The Experts below are selected from a list of 315 Experts worldwide ranked by ideXlab platform

Robert A Stockley - One of the best experts on this subject based on the ideXlab platform.

  • Neutrophil elastase reduces secretion of secretory leukoproteinase inhibitor (SLPI) by lung epithelial Cells: role of charge of the proteinase-inhibitor complex
    Respiratory Research, 2008
    Co-Authors: Anita L Sullivan, Pieter S Hiemstra, Timothy Dafforn, Robert A Stockley
    Abstract:

    Background Secretory leukoproteinase inhibitor (SLPI) is an important inhibitor of neutrophil elastase (NE), a proteinase implicated in the pathogenesis of lung diseases such as COPD. SLPI also has antimicrobial and anti-inflammatory properties, but the concentration of SLPI in lung secretions in COPD varies inversely with infection and the concentration of NE. A fall in SLPI concentration is also seen in Culture supernatants of respiratory Cells exposed to NE, for unknown reasons. We investigated the hypothesis that SLPI complexed with NE associates with Cell membranes in vitro . Methods Respiratory epithelial Cells were Cultured in the presence of SLPI, varying doses of proteinases over time, and in different experimental conditions. The likely predicted charge of the complex between SLPI and proteinases was assessed by theoretical molecular modelling. Results We observed a rapid, linear decrease in SLPI concentration in Culture supernatants with increasing concentration of NE and cathepsin G, but not with other serine proteinases. The effect of NE was inhibited fully by a synthetic NE inhibitor only when added at the same time as NE. Direct contact between NE and SLPI was required for a fall in SLPI concentration. Passive binding to Cell Culture Plate materials was able to remove a substantial amount of SLPI both with and without NE. Theoretical molecular modelling of the structure of SLPI in complex with various proteinases showed a greater positive charge for the complex with NE and cathepsin G than for other proteinases, such as trypsin and mast Cell tryptase, that also bind SLPI but without reducing its concentration. Conclusion These data suggest that NE-mediated decrease in SLPI is a passive, charge-dependent phenomenon in vitro , which may correlate with changes observed in vivo .

  • neutrophil elastase reduces secretion of secretory leukoproteinase inhibitor slpi by lung epithelial Cells role of charge of the proteinase inhibitor complex
    Respiratory Research, 2008
    Co-Authors: Anita L Sullivan, Pieter S Hiemstra, Timothy Dafforn, Robert A Stockley
    Abstract:

    Background: Secretory leukoproteinase inhibitor (SLPI) is an important inhibitor of neutrophil elastase (NE), a proteinase implicated in the pathogenesis of lung diseases such as COPD. SLPI also has antimicrobial and anti-inflammatory properties, but the concentration of SLPI in lung secretions in COPD varies inversely with infection and the concentration of NE. A fall in SLPI concentration is also seen in Culture supernatants of respiratory Cells exposed to NE, for unknown reasons. We investigated the hypothesis that SLPI complexed with NE associates with Cell membranes in vitro. Methods: Respiratory epithelial Cells were Cultured in the presence of SLPI, varying doses of proteinases over time, and in different experimental conditions. The likely predicted charge of the complex between SLPI and proteinases was assessed by theoretical molecular modelling. Results: We observed a rapid, linear decrease in SLPI concentration in Culture supernatants with increasing concentration of NE and cathepsin G, but not with other serine proteinases. The effect of NE was inhibited fully by a synthetic NE inhibitor only when added at the same time as NE. Direct contact between NE and SLPI was required for a fall in SLPI concentration. Passive binding to Cell Culture Plate materials was able to remove a substantial amount of SLPI both with and without NE. Theoretical molecular modelling of the structure of SLPI in complex with various proteinases showed a greater positive charge for the complex with NE and cathepsin G than for other proteinases, such as trypsin and mast Cell tryptase, that also bind SLPI but without reducing its concentration. Conclusion: These data suggest that NE-mediated decrease in SLPI is a passive, charge-dependent phenomenon in vitro, which may correlate with changes observed in vivo.

Teruo Okano - One of the best experts on this subject based on the ideXlab platform.

  • Phenotypic traits of mesenchymal stem Cell sheets fabricated by temperature-responsive Cell Culture Plate: structural characteristics of MSC sheets
    Stem Cell Research & Therapy, 2019
    Co-Authors: Mitsuyoshi Nakao, Kenichi Nagase, David W. Grainger, Hideko Kanazawa, Teruo Okano
    Abstract:

    Background In most stem Cell therapy strategies reported to date, stem Cells are introduced to damaged tissue sites to repair and regenerate the original tissue structure and function. MSC therapeutic efficacies are inconsistent, largely attributed to transplanted MSC difficulties both in engrafting at tissue sites and in retaining their therapeutic functions from suspension formulations. MSC functional components, including Cell adhesion and Cell–Cell junction proteins, and ECM that contribute to essential Cellular therapeutic effects, are damaged or removed by proteolytic enzymes used in stem Cell harvesting strategies from Culture. To overcome these limitations, methods to harvest and transplant Cells without disrupting critical stem Cell functions are required. Cell sheet technology, exploiting temperature-responsive Cell Culture surfaces, permits Cell harvest without Cell protein damage. This study is focused on phenotypic traits of MSC sheets structurally and functionally to understand therapeutic benefits of Cell sheets. Methods/results This study verified cleaved Cellular proteins (vinculin, fibronectin, laminin, integrin β-1, and connexin 43) and increased apoptotic Cell death produced under standard trypsin harvesting treatment in a time-dependent manner. However, MSC sheets produced without trypsin using only temperature-controlled sheet harvest from Culture plastic exhibited intact Cellular structures. Also, MSCs harvested using enzymatic treatment (i.e., chemical disruption) showed higher pYAP expression compared to MSC sheets. Conclusion Retention of Cellular structures such as ECM, Cell–Cell junctions, and Cell–ECM junctions is correlated with human umbilical cord mesenchymal stem Cell (hUC-MSC) survival after detachment from Cell Culture surfaces. Retaining these proteins intact in MSC Cultures using Cell sheet technology is proposed to enhance stem Cell survival and their function in stem Cell-based therapy.

  • Phenotypic traits of mesenchymal stem Cell sheets fabricated by temperature-responsive Cell Culture Plate: structural characteristics of MSC sheets.
    Stem cell research & therapy, 2019
    Co-Authors: Mitsuyoshi Nakao, Kenichi Nagase, David W. Grainger, Hideko Kanazawa, Kyung Sook Kim, Teruo Okano
    Abstract:

    In most stem Cell therapy strategies reported to date, stem Cells are introduced to damaged tissue sites to repair and regenerate the original tissue structure and function. MSC therapeutic efficacies are inconsistent, largely attributed to transplanted MSC difficulties both in engrafting at tissue sites and in retaining their therapeutic functions from suspension formulations. MSC functional components, including Cell adhesion and Cell–Cell junction proteins, and ECM that contribute to essential Cellular therapeutic effects, are damaged or removed by proteolytic enzymes used in stem Cell harvesting strategies from Culture. To overcome these limitations, methods to harvest and transplant Cells without disrupting critical stem Cell functions are required. Cell sheet technology, exploiting temperature-responsive Cell Culture surfaces, permits Cell harvest without Cell protein damage. This study is focused on phenotypic traits of MSC sheets structurally and functionally to understand therapeutic benefits of Cell sheets. This study verified cleaved Cellular proteins (vinculin, fibronectin, laminin, integrin β-1, and connexin 43) and increased apoptotic Cell death produced under standard trypsin harvesting treatment in a time-dependent manner. However, MSC sheets produced without trypsin using only temperature-controlled sheet harvest from Culture plastic exhibited intact Cellular structures. Also, MSCs harvested using enzymatic treatment (i.e., chemical disruption) showed higher pYAP expression compared to MSC sheets. Retention of Cellular structures such as ECM, Cell–Cell junctions, and Cell–ECM junctions is correlated with human umbilical cord mesenchymal stem Cell (hUC-MSC) survival after detachment from Cell Culture surfaces. Retaining these proteins intact in MSC Cultures using Cell sheet technology is proposed to enhance stem Cell survival and their function in stem Cell-based therapy.

Anita L Sullivan - One of the best experts on this subject based on the ideXlab platform.

  • Neutrophil elastase reduces secretion of secretory leukoproteinase inhibitor (SLPI) by lung epithelial Cells: role of charge of the proteinase-inhibitor complex
    Respiratory Research, 2008
    Co-Authors: Anita L Sullivan, Pieter S Hiemstra, Timothy Dafforn, Robert A Stockley
    Abstract:

    Background Secretory leukoproteinase inhibitor (SLPI) is an important inhibitor of neutrophil elastase (NE), a proteinase implicated in the pathogenesis of lung diseases such as COPD. SLPI also has antimicrobial and anti-inflammatory properties, but the concentration of SLPI in lung secretions in COPD varies inversely with infection and the concentration of NE. A fall in SLPI concentration is also seen in Culture supernatants of respiratory Cells exposed to NE, for unknown reasons. We investigated the hypothesis that SLPI complexed with NE associates with Cell membranes in vitro . Methods Respiratory epithelial Cells were Cultured in the presence of SLPI, varying doses of proteinases over time, and in different experimental conditions. The likely predicted charge of the complex between SLPI and proteinases was assessed by theoretical molecular modelling. Results We observed a rapid, linear decrease in SLPI concentration in Culture supernatants with increasing concentration of NE and cathepsin G, but not with other serine proteinases. The effect of NE was inhibited fully by a synthetic NE inhibitor only when added at the same time as NE. Direct contact between NE and SLPI was required for a fall in SLPI concentration. Passive binding to Cell Culture Plate materials was able to remove a substantial amount of SLPI both with and without NE. Theoretical molecular modelling of the structure of SLPI in complex with various proteinases showed a greater positive charge for the complex with NE and cathepsin G than for other proteinases, such as trypsin and mast Cell tryptase, that also bind SLPI but without reducing its concentration. Conclusion These data suggest that NE-mediated decrease in SLPI is a passive, charge-dependent phenomenon in vitro , which may correlate with changes observed in vivo .

  • neutrophil elastase reduces secretion of secretory leukoproteinase inhibitor slpi by lung epithelial Cells role of charge of the proteinase inhibitor complex
    Respiratory Research, 2008
    Co-Authors: Anita L Sullivan, Pieter S Hiemstra, Timothy Dafforn, Robert A Stockley
    Abstract:

    Background: Secretory leukoproteinase inhibitor (SLPI) is an important inhibitor of neutrophil elastase (NE), a proteinase implicated in the pathogenesis of lung diseases such as COPD. SLPI also has antimicrobial and anti-inflammatory properties, but the concentration of SLPI in lung secretions in COPD varies inversely with infection and the concentration of NE. A fall in SLPI concentration is also seen in Culture supernatants of respiratory Cells exposed to NE, for unknown reasons. We investigated the hypothesis that SLPI complexed with NE associates with Cell membranes in vitro. Methods: Respiratory epithelial Cells were Cultured in the presence of SLPI, varying doses of proteinases over time, and in different experimental conditions. The likely predicted charge of the complex between SLPI and proteinases was assessed by theoretical molecular modelling. Results: We observed a rapid, linear decrease in SLPI concentration in Culture supernatants with increasing concentration of NE and cathepsin G, but not with other serine proteinases. The effect of NE was inhibited fully by a synthetic NE inhibitor only when added at the same time as NE. Direct contact between NE and SLPI was required for a fall in SLPI concentration. Passive binding to Cell Culture Plate materials was able to remove a substantial amount of SLPI both with and without NE. Theoretical molecular modelling of the structure of SLPI in complex with various proteinases showed a greater positive charge for the complex with NE and cathepsin G than for other proteinases, such as trypsin and mast Cell tryptase, that also bind SLPI but without reducing its concentration. Conclusion: These data suggest that NE-mediated decrease in SLPI is a passive, charge-dependent phenomenon in vitro, which may correlate with changes observed in vivo.

Mitsuyoshi Nakao - One of the best experts on this subject based on the ideXlab platform.

  • Phenotypic traits of mesenchymal stem Cell sheets fabricated by temperature-responsive Cell Culture Plate: structural characteristics of MSC sheets
    Stem Cell Research & Therapy, 2019
    Co-Authors: Mitsuyoshi Nakao, Kenichi Nagase, David W. Grainger, Hideko Kanazawa, Teruo Okano
    Abstract:

    Background In most stem Cell therapy strategies reported to date, stem Cells are introduced to damaged tissue sites to repair and regenerate the original tissue structure and function. MSC therapeutic efficacies are inconsistent, largely attributed to transplanted MSC difficulties both in engrafting at tissue sites and in retaining their therapeutic functions from suspension formulations. MSC functional components, including Cell adhesion and Cell–Cell junction proteins, and ECM that contribute to essential Cellular therapeutic effects, are damaged or removed by proteolytic enzymes used in stem Cell harvesting strategies from Culture. To overcome these limitations, methods to harvest and transplant Cells without disrupting critical stem Cell functions are required. Cell sheet technology, exploiting temperature-responsive Cell Culture surfaces, permits Cell harvest without Cell protein damage. This study is focused on phenotypic traits of MSC sheets structurally and functionally to understand therapeutic benefits of Cell sheets. Methods/results This study verified cleaved Cellular proteins (vinculin, fibronectin, laminin, integrin β-1, and connexin 43) and increased apoptotic Cell death produced under standard trypsin harvesting treatment in a time-dependent manner. However, MSC sheets produced without trypsin using only temperature-controlled sheet harvest from Culture plastic exhibited intact Cellular structures. Also, MSCs harvested using enzymatic treatment (i.e., chemical disruption) showed higher pYAP expression compared to MSC sheets. Conclusion Retention of Cellular structures such as ECM, Cell–Cell junctions, and Cell–ECM junctions is correlated with human umbilical cord mesenchymal stem Cell (hUC-MSC) survival after detachment from Cell Culture surfaces. Retaining these proteins intact in MSC Cultures using Cell sheet technology is proposed to enhance stem Cell survival and their function in stem Cell-based therapy.

  • Phenotypic traits of mesenchymal stem Cell sheets fabricated by temperature-responsive Cell Culture Plate: structural characteristics of MSC sheets.
    Stem cell research & therapy, 2019
    Co-Authors: Mitsuyoshi Nakao, Kenichi Nagase, David W. Grainger, Hideko Kanazawa, Kyung Sook Kim, Teruo Okano
    Abstract:

    In most stem Cell therapy strategies reported to date, stem Cells are introduced to damaged tissue sites to repair and regenerate the original tissue structure and function. MSC therapeutic efficacies are inconsistent, largely attributed to transplanted MSC difficulties both in engrafting at tissue sites and in retaining their therapeutic functions from suspension formulations. MSC functional components, including Cell adhesion and Cell–Cell junction proteins, and ECM that contribute to essential Cellular therapeutic effects, are damaged or removed by proteolytic enzymes used in stem Cell harvesting strategies from Culture. To overcome these limitations, methods to harvest and transplant Cells without disrupting critical stem Cell functions are required. Cell sheet technology, exploiting temperature-responsive Cell Culture surfaces, permits Cell harvest without Cell protein damage. This study is focused on phenotypic traits of MSC sheets structurally and functionally to understand therapeutic benefits of Cell sheets. This study verified cleaved Cellular proteins (vinculin, fibronectin, laminin, integrin β-1, and connexin 43) and increased apoptotic Cell death produced under standard trypsin harvesting treatment in a time-dependent manner. However, MSC sheets produced without trypsin using only temperature-controlled sheet harvest from Culture plastic exhibited intact Cellular structures. Also, MSCs harvested using enzymatic treatment (i.e., chemical disruption) showed higher pYAP expression compared to MSC sheets. Retention of Cellular structures such as ECM, Cell–Cell junctions, and Cell–ECM junctions is correlated with human umbilical cord mesenchymal stem Cell (hUC-MSC) survival after detachment from Cell Culture surfaces. Retaining these proteins intact in MSC Cultures using Cell sheet technology is proposed to enhance stem Cell survival and their function in stem Cell-based therapy.

Susann M Brady-kalnay - One of the best experts on this subject based on the ideXlab platform.

  • Physically-cross-linked poly(vinyl alcohol) Cell Culture Plate coatings facilitate preservation of Cell-Cell interactions, spheroid formation, and stemness.
    Journal of biomedical materials research. Part B Applied biomaterials, 2021
    Co-Authors: Kathleen Molyneaux, Maria D Wnek, Sonya E L Craig, Jason Vincent, Imani Rucker, Gary E Wnek, Susann M Brady-kalnay
    Abstract:

    We employed aqueous solutions of highly-hydrolyzed (>99+%) poly(vinyl alcohol), PVA, to coat plastic dishes as a method to efficiently induce three-dimensional (3D) culturing of Cells. The coatings were prepared by simple evaporation of 3 wt/vol% solutions of PVA in water and require no additional processing steps after air drying under sterile conditions. The coating allows spheroids to form in solution. Spheroid formation is usually preferable to two-dimensional (2D) culturing as it creates a more realistic ex vivo model of some human tissues and tumors. Using PVA-coated Cell Culture Plates, we demonstrated that we can grow reproducibly sized spheroids using several human glioma Cell lines, including LN229, U87 MG, and Gli36, and the embryonic kidney Cell line, 293T. Spheroids formed on PVA-coated Plates grow as well as on other commercially-available, low-attachment Plates, and have exCellent optical imaging properties. As spheroids, LN229 Cells express markers of cancer stem Cells. Finally, we confirmed that spheroids generated on PVA-coated Plates are sensitive to molecular perturbations, as increased expression of the Cell adhesion molecule PTPμ significantly increased the size of spheroids. The PVA hydrogel layer is an effective tool for creating a more realistic ex vivo Culture system than traditional 2D Culture and can be used to generate Cell spheroids for potential application in drug screening and personalized medicine for diseases such as cancer.