorporation. The effects on an in vitro model of restitution were assessed by quantitation of Cells migrating into standard wounds established in confluent monolayers of IEC-6 Cells. Transforming growth factor β (TGF-β) content of growth factortreated wounded monolayers was assessed by Northern blot and bioassay. Results: Acidic FGF, basic FGF, and keratinocyte growth factor caused a modest increase in proliferation of IEC-6, Caco-2, and HT-29 Cell lines. Acidic FGF and basic FGF promoted intestinal Epithelial Cell restitution in vitro up to 10-fold, in conjunction with the enhanced expression of TGF-β messenger RNA and protein. Promotion of IEC-6 restitution by acidic and basic FGF could be blocked by addition of immunoneutralizing anti-TGF-β antisera. Conclusions: FGFs that exert effects on fibroblast Cells also exert effects on intestinal Epithelial Cell populations and may help promote Epithelial Cell restitution, the initial step of intestinal wound healing through a TGF-β-dependent pathway.
Kohji Nishida - One of the best experts on this subject based on the ideXlab platform.
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translational research on ocular surface reconstruction using oral mucosal Epithelial Cell sheets
Cornea, 2014Co-Authors: Kohji NishidaAbstract:: Ocular surface reconstruction using autologous oral mucosal Epithelial Cell sheets has drastically changed the treatment of limbal stem-Cell deficiency. The morphological and functional characteristics of oral mucosal Epithelial Cell sheets are similar to those of normal corneal epithelium. Ocular surface reconstruction can prevent potential problems associated with limbal transplantation, including immune rejection and donor tissue shortages. Thus far, ocular reconstruction using Epithelial Cell sheets has been limited to clinical research. Although the effectiveness and safety of this surgical approach have been confirmed to some extent, efforts to make its use more widespread are required. "Translational research" refers to the process of developing a new treatment based on basic research findings with useful practical applications in the field of health care. Medical centers for translational research are required to promote translational research in academic institutes. The Pharmaceutical Affairs Law was revised to promote technologies in the field of regenerative medicine in Japan. This article reviews translational research of ocular surface reconstruction using oral mucosal Epithelial Cell sheets.
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validation system of tissue engineered Epithelial Cell sheets for corneal regenerative medicine
Tissue Engineering Part C-methods, 2010Co-Authors: Ryuhei Hayashi, Masayuki Yamato, Hiroshi Takayanagi, Akira Kubota, Yuichi Hori, Teruo Okano, Kohji NishidaAbstract:Recently, regenerative therapy with tissue-engineered Epithelial Cell sheets has been performed for treating ocular surface disease. It would be required to develop the validation method for these Cell sheets to standardize and spread the regenerative therapy. In the present study, we developed a validation system for cultivated Epithelial Cell sheets. Human limbal Epithelial Cells and human oral mucosal Epithelial Cells were cultured with 3T3 feeder layer Cells on temperature-responsive culture inserts for three different culture periods, and subjected to Cell sheet harvest and validation. Epithelial Cells cultured for a short period were not successfully harvested as intact contiguous Cell sheets. On the other hand, total Cell number and viability of Epithelial Cell sheets harvested after prolonged culture period decreased. Further, these Cells also lost Epithelial barrier function. These results showed the potential effectiveness of the proposed validation system that can evaluate fabricated Cell sheet...
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validation system of tissue engineered Epithelial Cell sheets for corneal regenerative medicine
Tissue Engineering Part C-methods, 2010Co-Authors: Ryuhei Hayashi, Masayuki Yamato, Hiroshi Takayanagi, Akira Kubota, Yuichi Hori, Teruo Okano, Yoshinori Oie, Kohji NishidaAbstract:Recently, regenerative therapy with tissue-engineered Epithelial Cell sheets has been performed for treating ocular surface disease. It would be required to develop the validation method for these Cell sheets to standardize and spread the regenerative therapy. In the present study, we developed a validation system for cultivated Epithelial Cell sheets. Human limbal Epithelial Cells and human oral mucosal Epithelial Cells were cultured with 3T3 feeder layer Cells on temperature-responsive culture inserts for three different culture periods, and subjected to Cell sheet harvest and validation. Epithelial Cells cultured for a short period were not successfully harvested as intact contiguous Cell sheets. On the other hand, total Cell number and viability of Epithelial Cell sheets harvested after prolonged culture period decreased. Further, these Cells also lost Epithelial barrier function. These results showed the potential effectiveness of the proposed validation system that can evaluate fabricated Cell sheets before transplantation.
Teruo Okano - One of the best experts on this subject based on the ideXlab platform.
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bladder augmentation using tissue engineered autologous oral mucosal Epithelial Cell sheets grafted on demucosalized gastric flaps
Transplantation, 2011Co-Authors: Eiko Watanabe, Masayuki Yamato, Yoshiyuki Shiroyanagi, Kazunari Tanabe, Teruo OkanoAbstract:Background At present, autologous intestinal segments are often used for bladder reconstruction. However, the gastrointestinal mucosa often causes various complications. Methods Oral mucosal tissues were obtained from the buccal cavity of beagle dogs. Primary oral mucosal Epithelial Cells were cultured on temperature-responsive culture dishes with a mitomycin C-treated 3T3 feeder layer for 2 weeks. Cultured Epithelial Cells were harvested as contiguous sheets by reducing the temperature to 20°C. The study consisted of three groups. In group 1, oral mucosal Epithelial Cell sheets were autografted on demucosalized gastric flaps. Next, the gastric flaps with the oral mucosal Epithelial Cell sheets were used for bladder reconstruction. Bladder reconstruction was once immediately and then 5 days after Epithelial Cell sheet grafting in groups 2 and 3, respectively. Three weeks after bladder reconstruction, the gastric flaps with the oral mucosal Epithelial Cell sheets were examined by immunohistology. Results Flaps grafted with oral mucosal Epithelial Cell sheets showed Epithelial regeneration in groups 1 and 3. Regenerated epithelia were stratified and similar to native oral mucosa. However, the regenerated epithelium was absent from the reconstructive segment, and urothelial ingrowth was observed in group 2. Macroscopically, all reconstructive segments showed contracture. Conclusions We successfully performed a bladder reconstruction using oral mucosal Epithelial Cell sheet-grafted flaps that exhibited Epithelial regeneration. Further study should consider shrinkage prevention.
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validation system of tissue engineered Epithelial Cell sheets for corneal regenerative medicine
Tissue Engineering Part C-methods, 2010Co-Authors: Ryuhei Hayashi, Masayuki Yamato, Hiroshi Takayanagi, Akira Kubota, Yuichi Hori, Teruo Okano, Kohji NishidaAbstract:Recently, regenerative therapy with tissue-engineered Epithelial Cell sheets has been performed for treating ocular surface disease. It would be required to develop the validation method for these Cell sheets to standardize and spread the regenerative therapy. In the present study, we developed a validation system for cultivated Epithelial Cell sheets. Human limbal Epithelial Cells and human oral mucosal Epithelial Cells were cultured with 3T3 feeder layer Cells on temperature-responsive culture inserts for three different culture periods, and subjected to Cell sheet harvest and validation. Epithelial Cells cultured for a short period were not successfully harvested as intact contiguous Cell sheets. On the other hand, total Cell number and viability of Epithelial Cell sheets harvested after prolonged culture period decreased. Further, these Cells also lost Epithelial barrier function. These results showed the potential effectiveness of the proposed validation system that can evaluate fabricated Cell sheet...
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validation system of tissue engineered Epithelial Cell sheets for corneal regenerative medicine
Tissue Engineering Part C-methods, 2010Co-Authors: Ryuhei Hayashi, Masayuki Yamato, Hiroshi Takayanagi, Akira Kubota, Yuichi Hori, Teruo Okano, Yoshinori Oie, Kohji NishidaAbstract:Recently, regenerative therapy with tissue-engineered Epithelial Cell sheets has been performed for treating ocular surface disease. It would be required to develop the validation method for these Cell sheets to standardize and spread the regenerative therapy. In the present study, we developed a validation system for cultivated Epithelial Cell sheets. Human limbal Epithelial Cells and human oral mucosal Epithelial Cells were cultured with 3T3 feeder layer Cells on temperature-responsive culture inserts for three different culture periods, and subjected to Cell sheet harvest and validation. Epithelial Cells cultured for a short period were not successfully harvested as intact contiguous Cell sheets. On the other hand, total Cell number and viability of Epithelial Cell sheets harvested after prolonged culture period decreased. Further, these Cells also lost Epithelial barrier function. These results showed the potential effectiveness of the proposed validation system that can evaluate fabricated Cell sheets before transplantation.
Yanming Zhang - One of the best experts on this subject based on the ideXlab platform.
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Characteristic and Functional Analysis of a Newly Established Porcine Small Intestinal Epithelial Cell Line
2016Co-Authors: Jing Wang, Zhi Lin, Yanming ZhangAbstract:The mucosal surface of intestine is continuously exposed to both potential pathogens and beneficial commensal microorganisms. Recent findings suggest that intestinal Epithelial Cells, which once considered as a simple physical barrier, are a crucial Cell lineage necessary for maintaining intestinal immune homeostasis. Therefore, establishing a stable and reliable intestinal Epithelial Cell line for future research on the mucosal immune system is necessary. In the present study, we established a porcine intestinal Epithelial Cell line (ZYM-SIEC02) by introducing the human telomerase reverse transcriptase (hTERT) gene into small intestinal Epithelial Cells derived from a neonatal, unsuckled piglet. Morphological analysis revealed a homogeneous cobblestone-like morphology of the Epithelial Cell sheets. Ultrastructural indicated the presence of microvilli, tight junctions, and a glandular configuration typical of the small intestine. Furthermore, ZYM-SIEC02 Cells expressed Epithelial Cell-specific markers including cytokeratin 18, pan-cytokeratin, sucrase-isomaltase, E-cadherin and ZO-1. Immortalized ZYM-SIEC02 Cells remained diploid and were not transformed. In addition, we also examined the host Cell response to Salmonella and LPS and verified the enhanced expression of mRNAs encoding IL-8 and TNF-a by infection with Salmonella enterica serovars Typhimurium (S. Typhimurium). Results showed that IL-8 protein expression were upregulated following Salmonella invasion. TLR4, TLR6 and IL-6 mRNA expression were upregulated following stimulation with LPS, ZYM-SIEC02 Cells were hyporeponsive to LPS with respect to IL-8 mRNA expression and secretion. TNFa mRNA levels wer
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characteristic and functional analysis of a newly established porcine small intestinal Epithelial Cell line
PLOS ONE, 2014Co-Authors: Jingjing Wang, Zhiyi Lin, Yanming ZhangAbstract:The mucosal surface of intestine is continuously exposed to both potential pathogens and beneficial commensal microorganisms. Recent findings suggest that intestinal Epithelial Cells, which once considered as a simple physical barrier, are a crucial Cell lineage necessary for maintaining intestinal immune homeostasis. Therefore, establishing a stable and reliable intestinal Epithelial Cell line for future research on the mucosal immune system is necessary. In the present study, we established a porcine intestinal Epithelial Cell line (ZYM-SIEC02) by introducing the human telomerase reverse transcriptase (hTERT) gene into small intestinal Epithelial Cells derived from a neonatal, unsuckled piglet. Morphological analysis revealed a homogeneous cobblestone-like morphology of the Epithelial Cell sheets. Ultrastructural indicated the presence of microvilli, tight junctions, and a glandular configuration typical of the small intestine. Furthermore, ZYM-SIEC02 Cells expressed Epithelial Cell-specific markers including cytokeratin 18, pan-cytokeratin, sucrase-isomaltase, E-cadherin and ZO-1. Immortalized ZYM-SIEC02 Cells remained diploid and were not transformed. In addition, we also examined the host Cell response to Salmonella and LPS and verified the enhanced expression of mRNAs encoding IL-8 and TNF-α by infection with Salmonella enterica serovars Typhimurium (S. Typhimurium). Results showed that IL-8 protein expression were upregulated following Salmonella invasion. TLR4, TLR6 and IL-6 mRNA expression were upregulated following stimulation with LPS, ZYM-SIEC02 Cells were hyporeponsive to LPS with respect to IL-8 mRNA expression and secretion. TNFα mRNA levels were significantly decreased after LPS stimulation and TNF-α secretion were not detected challenged with S. Typhimurium neither nor LPS. Taken together, these findings demonstrate that ZYM-SIEC02 Cells retained the morphological and functional characteristics typical of primary swine intestinal Epithelial Cells and thus provide a relevant in vitro model system for future studies on porcine small intestinal pathogen-host Cell interactions.
Masayuki Yamato - One of the best experts on this subject based on the ideXlab platform.
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Explant culture of oral mucosal Epithelial Cells for fabricating transplantable Epithelial Cell sheet
Elsevier, 2019Co-Authors: Tsunetaro Morino, Ryo Takagi, Kazuhisa Yamamoto, Hiromi Kojima, Masayuki YamatoAbstract:Introduction: Carrier-free autologous mucosal Epithelial Cell sheets have been clinically utilized as a Cell therapy for various Epithelial disorders. Fabrication of a transplantable oral mucosal Epithelial Cell sheet without mouse feeder layers requires a higher seeding density than that of a sheet with mouse feeder layer culture; therefore, a large amount of donor mucosal tissue is needed. However, Cell grafts co-cultured with mouse feeder layers are classified by the US Food and Drug Administration (FDA) as xenogeneic products. The goal of this study was to evaluate the utility of oral mucosal Epithelial Cells expanded by primary explant culture for the fabrication of an adequate number of transplantable Epithelial Cell sheets without mouse feeder layers. Methods: Small fragments derived from minced oral mucosal tissue were placed into culture dishes for primary explant culture in keratinocyte culture medium. After primary explant culture, the outgrown Cells were treated with trypsin-EDTA and were seeded on a temperature-responsive Cell culture insert. After subculture, the cultured Cells were harvested as a confluent Cell sheet from the culture vessel by temperature reduction. Results: Carrier-free human oral mucosal Epithelial Cell sheets were fabricated in all human cases, and autologous transplantation of the harvested Cell sheets showed rapid Epithelial regeneration to cover Epithelial defects in a rabbit model. The explant culture method, involving the use of small fragments for primary culture, was sufficient for preparing a large number of mucosal Epithelial Cells without mouse feeder layers. Moreover, oral mucosal Epithelial Cells derived from the primary explant culture after cryopreservation allowed for the fabrication of Cell sheets. Conclusions: This method for fabricating transplantable oral mucosal Epithelial Cell sheets is an attractive technique for regenerative medicine. It offers a patient-friendly manufacturing method in which a small amount of biopsy material from the patient represents a sufficient Epithelial Cell source, and a manufacturing plan for preparing Cell grafts can be easily tailored. Keywords: Cell therapy, Regenerative medicine, A small amount of biopsy, A sufficient Epithelial Cell source, Autologous transplantation, Without mouse feeder layer
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bladder augmentation using tissue engineered autologous oral mucosal Epithelial Cell sheets grafted on demucosalized gastric flaps
Transplantation, 2011Co-Authors: Eiko Watanabe, Masayuki Yamato, Yoshiyuki Shiroyanagi, Kazunari Tanabe, Teruo OkanoAbstract:Background At present, autologous intestinal segments are often used for bladder reconstruction. However, the gastrointestinal mucosa often causes various complications. Methods Oral mucosal tissues were obtained from the buccal cavity of beagle dogs. Primary oral mucosal Epithelial Cells were cultured on temperature-responsive culture dishes with a mitomycin C-treated 3T3 feeder layer for 2 weeks. Cultured Epithelial Cells were harvested as contiguous sheets by reducing the temperature to 20°C. The study consisted of three groups. In group 1, oral mucosal Epithelial Cell sheets were autografted on demucosalized gastric flaps. Next, the gastric flaps with the oral mucosal Epithelial Cell sheets were used for bladder reconstruction. Bladder reconstruction was once immediately and then 5 days after Epithelial Cell sheet grafting in groups 2 and 3, respectively. Three weeks after bladder reconstruction, the gastric flaps with the oral mucosal Epithelial Cell sheets were examined by immunohistology. Results Flaps grafted with oral mucosal Epithelial Cell sheets showed Epithelial regeneration in groups 1 and 3. Regenerated epithelia were stratified and similar to native oral mucosa. However, the regenerated epithelium was absent from the reconstructive segment, and urothelial ingrowth was observed in group 2. Macroscopically, all reconstructive segments showed contracture. Conclusions We successfully performed a bladder reconstruction using oral mucosal Epithelial Cell sheet-grafted flaps that exhibited Epithelial regeneration. Further study should consider shrinkage prevention.
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validation system of tissue engineered Epithelial Cell sheets for corneal regenerative medicine
Tissue Engineering Part C-methods, 2010Co-Authors: Ryuhei Hayashi, Masayuki Yamato, Hiroshi Takayanagi, Akira Kubota, Yuichi Hori, Teruo Okano, Kohji NishidaAbstract:Recently, regenerative therapy with tissue-engineered Epithelial Cell sheets has been performed for treating ocular surface disease. It would be required to develop the validation method for these Cell sheets to standardize and spread the regenerative therapy. In the present study, we developed a validation system for cultivated Epithelial Cell sheets. Human limbal Epithelial Cells and human oral mucosal Epithelial Cells were cultured with 3T3 feeder layer Cells on temperature-responsive culture inserts for three different culture periods, and subjected to Cell sheet harvest and validation. Epithelial Cells cultured for a short period were not successfully harvested as intact contiguous Cell sheets. On the other hand, total Cell number and viability of Epithelial Cell sheets harvested after prolonged culture period decreased. Further, these Cells also lost Epithelial barrier function. These results showed the potential effectiveness of the proposed validation system that can evaluate fabricated Cell sheet...
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validation system of tissue engineered Epithelial Cell sheets for corneal regenerative medicine
Tissue Engineering Part C-methods, 2010Co-Authors: Ryuhei Hayashi, Masayuki Yamato, Hiroshi Takayanagi, Akira Kubota, Yuichi Hori, Teruo Okano, Yoshinori Oie, Kohji NishidaAbstract:Recently, regenerative therapy with tissue-engineered Epithelial Cell sheets has been performed for treating ocular surface disease. It would be required to develop the validation method for these Cell sheets to standardize and spread the regenerative therapy. In the present study, we developed a validation system for cultivated Epithelial Cell sheets. Human limbal Epithelial Cells and human oral mucosal Epithelial Cells were cultured with 3T3 feeder layer Cells on temperature-responsive culture inserts for three different culture periods, and subjected to Cell sheet harvest and validation. Epithelial Cells cultured for a short period were not successfully harvested as intact contiguous Cell sheets. On the other hand, total Cell number and viability of Epithelial Cell sheets harvested after prolonged culture period decreased. Further, these Cells also lost Epithelial barrier function. These results showed the potential effectiveness of the proposed validation system that can evaluate fabricated Cell sheets before transplantation.
