The Experts below are selected from a list of 315 Experts worldwide ranked by ideXlab platform
Yukio Imanishi - One of the best experts on this subject based on the ideXlab platform.
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Photoimmobilization of insulin onto polystyrene dishes for protein-Free Cell culture.
Biotechnology progress, 1996Co-Authors: Yoshihiro Ito, Guoping Chen, Yukio ImanishiAbstract:Photoreactive insulin was synthesized by coupling with azidobenzoic acid. The insulin derivative was immobilized onto the wells of polystyrene culture plates by photoir-radiation. The photoimmobilized insulin enhanced the growth of anchorage-dependent Cells such as Chinese hamster ovary CHO-K1 and mouse fibroblast STO Cells by more than native or azidophenyl-derivatized insulin. A small amount of photoimmobilized insulin (1-10% of the amount of the native or derivatized insulin) enhanced the growth of CHO-K1 and STO Cells. In addition, the maximal mitogenic effect of the immobilized insulin was greater than that of native or derivatized insulin. Photoimmobilization could be a universal means of immobilizing growth factors onto the surface of materials devoid of chemical functional groups scaffolding growth factors and providing a new protein-Free Cell culture system or tissue engineering materials.
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protein Free Cell culture on an artificial substrate with covalently immobilized insulin
Proceedings of the National Academy of Sciences of the United States of America, 1996Co-Authors: Yoshihiro Ito, Ji Zheng, Yukio Imanishi, Kazuyoshi Yonezawa, Masato KasugaAbstract:Abstract Insulin was immobilized on a surface-hydrolyzed poly(methyl methacrylate) film. Chinese hamster ovary Cells overexpressing human insulin receptors were cultured on the film in the absence of serum or soluble proteins. Small amounts of immobilized insulin (1-10% of the required amount of Free insulin) were sufficient to stimulate Cell proliferation. In addition, the maximal mitogenic effect of immobilized insulin was greater than that of Free insulin. Immobilized insulin activated the insulin receptor and downstream signaling proteins, and this activation persisted for longer periods than that obtained with Free insulin, probably explaining the greater mitogenic effect of the immobilized insulin. Finally the immobilized-insulin film was usable repeatedly without marked loss of activity.
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serum Free Cell culture on insulin immobilized porous collagen beads
Biotechnology and Bioengineering, 1995Co-Authors: Yoshihiro Ito, Ji Zheng, Yukio ImanishiAbstract:Insulin or albumin was immobilized on collagen beads using water-soluble carbodiimide. Adhesion of STO mouse fibroblast Cells onto the beads decreased with increasing the amount of immobilized proteins. Growth of the Cells was remarkably accelerated on the insulinimmobilized collagen beads, which can be used for serum-Free Cell culture. The growth acceleration became larger with increasing the amount of immobilized insulin, while it became smaller with increasing the amount of immobilized albumin. In addition, the immobilized insulin more strongly accelerated the Cell growth than Free insulin plus collagen beads. (c) 1995 John Wiley & Sons, Inc.
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Serum‐Free Cell culture on insulin‐immobilized porous collagen beads
Biotechnology and bioengineering, 1995Co-Authors: Yoshihiro Ito, Ji Zheng, Yukio ImanishiAbstract:Insulin or albumin was immobilized on collagen beads using water-soluble carbodiimide. Adhesion of STO mouse fibroblast Cells onto the beads decreased with increasing the amount of immobilized proteins. Growth of the Cells was remarkably accelerated on the insulinimmobilized collagen beads, which can be used for serum-Free Cell culture. The growth acceleration became larger with increasing the amount of immobilized insulin, while it became smaller with increasing the amount of immobilized albumin. In addition, the immobilized insulin more strongly accelerated the Cell growth than Free insulin plus collagen beads. (c) 1995 John Wiley & Sons, Inc.
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Cell growth on immobilized Cell growth factor 8 protein Free Cell culture on insulin immobilized microcarriers
Biotechnology and Bioengineering, 1992Co-Authors: Yoshihiro Ito, Tsuyoshi Uno, Shu Qin Liu, Yukio ImanishiAbstract:In order to develop a new protein-Free Cell culture system, microcarriers immobilized with insulin were synthesized. For the synthesis, glass and polyacrylamide beads were treated for the introduction of amino groups on the surface, and insulin was immobilized on the surface by using several method. Anchorage-dependent Cells. mouse fibroblast Cells STO and fibroic sarcoma Cells HSDM(1)C(1), and the anchorage-independent Cells, mouse hybridoma Cells SJK132-20 and RDP 45/20 were cultivated on the microcarriers immobilized with insulin. The insulin-immobilized microcarriers did not have any effect on the proliferation of the anchorage independent Cells but promoted the growth of anchorage-dependent Cells remarkably. The activity of immobilized insulin was larger than that of Free or adsorbed insulin. The repeated use of the insulin-immobilized microcarrier was possible, and the promotion activity in the the repeated use was greater than that in the use.
Yoshihiro Ito - One of the best experts on this subject based on the ideXlab platform.
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Photoimmobilization of insulin onto polystyrene dishes for protein-Free Cell culture.
Biotechnology progress, 1996Co-Authors: Yoshihiro Ito, Guoping Chen, Yukio ImanishiAbstract:Photoreactive insulin was synthesized by coupling with azidobenzoic acid. The insulin derivative was immobilized onto the wells of polystyrene culture plates by photoir-radiation. The photoimmobilized insulin enhanced the growth of anchorage-dependent Cells such as Chinese hamster ovary CHO-K1 and mouse fibroblast STO Cells by more than native or azidophenyl-derivatized insulin. A small amount of photoimmobilized insulin (1-10% of the amount of the native or derivatized insulin) enhanced the growth of CHO-K1 and STO Cells. In addition, the maximal mitogenic effect of the immobilized insulin was greater than that of native or derivatized insulin. Photoimmobilization could be a universal means of immobilizing growth factors onto the surface of materials devoid of chemical functional groups scaffolding growth factors and providing a new protein-Free Cell culture system or tissue engineering materials.
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protein Free Cell culture on an artificial substrate with covalently immobilized insulin
Proceedings of the National Academy of Sciences of the United States of America, 1996Co-Authors: Yoshihiro Ito, Ji Zheng, Yukio Imanishi, Kazuyoshi Yonezawa, Masato KasugaAbstract:Abstract Insulin was immobilized on a surface-hydrolyzed poly(methyl methacrylate) film. Chinese hamster ovary Cells overexpressing human insulin receptors were cultured on the film in the absence of serum or soluble proteins. Small amounts of immobilized insulin (1-10% of the required amount of Free insulin) were sufficient to stimulate Cell proliferation. In addition, the maximal mitogenic effect of immobilized insulin was greater than that of Free insulin. Immobilized insulin activated the insulin receptor and downstream signaling proteins, and this activation persisted for longer periods than that obtained with Free insulin, probably explaining the greater mitogenic effect of the immobilized insulin. Finally the immobilized-insulin film was usable repeatedly without marked loss of activity.
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serum Free Cell culture on insulin immobilized porous collagen beads
Biotechnology and Bioengineering, 1995Co-Authors: Yoshihiro Ito, Ji Zheng, Yukio ImanishiAbstract:Insulin or albumin was immobilized on collagen beads using water-soluble carbodiimide. Adhesion of STO mouse fibroblast Cells onto the beads decreased with increasing the amount of immobilized proteins. Growth of the Cells was remarkably accelerated on the insulinimmobilized collagen beads, which can be used for serum-Free Cell culture. The growth acceleration became larger with increasing the amount of immobilized insulin, while it became smaller with increasing the amount of immobilized albumin. In addition, the immobilized insulin more strongly accelerated the Cell growth than Free insulin plus collagen beads. (c) 1995 John Wiley & Sons, Inc.
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Serum‐Free Cell culture on insulin‐immobilized porous collagen beads
Biotechnology and bioengineering, 1995Co-Authors: Yoshihiro Ito, Ji Zheng, Yukio ImanishiAbstract:Insulin or albumin was immobilized on collagen beads using water-soluble carbodiimide. Adhesion of STO mouse fibroblast Cells onto the beads decreased with increasing the amount of immobilized proteins. Growth of the Cells was remarkably accelerated on the insulinimmobilized collagen beads, which can be used for serum-Free Cell culture. The growth acceleration became larger with increasing the amount of immobilized insulin, while it became smaller with increasing the amount of immobilized albumin. In addition, the immobilized insulin more strongly accelerated the Cell growth than Free insulin plus collagen beads. (c) 1995 John Wiley & Sons, Inc.
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Cell growth on immobilized Cell growth factor 8 protein Free Cell culture on insulin immobilized microcarriers
Biotechnology and Bioengineering, 1992Co-Authors: Yoshihiro Ito, Tsuyoshi Uno, Shu Qin Liu, Yukio ImanishiAbstract:In order to develop a new protein-Free Cell culture system, microcarriers immobilized with insulin were synthesized. For the synthesis, glass and polyacrylamide beads were treated for the introduction of amino groups on the surface, and insulin was immobilized on the surface by using several method. Anchorage-dependent Cells. mouse fibroblast Cells STO and fibroic sarcoma Cells HSDM(1)C(1), and the anchorage-independent Cells, mouse hybridoma Cells SJK132-20 and RDP 45/20 were cultivated on the microcarriers immobilized with insulin. The insulin-immobilized microcarriers did not have any effect on the proliferation of the anchorage independent Cells but promoted the growth of anchorage-dependent Cells remarkably. The activity of immobilized insulin was larger than that of Free or adsorbed insulin. The repeated use of the insulin-immobilized microcarrier was possible, and the promotion activity in the the repeated use was greater than that in the use.
Daniel R. Gossett - One of the best experts on this subject based on the ideXlab platform.
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Label-Free Cell separation and sorting in microfluidic systems
Analytical and Bioanalytical Chemistry, 2010Co-Authors: Daniel R. Gossett, Westbrook M. Weaver, Albert J. Mach, Hamed Amini, Dino Di CarloAbstract:Cell separation and sorting are essential steps in Cell biology research and in many diagnostic and therapeutic methods. Recently, there has been interest in methods which avoid the use of biochemical labels; numerous intrinsic biomarkers have been explored to identify Cells including size, electrical polarizability, and hydrodynamic properties. This review highlights microfluidic techniques used for label-Free discrimination and fractionation of Cell populations. Microfluidic systems have been adopted to precisely handle single Cells and interface with other tools for biochemical analysis. We analyzed many of these techniques, detailing their mode of separation, while concentrating on recent developments and evaluating their prospects for application. Furthermore, this was done from a perspective where inertial effects are considered important and general performance metrics were proposed which would ease comparison of reported technologies. Lastly, we assess the current state of these technologies and suggest directions which may make them more accessible. Figure A wide range of microfluidic technologies have been developed to separate and sort Cells by taking advantage of differences in their intrinsic biophysical properties
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label Free Cell separation and sorting in microfluidic systems
Analytical and Bioanalytical Chemistry, 2010Co-Authors: Daniel R. Gossett, Westbrook M. Weaver, Albert J. Mach, Soojung Claire Hur, Henry T K TseAbstract:Cell separation and sorting are essential steps in Cell biology research and in many diagnostic and therapeutic methods. Recently, there has been interest in methods which avoid the use of biochemical labels; numerous intrinsic biomarkers have been explored to identify Cells including size, electrical polarizability, and hydrodynamic properties. This review highlights microfluidic techniques used for label-Free discrimination and fractionation of Cell populations. Microfluidic systems have been adopted to precisely handle single Cells and interface with other tools for biochemical analysis. We analyzed many of these techniques, detailing their mode of separation, while concentrating on recent developments and evaluating their prospects for application. Furthermore, this was done from a perspective where inertial effects are considered important and general performance metrics were proposed which would ease comparison of reported technologies. Lastly, we assess the current state of these technologies and suggest directions which may make them more accessible.
Henry T K Tse - One of the best experts on this subject based on the ideXlab platform.
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label Free Cell separation and sorting in microfluidic systems
Analytical and Bioanalytical Chemistry, 2010Co-Authors: Daniel R. Gossett, Westbrook M. Weaver, Albert J. Mach, Soojung Claire Hur, Henry T K TseAbstract:Cell separation and sorting are essential steps in Cell biology research and in many diagnostic and therapeutic methods. Recently, there has been interest in methods which avoid the use of biochemical labels; numerous intrinsic biomarkers have been explored to identify Cells including size, electrical polarizability, and hydrodynamic properties. This review highlights microfluidic techniques used for label-Free discrimination and fractionation of Cell populations. Microfluidic systems have been adopted to precisely handle single Cells and interface with other tools for biochemical analysis. We analyzed many of these techniques, detailing their mode of separation, while concentrating on recent developments and evaluating their prospects for application. Furthermore, this was done from a perspective where inertial effects are considered important and general performance metrics were proposed which would ease comparison of reported technologies. Lastly, we assess the current state of these technologies and suggest directions which may make them more accessible.
Ye Fang - One of the best experts on this subject based on the ideXlab platform.
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discovery of β2 adrenoceptor agonists in curcuma zedoaria rosc using label Free Cell phenotypic assay combined with two dimensional liquid chromatography
Journal of Chromatography A, 2018Co-Authors: Ye Fang, Jixia Wang, Yaopeng Zhao, Weijia Zhou, Hongli Jin, Han Zhou, Pengyu Zhang, Yanfang Liu, Xiuli ZhangAbstract:Abstract Traditional Chinese Medicines (TCMs) have been widely used in clinical practice, and provided a rich source for discovering new drug leads. However, efficient identification of active molecules responsible for the therapeutic effects of complex TCMs is still highly challenging. Here, we combined label-Free Cell phenotypic assay with two dimensional liquid chromatography (2DLC) to identify potential β2-adrenoceptor (β2-AR) agonists related to anti-asthmatic effect of Curcuma zedoaria Rosc (C.zedoaria), a commonly used TCM. The ethyl acetate extract of C.zedoaria was first fractionated into 26 fractions. Label-Free Cell phenotypic profiling was then used to locate the active sites. Orthogonal second-dimensional (D2) separation was performed on two fractions displaying agonistic effect at the β2-AR, combined with screening of the D2 fractions to track the activity. Finally, this approach led to the isolation of three known diarylheptanoids, among which diarylheptanoid b exhibited the most potent agonistic activity with an EC50 value of 5.93 μM. This result was further demonstrated through the chemical synthesis of diarylheptanoid b. It is the first time to discover that diarylheptanoids could activate the β2-AR, which may be responsible for the anti-asthmatic effect of C.zedoaria observed traditionally and in clinical application. This study also demonstrates the potential of this integrated strategy for identifying active ingredients and determining the basis of therapeutic materials in complex TCMs.
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Discovery of beta(2)-adrenoceptor agonists in Curcuma zedoaria Rosc using label-Free Cell phenotypic assay combined with two-dimensional liquid chromatography
ELSEVIER SCIENCE BV, 2018Co-Authors: Zhou Weijia, Ye Fang, Wang Jixia, Zhao Yaopeng, Yu Long, Jin Hongli, Zhou Han, Zhang Pengyu, Liu Yanfang, Zhang XiuliAbstract:Traditional Chinese Medicines (TCMs) have been widely used in clinical practice, and provided a rich source for discovering new drug leads. However, efficient identification of active molecules responsible for the therapeutic effects of complex TCMs is still highly challenging. Here, we combined label-Free Cell phenotypic assay with two dimensional liquid chromatography (2DLC) to identify potential beta(2)-adrenoceptor (beta(2)-AR) agonists related to anti-asthmatic effect of Curcuma zedoaria Rosc (C.zedoaria), a commonly used TCM. The ethyl acetate extract of C.zedoaria was first fractionated into 26 fractions. Label Free Cell phenotypic profiling was then used to locate the active sites. Orthogonal second-dimensional (D2) separation was performed on two fractions displaying agonistic effect at the beta(2)-AR, combined with screening of the D2 fractions to track the activity. Finally, this approach led to the isolation of three known diarylheptanoids, among which diarylheptanoid b exhibited the most potent agonistic activity with an EC50 value of 5.93 mu M. This result was further demonstrated through the chemical synthesis of diarylheptanoid b. It is the first time to discover that diarylheptanoids could activate the beta(2)-AR, which may be responsible for the anti -asthmatic effect of C.zedoaria observed traditionally and in clinical application. This study also demonstrates the potential of this integrated strategy for identifying active ingredients and determining the basis of therapeutic materials in complex TCMs. (C) 2018 Elsevier B.V. All rights reserved
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Label-Free Cell Phenotypic Identification of D-Luciferin as an Agonist for GPR35.
Methods in molecular biology (Clifton N.J.), 2016Co-Authors: Huayun Deng, Ye FangAbstract:D-Luciferin (also known as beetle or firefly luciferin) is one of the most widely used bioluminescent reporters for monitoring in vitro or in vivo luciferase activity. The identification of several natural phenols and thieno[3,2-b]thiophene-2-carboxylic acid derivatives as agonists for GPR35, an orphan G protein-coupled receptor, had motivated us to examine the pharmacological activity of D-Luciferin, given that it also contains phenol and carboxylic acid moieties. Here, we describe label-Free Cell phenotypic assays that ascertain D-Luciferin as a partial agonist for GPR35. The agonistic activity of D-Luciferin at the GPR35 shall evoke careful interpretation of biological data when D-Luciferin or its analogues are used as probes.
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Label-Free Cell Phenotypic Assays for Assessing Drug Polypharmacology.
Current pharmaceutical design, 2016Co-Authors: Jixia Wang, Ye Fang, Xiuli Zhang, Xinmiao LiangAbstract:Background: Most drugs exert their biological and physiological effects via binding to protein targets. Although drugs are traditionally optimized against a single protein, most marketed drugs exhibit clinically relevant polypharmacology – the activity of drugs at multiple targets. The wide-spread presence of polypharmacology makes it challenging to assess the mechanisms of action of multi-target drugs. Methods: This paper first reviews approaches for discovering multi-targets of drug molecules, then discusses key characteristics of label-Free Cell phenotypic assays, and finally focuses on how to use these assays to assess drug polypharmacology. Results: labelFree Cell phenotypic assays have ability to provide a holistic view of drug action in living Cells with wide phenotype/ target/pathway coverage, and permit effective deconvolution of the action of multi-target drugs at the whole Cell level. Conclusion: Label-Free Cell phenotypic assays hold great potential in assessing drug polypharmacology.
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Combining label-Free Cell phenotypic profiling with computational approaches for novel drug discovery.
Expert opinion on drug discovery, 2015Co-Authors: Ye FangAbstract:Introduction: Drug discovery is a long and costly process. Innovations and paradigm shifts are essential for continuous improvement in the productivity of pharmaceutical R&D.Areas covered: The author reviews the progress of label-Free Cell phenotypic and computational approaches in early drug discovery since 2004 and proposes a novel paradigm, which combines both approaches.Expert opinion: Label-Free Cell phenotypic profiling techniques offer an unprecedented and integrated approach to comprehend drug–target interactions in their native environments. However, these approaches have disadvantages associated with the lack of molecular details. Computational approaches, including ligand-, structure- and phenotype-based virtual screens, have become versatile tools in the early drug discovery process. However, these approaches mostly predict the binding of drug molecules to targets of interest and are limited to targets that are either well annotated for ligands or that are structurally resolved. Thus, combinin...
