Faculty Profiles - ITO Akira

Genetically engineered angiogenic cell sheets using magnetic force-based gene delivery and tissue fabrication techniques. Reviewed

Hirokazu Akiyama, Akira Ito, Yoshinori Kawabe, Masamichi Kamihira

Biomaterials Vol. 31 ( 6 ) page： 1251-1259 2010

Tissue engineering using magnetite nanoparticles and magnetic force: Heterotypic layers of co-cultured hepatocytes and endothelial cells. Reviewed

Akira Ito, Yohei Takizawa, Hiroyuki Honda, Kenichiro Hata, Hideaki Kagami, Minoru Ueda, Takeshi Kobayashi

Tissue Engineering Vol. 10 ( 5-6 ) page： 833-840 2004

Magnetically triggered transgene expression in mammalian cells by localized cellular heating of magnetic nanoparticles. Reviewed

Akira Ito, Ryoji Teranishi, Kazuki Kamei, Masaki Yamaguchi, Akihiko Ono, Shinya Masumoto, Yuto Sonoda, Masanobu Horie, Yoshinori Kawabe, Masamichi Kamihira

Journal of bioscience and bioengineering Vol. 128 ( 3 ) page： 355 - 364 2019.9

Language：English Publishing type：Research paper (scientific journal)

To develop a remote control system of transgene expression through localized cellular heating of magnetic nanoparticles, a heat-inducible transgene expression system was introduced into mammalian cells. Cells were labeled with magnetic nanoparticles and exposed to an alternating magnetic field. The magnetically labeled cells expressed the transgene in a monolayer and multilayered cell sheets in which cells were heated around the magnetic nanoparticles without an apparent temperature increase in the culture medium. Magnetic cells were also generated by genetically engineering with a ferritin gene, and transgene expression could be induced by exposure to an alternating magnetic field. This approach may be applicable to the development of novel gene therapies in cell-based medicine.

DOI： 10.1016/j.jbiosc.2019.03.008

Neural differentiation of mouse induced pluripotent stem cells using cadherin gene-engineered PA6 feeder cells. Reviewed

Paerwen Paerhati, Akira Ito, Kantaro Yoshioka, Kaori Iwamoto, Sho Fujiwara, Masanobu Horie, Yoshinori Kawabe, Masamichi Kamihira

Journal of bioscience and bioengineering Vol. 127 ( 5 ) page： 633 - 640 2019.5

Language：English Publishing type：Research paper (scientific journal)

Investigating neural differentiation of pluripotent stem cells, including induced pluripotent stem (iPS) cells, is of importance for studying early neural development and providing a potential source of cells for nerve regeneration. Stromal cell-derived inducing activity (SDIA) using PA6 stromal cells promotes neural differentiation of iPS cells. Thus, we hypothesized that cadherin gene-engineered PA6 feeder cells will enhance the performance of SDIA by facilitating cell-cell interactions. Consequently, we created cadherin gene-engineered PA6 cells. Efficiency of neural differentiation from mouse iPS cells on PA6 feeder cells overexpressing E-cadherin gene (46%) or N-cadherin gene (27%) was significantly higher compared with parental PA6 feeder cells (19%). In addition, efficiency of motor neuron differentiation from mouse iPS cells on cadherin-gene engineered feeder cells (E-cadherin, 7.4%; N-cadherin, 11%) was significantly higher compared with parental PA6 feeder cells (4.1%). Altogether, these results indicate that cadherin gene-engineered feeder cells are a potent tool for promoting neural differentiation of pluripotent stem cells.

DOI： 10.1016/j.jbiosc.2018.10.009

Fabricating Muscle-Neuron Constructs with Improved Contractile Force Generation. Reviewed

Arifuzzaman M, Ito A, Ikeda K, Kawabe Y, Kamihira M

Tissue engineering. Part A Vol. 25 ( 7-8 ) page： 563-574 2019.4

Induction of functional tissue-engineered skeletal muscle constructs by defined electrical stimulation. Reviewed

Akira Ito, Yasunori Yamamoto, Masanori Sato, Kazushi Ikeda, Masahiro Yamamoto, Hideaki Fujita, Eiji Nagamori, Yoshinori Kawabe, Masamichi Kamihira

Scientific Reports Vol. 4 page： 4781 2014

Improved recombinant antibody production by CHO cells using a production enhancer DNA element with repeated transgene integration at a predetermined chromosomal site Reviewed

Yoshinori Kawabe, Takanori Inao, Shodai Komatsu, Guan Huang, Akira Ito, Takeshi Omasa, Masamichi Kamihira

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 123 ( 3 ) page： 390 - 397 2017.3

Language：English Publishing type：Research paper (scientific journal) Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN

Chinese hamster ovary (CHO) cells are one of the most useful host cell lines for the production of biopharmaceutical proteins. Although a series of production processes have been refined to improve protein productivity and cost performance, establishing producer cells is still time-consuming and labor-intensive. Recombinase-mediated site-specific gene integration into a predetermined chromosomal locus may enable predictable protein expression, reducing the laborious process of cell screening. We previously developed an accumulative site-specific gene integration system (AGIS) using Cre recombinase and mutated loxP sites for transgene integration and amplification in the CHO cell genome. Epigenetic modifier elements such as insulators are effective DNA cis-regulatory elements for stabilizing transgene expression. Here, we attempted to enhance transgene expression in recombinant CHO cells generated by AGIS using a production enhancer DNA element (PE) derived from the CHO genome. The PE was introduced into an expression unit for a recombinant scFv-Fc antibody. The effect on scFv-Fc productivity of PE position and orientation within the transgene was evaluated, while keeping the background chromosomal structure constant. For the optimal PE arrangement, scFv-Fc productivity was enhanced 2.6-fold compared with an expression unit without a PE. The enhancing effect of the PE on transgene expression was also observed when two or three PE-flanked expression units were inserted as tandem repeats. These results indicate that AGIS using the PE-flanked expression unit is a promising approach for establishing producer cell lines for biopharmaceutical protein production. (C) 2016, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2016.10.011

All-In-One Magnetic Nanoparticles for Thermo-Immunotherapy of Malignant Melanoma

Novel therapy for pancreatic fistula using adipose-derived stem cell sheets treated with mannose. Reviewed

Hirokazu Kaneko, Toshio Kokuryo, Yukihiro Yokoyama, Junpei Yamaguchi, Tokunori Yamamoto, Rei Shibata, Momokazu Gotoh, Toyoaki Murohara, Akira Ito, Masato Nagino

Surgery Vol. 161 ( 6 ) page： 1561–1569 2017

Targeted transgene insertion into the CHO cell genome using Cre recombinase-incorporating integrase-defective retroviral vectors. Reviewed

Yoshinori Kawabe, Takuya Shimomura, Shuohao Huang, Suguru Imanishi, Akira Ito, Masamichi Kamihira

Biotechnology and Bioengineering Vol. 113 ( 7 ) page： 1600-1610 2016

Improved contractile force generation of tissue-engineered skeletal muscle constructs by IGF-I and Bcl-2 gene transfer with electrical pulse stimulation. Reviewed

Kazushi Ikeda, Akira Ito, Masanori Sato, Yoshinori Kawabe, Masamichi Kamihira

Regenerative Therapy Vol. 3 page： 38-44 2016

A new in vitro co-culture model using magnetic force-based nanotechnology. Reviewed

Hiroki Takanari, Keiko Miwa, XianMing Fu, Jun-ichi Nakai, Akira Ito, Kousuke Ino, Hiroyuki Honda, Jun Kunishida, Satoru Konishi, Tobias Opthof, Marcel AG van der Heyden, Itsuo Kodama, Jong-Kook Lee

Journal of Cellular Physiology Vol. 231 ( 10 ) page： 2249-2256 2016

DNA damage-responsive transgene expression mediated by the p53 promoter with transcriptional amplification. Reviewed

Akihiko Ono, Akira Ito, Taiga Suzuki, Masaki Yamaguchi, Yoshinori Kawabe, Masamichi Kamihira

Journal of Bioscience and Bioengineering Vol. 120 ( 4 ) page： 463-466 2015

Effects of type IV collagen on myogenic characteristics of IGF-I gene-engineered myoblast cells. Reviewed

Akira Ito, Masahiro Yamamoto, Kazushi Ikeda, Masanori Sato, Yoshinori Kawabe, Masamichi Kamihira

Journal of Bioscience and Bioengineering. Vol. 119 ( 5 ) page： 596-603 2015

Magnetic-labeled feeder system for mouse pluripotent stem cells. Reviewed

Masanobu Horie, Akira Ito, Takeshi Maki, Yoshinori Kawabe, Masamichi Kamihira

Journal of Bioscience and Bioengineering. Vol. 119 ( 5 ) page： 614-616 2015

Improved transgene integration into the CHO cell genome using the Cre-loxP system. Reviewed

Takanori Inao, Yoshinori Kawabe, Takuro Yamashiro, Yujiro Kameyama, Xue Wang, Akira Ito, Masamichi Kamihira

Journal of Bioscience and Bioengineering. Vol. 120 ( 1 ) page： 99-106 2015

Fabrication of 3D tissue-like structure using magnetite nanoparticles and magnetic force.

Akira Ito, Kosuke Ino, Kazunori Shimizu, Hiroyuki Honda, Masamichi Kamihira

2006 IEEE International Symposium on Micro-Nano Mechanical and Human Science 2006

磁性ナノ粒子を用いた組織工学手法の開発 Invited

井藤　彰、本多裕之

再生医療－日本再生医療学会雑誌 Vol. 5 ( 1 ) page： 84-90 2006

再生医療への応用 Invited

井藤　彰、本多裕之

磁性ビーズのバイオ・環境技術への応用展開 page： 64-74 2006

Magnetic manipulation of a retroviral vector using magnetite cationic liposomes.

Akira Ito, Tetsuya Takahashi, Yujiro Kameyama, Yoshinori Kawabe, Masamichi Kamihira

Proceeding of MHS2008 page： 367-371 2008

Screening of Cytokines to Enhance Vaccine Effects of Heat Shock Protein 70-Rich Tumor Cell Lysate Reviewed

Akira Ito, Masatake Fujioka, Kouji Tanaka, Takeshi Kobayashi, Hiroyuki Honda

Journal of Bioscience and Bioengineering Vol. 100 ( 1 ) page： 36-42 2005

Complete Regression of Hereditary Melanoma in a Mouse Model by Repeated Hyperthermia Using Magnetite Cationic Liposomes Reviewed

Akira Ito, Yoko Nakahara, Masatake Fujioka, Takeshi Kobayashi, Kozue Takeda, Izumi Nakashima, Hiroyuki Honda

Japanese Journal of Hyperthermic Oncology Vol. 21 ( 3 ) page： 179-189 2005

The effect of RGD peptide-conjugated magnetite cationic liposomes on cell growth and cell sheet harvesting Reviewed

Akira Ito, Kousuke Ino, Takeshi Kobayashi, Hiroyuki Honda

Biomaterials Vol. 26 ( 31 ) page： 6185-6193 2005

Ehancement of hepatocyte function through heterotypic cell-cell interactions using E-cadherin-expressing NIH3T3 cells. Reviewed

Akira Ito, Takehiko Kiyohara, Yoshinori Kawabe, Hiroyuki Ijima, Masamichi Kamihira

Proceeding of the 20th JAACT meeting page： 159-164 2008

Heat Immunotherapy Using Magnetic Nanoparticles and Dendritic Cells for T-Lymphoma Reviewed

Kouji Tanaka, Akira ITO, Takeshi Kobayashi, Tatsuyoshi Kawamura, Shinji Shimada, Kazuhiko Matsumoto, Toshiaki Saida, Hiroyuki Honda

Journal of Bioscience and Bioengineering Vol. 100 ( 1 ) page： 112-115 2005

Magnetic Force-Based Mesenchymal Stem Cell Expansion Using Antibody-Conjugated Magnetoliposomes Reviewed

Akira Ito, Eri Hibino, Kazunori Shimizu, Takeshi Kobayashi, Yoichi Yamada, Hideharu Hibi, Minoru Ueda, Hiroyuki Honda

Journal of Biomedical Materials Research Vol. 75 ( 2 ) page： 320-327 2005

Intratumoral Injection of Immature Dendritic Cells Enhances Antitumor Effect of Hyperthermia Using Magnetic Nanoparticles Reviewed

Kouji Tanaka, Akira Ito, Takeshi Kobayashi, Tatsuyoshi Kawamura, Shinji Shimada, Kazuhiko Matsumoto, Toshiaki Saida, Hiroyuki Honda

International Journal of Cancer Vol. 116 ( 4 ) page： 624-633 2005

Anticancer effect of hyperthermia on prostate cancer mediated by magnetite cationic liposomes and immune-response induction in transplanted syngeneic rats. Reviewed

Noriyasu Kawai, Akira Ito, Yoko Nakahara, Atsuya Hikosaka, Mitsuru Hutakuchi, Tomoyuki Shirai, Hiroyuki Honda, Takeshi Kobayashi, Kenjiro Kohri

The Prostate Vol. 64 ( 4 ) page： 373-381 2005

A Mechanism of Antitumor Immunity Induced by Hyperthermia Invited Reviewed

Akira Ito, Takeshi Kobayashi, Hiroyuki Honda

Japanese Journal of Hyperthermic Oncology Vol. 21 ( 1 ) page： 1-11 2005

Heat induction of reporter gene expression via the gadd 153 promoter and its possible application to hyperthermia treatment of cancer. Reviewed

Isabelle Anne Bouhon, Akira Ito, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Cytotechnology Vol. 33 ( 1 ) page： 131-137 2000

磁性ナノ粒子を用いた管状組織の構築

伊野浩介、井藤彰、林田真生、小林猛、松沼寛、各務秀明、上田実、本多裕之

化学工学シンポジウムシリーズ「診断・治療システムと化学工学」 Vol. 79 page： 63-67 2005

抗体結合型マグネトリポソームを用いた間葉系幹細胞の濃縮培養法

清水一憲、井藤彰、日比野恵里、小林猛、山田陽一、日比英晴、上田実、本多裕之

化学工学シンポジウムシリーズ「診断・治療システムと化学工学」 Vol. 79 page： 68-72 2005

磁性ナノ粒子を利用した再生医療技術"Mag-TE"の開発 Invited

井藤　彰

東海化学工業会会報 Vol. 247 page： 14-16 2005

磁性ビーズの医療への応用

井藤　彰、本多裕之

日本応用磁気学会　第141回研究会資料 page： 9-14 2005

ナノ磁性微粒子を用いた生体組織構築 Invited

井藤　彰、本多裕之

ケミカルエンジニヤリング Vol. 50 ( 5 ) page： 13-18 2005

メラノーマに対するHeat Immunotherapyの開発 Invited

井藤　彰、小林　猛、本多裕之

Vol. 19 ( 3 ) page： 298-305 2005

免疫誘導型ハイパーサイミア“Heat Immunotherapy”の開発 Invited

井藤　彰、小林　猛、本多裕之

癌の臨床 Vol. 50 ( 13 ) page： 1127-1131 2005

Functional magnetic particles for medical application. Invited Reviewed

Shinkai M, Ito A.

Adv Biochem Eng Biotechnol Vol. 91 page： 191-220 2004

Local hyperthermia enhances thrombosis in aneurysms containing platinum coils Reviewed

Suzuki O, Miyachi S, Okamoto T, Ito A, Shinkai M, Honda H, Kobayashi T, Negoro M, Yoshida J

Interventional Neuroradiology Vol. 10 ( 3 ) page： 203-211 2004

Introduction of TNF-alfa gene expression by heat inducible promoter gadd153. Reviewed

Akira Ito, Masashige Shinkai, Isabelle Anne Bouhon, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Japanese Journal of Hyperthermic Oncology Vol. 16 ( 2 ) page： 91-98 2000

Tissue engineering using magnetite nanoparticles and magnetic force: heterotypic layers of co-cultured hepatocytes and endothelial cells. Reviewed

Akira Ito, Yohei Takizawa, Hiroyuki Honda, Ken-ichiro Hata, Hideaki Kagami, Minoru Ueda and Takeshi Kobayashi

Tissue Engineering Vol. 10 ( 5/6 ) page： 833-840 2004

A new methodology of mesenchymal stem cell expansion using magnetic nanoparticles Reviewed

Akira Ito, Eri Hibino, Hiroyuki Honda, Ken-ichiro Hata, Hideaki Kagami, Minoru Ueda and Takeshi Kobayashi

Biochemical Engineering Journal Vol. 20 page： 119-125 2004

Radiation-inducible TNF-alpha gene expression under stress-inducible promoter gadd153 for cancer therapy. Reviewed

Akira Ito, Masashige Shinkai, Kazumi Hakamada, Hiroyuki Honda, Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 92 ( 6 ) page： 598-601 2001

磁性微粒子を用いた治療技術開発 Invited

井藤　彰、本多裕之、小林　猛

バイオインダストリー Vol. 21 ( 8 ) page： 48-54 2004

磁性微粒子を用いたTissue Engineeringの開発 Invited

井藤　彰、本多裕之

ケミカルエンジニアリング Vol. 49 ( 6 ) page： 6-12 2004

磁性ナノ微粒子を用いたTissue Engineering –Mag-TEの開発- Invited

井藤　彰、本多裕之、小林　猛

化学工学会バイオ部会Newsletter Vol. 10 ( 3 ) page： 1 2004

Construction of Multilayerd Cell Sheets Using Magnetite Nanoparticles and Magnetic Force

Akira Ito, Hiroyuki Honda, Ken-ichiro Hata, Hideaki Kagami, Minoru Ueda, Takeshi Kobayashi

Proceding of YABEC'04 page： 142 2004

Skeletal Muscle Tissue Engineering Using Functional Magnetite Nanoparticles.

Akira Ito, Hirokazu Akiyama, Yasunori Yamamoto, Yoshinori Kawabe, Masamichi Kamihira

Proceeding of MHS2009 page： 379-382 2009

Heat shock protein 70 gene therapy combined with hyperthermia using magnetic nanoparticles Reviewed

Akira Ito, Fumiko Matsuoka, Hiroyuki Honda and Takeshi Kobayashi

Cancer Gene Therapy Vol. 10 ( 12 ) page： 918-925 2003

Heat shock protein 70 expression induces an antitumor immunity during intracellular hyperthermia using magnetite nanoparticles Reviewed

Akira Ito, Masashige Shinkai, Hiroyuki Honda, Kazuhiro Yoshikawa, Shinsuke Saga, Toshihiko Wakabayashi, Jun Yoshida and Takeshi Kobayashi

Cancer Immunology Immunotherapy Vol. 52 ( 2 ) page： 80-88 2003

Transglutaminase-mediated gelatin matrices incorporating cell adhesion factors as a biomaterial for tissue engineering Reviewed

Akira Ito, Atsunori Mase, Yohei Takizawa, Masashige Shinkai, Hiroyuki Honda, Ken-ichiro Hata, Minoru Ueda and Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 95 ( 2 ) page： 196-199 2003

Tumor regression by combined immunotherapy and hyperthermia using magnetic nanoparticles in an experimental subcutaneous murine melanoma Reviewed

Akira Ito, Kouji Tanaka, Kazuyoshi Kondo, Masashige Shinkai, Hiroyuki Honda, Kazuhiko Matsumoto, Toshiaki Saida and Takeshi Kobayashi

Cancer Science Vol. 94 ( 3 ) page： 308-313 2003

Proliferation and stratification of keratinocyte on cultured amniotic epithelial cells for tissue engineering Reviewed

Akira Ito, Yohei Takizawa, Masashige Shinkai, Hiroyuki Honda, Ken-Ichiro Hata, Minoru Ueda, Naohiko Kuno, Atsuo Itakura and Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 95 ( 6 ) page： 589-593 2003

Complete regression of mouse mammary carcinoma with a size greater than 15 mm by frequent repeated hyperthermia using magnetic nanoparticles Reviewed

Akira Ito, Kouji Tanaka, Hiroyuki Honda, Shigeru Abe, Hideyo Yamaguchi and Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 96 ( 4 ) page： 360-363 2003

Time course of biodistribution and heat generation of magnetite cationic liposomes in mouse model. Reviewed

Akira Ito, Yoko Nakahara, Kouji Tanaka, Yuko Kuga, Hiroyuki Honda and Takeshi Kobayashi

Japanese Journal of Hyperthermic Oncology Vol. 19 ( 3 ) page： 151-159 2003

Selective hyperthermia using magnetoliposomes to target cervical lymph node metastasis in a rabbit tongue tumor model. Reviewed

Shigeaki Hamaguchi, Iwai Tohnai, Akira Ito, Kenji Mitsudo, Toshio Shigetomi, Masafumi Ito, Hiroyuki Honda, Takeshi Kobayashi and Minoru Ueda

Cancer Science Vol. 94 ( 9 ) page： 834-839 2003

磁性微粒子を活用した特異的な診断・治療システム Invited

井藤　彰、本多裕之、小林　猛

化学工学 Vol. 67 ( 12 ) page： 692-695 2003

Transglutaminase-mediated gelatin matrices incorporating cell adhesion factors as a biomaterial for tissue engineering. Reviewed

Akira Ito, Atsuhiko Mase, Yohei Takizawa, Masashige Shinkai, Hiroyuki Honda, Kenichiro Hata, Minoru Ueda, Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 95 ( 2 ) page： 196-199 2003

Bystander-killing effect and cyclic induction of TNF-alfa gene under heat inducible promoter gadd153. Reviewed

Akira Ito, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 90 ( 4 ) page： 437-441 2000

Heat-inducible TNF-alpha gene therapy combined with hyperthermia using magnetic nanoparticles as a novel tumor-targeted therapy. Reviewed

Akira Ito, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Cancer Gene Therapy Vol. 8 ( 9 ) page： 649-654 2001

Effective solitary hyperthermia treatment of malignant glioma using stick type CMC-magnetite. In vivo study Reviewed

Ohno T, Wakabayashi T, Takemura A, Yoshida J, Ito A, Shinkai M, Honda H and Kobayashi T

Journal of Neuro-Oncology Vol. 56 ( 3 ) page： 233 2002

バイオターゲティングのための生体分子デザイン

井藤　彰、本多裕之、小林　猛

未来材料 Vol. 2 ( 8 ) page： 5 2002

リポソームを用いた腫瘍への薬剤ターゲティング

井藤　彰、本多裕之、小林　猛

Vol. 17 ( 4 ) page： 347 2002

Radiation-inducible TNF-a gene expression under stress-inducible promoter gadd153 for cancer therapy Reviewed

Akira Ito, Masashige Shinkai, Kazumi Hakamada, Hiroyuki Honda, Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 92 ( 6 ) page： 598 2001

Heat-inducible TNF-alpha gene therapy combined with hyperthermia using magnetic nanoparticles as a novel tumor-targeted therapy Reviewed

Akira Ito, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Cancer Gene Therapy Vol. 8 ( 9 ) page： 649 2001

Augmentation of MHC class I antigen presentation via heat shock protein expression by hyperthermia Reviewed

Akira Ito, Masashige Shinkai, Hiroyuki Honda, Toshihiko Wakabayashi, Jun Yoshida, Takeshi Kobayashi

Cancer Immunology Immunotherapy Vol. 50 ( 10 ) page： 515 2001

Bystander-killing effect and cyclic induction of TNF-α gene under heat inducible promoter gadd153 Reviewed

Akira Ito, Masashige Shinkai, Isabelle Anne Bouhon, Hiroyuki Honda, Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 90 ( 4 ) page： 437 2000

Heat induction of reporter gene expression via the gadd 153 promoter and its possible application to hyperthermia treatment of cancer Reviewed

Isabelle Anne Bouhon, Akira Ito, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Cytotechnology Vol. 33 ( 1 ) page： 131 2000

Introduction of TNF-α gene expression by heat inducible promoter gadd153 Reviewed

Akira Ito, Masashige Shinkai, Isabelle Anne Bouhon, Hiroyuki Honda, Takeshi Kobayashi

Japanese Journal of Hyperthermic Oncology Vol. 16 ( 2 ) page： 91 2000

Effective solitary hyperthermia treatment of malignant glioma using stick type CMC-magnetite. In vivo study. Reviewed

Takanari Ohno, Toshihiko Wakabayashi, Atsuhito Takemura, Jun Yoshida, Akira Ito, Masashige Shinkai, Hiroyuki Honda, Takeshi Kobayashi

Journal of Neuro-Oncology Vol. 56 ( 5 ) page： 233-239 2000

Complete regression of mouse mammary carcinoma with a size greater than 15 mm by frequent repeated hyperthermia using magnetic nanoparticles. Reviewed

Akira Ito, Koji Tanaka, Hiroyuki Honda, Shigeru Abe, Hideyo Yamaguchi, Takeshi Kobayashi

Journal of Bioscience and Bioengineering Vol. 96 ( 4 ) page： 360-363 2003

ガン治療を目的とした遺伝子発現の誘導

新海政重、井藤　彰、イザベル　ブホン、本多裕之、小林　猛

ケミカルエンジニヤリング Vol. 44 page： 9 1999

Kaneko, M; Saito, R; Nishikawa, A; Ito, A

ADVANCED HEALTHCARE MATERIALS page： e2500260 2025.3

Fabrication of phospholipid polymer-modified alginate hydrogels for bioartificial pancreas. Reviewed

Ryo Futatsubashi, Masahiro Kaneko, Akira Ito.

Journal of Bioscience and Bioengineering Vol. 139 ( 3 ) page： 242 - 248 2025.3

Amphiphilic phospholipid polymers as a cryoprotectant for vitrification and nanowarming of rat livers

Kaneko, M; Takizawa, N; Wakabayashi, T; Kaneoka, H; Ito, A

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 139 ( 1 ) page： 70 - 78 2025.1

Language：English Publisher：Journal of Bioscience and Bioengineering

Liver biobanking is a promising approach that saves the lives of patients with end-stage liver disease. Cryopreservation based on vitrification enables semi-permanent organ preservation, contributing to overcome the shortage of donors for liver transplants. A technical challenge in cryopreservation of transplantable organs lies in thawing methodology, and conventional convective warming cannot maintain the glassy state during thawing because of the large temperature gradient between the inner and outer parts of the organs, leading to ice formation and damage of cells in the organ. Nanowarming, in which magnetic nanoparticles are dispersed in a vitrification solution and heated by exposure of alternating magnetic field, can achieve uniform and rapid heating of organs. Herein, we report that amphiphilic phospholipid polymers composed of 2-methacryloyloxyethyl phosphorylcholine and n-butyl methacrylate can function as a cryoprotectant for nanowarming. The amphiphilic phospholipid polymers enhanced the viability of primary rat hepatocytes after vitrification. Moreover, the polymers enhanced the dispersion stability of magnetic nanoparticles in vitrification solution, and the perfusion of the vitrification solution with magnetic nanoparticles into rat livers through portal vein provided uniform distribution of the nanoparticles in the liver. After perfusion, the vitrified liver was successfully thawed rapidly and uniformly by nanowarming, which maintained tissue integrity and cell viability.

DOI： 10.1016/j.jbiosc.2024.10.003

Bone morphogenetic protein signaling inhibitor improves differentiation and function of 3D muscle construct fabricated using C2C12

Ding, R; Xi, Y; Ito, A; Shimizu, K; Nagamori, E; Fujita, H; Kawamoto, T; Horie, M

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 137 ( 6 ) page： 480 - 486 2024.6

Language：English Publisher：Journal of Bioscience and Bioengineering

Functional tissue-engineered artificial skeletal muscle tissue has great potential for pharmacological and academic applications. This study demonstrates an in vitro tissue engineering system to construct functional artificial skeletal muscle tissues using self-organization and signal inhibitors. To induce efficient self-organization, we optimized the substrate stiffness and extracellular matrix (ECM) coatings. We modified the tissue morphology to be ring-shaped under optimized self-organization conditions. A bone morphogenetic protein (BMP) inhibitor was added to improve overall myogenic differentiation. This supplementation enhanced the myogenic differentiation ratio and myotube hypertrophy in two-dimensional cell cultures. Finally, we found that myotube hypertrophy was enhanced by a combination of self-organization with ring-shaped tissue and a BMP inhibitor. BMP inhibitor treatment significantly improved myogenic marker expression and contractile force generation in the self-organized tissue. These observations indicated that this procedure may provide a novel and functional artificial skeletal muscle for pharmacological studies.

DOI： 10.1016/j.jbiosc.2024.03.005

Transplantable cell-encapsulation device using a semipermeable ethylene-vinyl alcohol copolymer membrane in a mouse diabetic model

Bone morphogenic protein signaling inhibitor improves differentiation and function of 3D muscle construct fabricated using C2C12 Reviewed

Ran Ding, Yuan Xi, Akira Ito, Kazunori Shimizu, Eiji Nagamori, Hideaki Fujita, Takuo Kawamoto, Masanobu Horie

Journal of Bioscience and Bioengineering Vol. 137 ( 6 ) page： 480 - 486 2024

Kaneko, M; Moriguchi, H; Futatsubashi, R; Ayano, S; Kobayashi, G; Ito, A

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 136 ( 5 ) page： 415 - 422 2023.11

Language：English Publisher：Journal of Bioscience and Bioengineering

Cell-based therapy is an attractive approach, and encapsulation of therapeutic cells is a promising strategy because it prevents immune responses and allows transplanted cells to be retrieved in case of dysfunction. Bioartificial pancreas, in which insulin-secreting cells are encapsulated in a semipermeable membrane bag, is a new class of medical device for treating type-I diabetes. In this study, we developed a macroencapsulation device in which the pancreatic beta cell line MIN6 was encapsulated in a semipermeable bag made of an ethylene-vinyl alcohol copolymer membrane. In vitro evaluation of ATP and insulin levels revealed that MIN6 cells grown in Matrigel within the device secreted insulin in response to glucose levels. Transplantation of the device lowered blood glucose levels for 30 days in diabetic mice. Histological observation revealed that MIN6 cells formed spheroids in Matrigel, and no host cells were detected within the device. Blood levels of inflammatory cytokines in the transplanted mice were similar to those in non-transplanted mice, and antibody levels in the device were lower than those in the intraperitoneal fluid. These results suggest that the semipermeable ethylene-vinyl alcohol copolymer membrane developed in this study is useful for cell encapsulation in cell-based therapies, including beta-cell macroencapsulation for type-1 diabetes.

DOI： 10.1016/j.jbiosc.2023.09.001

Effective magnetic hyperthermia induced by mitochondria-targeted nanoparticles modified with triphenylphosphonium-containing phospholipid polymers. Reviewed

A transplantable cell-encapsulation device using a semipermeable ethylene-vinyl alcohol membrane in a mouse diabetic model Reviewed

Masahiro Kaneko, Hiroaki Moriguchi, Ryo Futatsubashi, Satoru Ayano, Goro Kobayashi, Akira Ito

2023.9

機能性磁性ナノ粒子を用いた再生医療プロセスの開発 Invited

井藤　彰

セラミックス Vol. 58 ( 9 ) page： 584 - 587 2023.9

Expansion of human mesenchymal stem cells on poly(vinyl alcohol) microcarriers Reviewed

Masahiro Kaneko, Airi Sato, Satoru Ayano, Akio Fujita, Goro Kobayashi, Akira Ito

2023.8

Masahiro Kaneko, Hiroto Yamazaki, Takahiro Ono, Masanobu Horie, Akira Ito

Cancer Science Vol. 114 page： 3750 - 3758 2023.7

Nanowarming of vitrified pancreatic islets as a cryopreservation technology for transplantation

Wakabayashi, T; Kaneko, M; Nakai, T; Horie, M; Fujimoto, H; Takahashi, M; Tanoue, S; Ito, A

BIOENGINEERING & TRANSLATIONAL MEDICINE Vol. 8 ( 4 ) page： e10416 2023.7

Language：English Publisher：Bioengineering and Translational Medicine

Biobanking of pancreatic islets for transplantation could solve the shortage of donors, and cryopreservation of vitrified islets is a possible approach. However, a technological barrier is rewarming of large volumes both uniformly and rapidly to prevent ice formation due to devitrification. Here, we describe successful recovery of islets from the vitrified state using a volumetric rewarming technology called “nanowarming,” which is inductive heating of magnetic nanoparticles under an alternating magnetic field. Convective warming using a 37°C water bath as the gold standard for rewarming of vitrified samples resulted in a decrease in the viability of mouse islets in large volumes (>1 ml) owing to devitrification caused by slow warming. Nanowarming showed uniform and rapid rewarming of vitrified islets in large volumes. The viability of nanowarmed islets was significantly improved and islets transplanted into streptozotocin-induced diabetic mice successfully lowered serum glucose. The results suggest that nanowarming will lead to a breakthrough in biobanking of islets for transplantation.

DOI： 10.1002/btm2.10416

Combination of magnetic hyperthermia and immunomodulators to drive complete tumor regression of poorly immunogenic melanoma Open Access

Nishikawa, A; Suzuki, Y; Kaneko, M; Ito, A

CANCER IMMUNOLOGY IMMUNOTHERAPY Vol. 72 ( 6 ) page： 1493 - 1504 2023.6

Language：English Publisher：Cancer Immunology, Immunotherapy

Hyperthermia using magnetic nanoparticles enables tumor-specific heating and can destroy tumor tissues. This approach works as in situ vaccination with tumor antigens released from dying tumor cells. However, in situ vaccination caused by magnetic hyperthermia is often insufficient to induce complete regression of poorly immunogenic tumors surrounded by an immunosuppressive microenvironment. In this study, we explored a novel strategy for immunotherapy using magnetic hyperthermia to regress poorly immunogenic melanoma. Magnetic hyperthermia induced tumor cell death in a B16-F10 melanoma mouse model. After hyperthermia treatment, we found elevated levels of HMGB1, which is known to be released from dying cells to promote inflammation, and the proinflammatory cytokine TNF-α was increased in serum of the mice. Systemic administration of glycyrrhizin, an HMGB1 inhibitor, reduced the levels of TNF-α in serum and successfully delayed the regrowth of tumors after magnetic hyperthermia. To achieve complete tumor regression, TLR9 activation by intratumor injection of CpG was combined with systemic administration of anti-PD-1 antibody and anti-CTLA-4 antibody. The combination therapy of magnetic hyperthermia at 46°C with the immunomodulators (glycyrrhizin+CpG+anti-PD-1+anti-CTLA-4) achieved complete tumor regression in 80% of growing 5-mm B16-F10 tumors. These findings have important implications for the development of novel cancer immunotherapy using magnetic hyperthermia for poorly immunogenic tumors.

DOI： 10.1007/s00262-022-03345-8

A Sulfur Containing Melanogenesis Substrate, N-Pr-4-S-CAP as a Potential Source for Selective Chemoimmunotherapy of Malignant Melanoma Invited Reviewed

Yasuaki Tamura, Akira Ito, Kazumasa Wakamatsu, Toshihiko Torigoe, Hiroyuki Honda, Shosuke Ito, Kowichi Jimbow

International Journal of Molecular Sciences Vol. 24 ( 6 ) page： 5235 2023.3

A Sulfur Containing Melanogenesis Substrate, N-Pr-4-S-CAP as a Potential Source for Selective Chemoimmunotherapy of Malignant Melanoma International journal Open Access

Yasuaki Tamura, Akira Ito, Kazumasa Wakamatsu, Toshihiko Torigoe, Hiroyuki Honda, Shosuke Ito, Kowichi Jimbow

International Journal of Molecular Sciences Vol. 24 ( 6 ) 2023.3

Language：English Publishing type：Research paper (scientific journal)

N-propionyl-4-S-cysteaminylphenol (N-Pr-4-S-CAP) is a substrate for tyrosinase, which is a melanin biosynthesis enzyme and has been shown to be selectively incorporated into melanoma cells. It was found to cause selective cytotoxicity against melanocytes and melanoma cells after selective incorporation, resulting in the induction of anti-melanoma immunity. However, the underlying mechanisms for the induction of anti-melanoma immunity remain unclear. This study aimed to elucidate the cellular mechanism for the induction of anti-melanoma immunity and clarify whether N-Pr-4-S-CAP administration could be a new immunotherapeutic approach against melanoma, including local recurrence and distant metastasis. A T cell depletion assay was used for the identification of the effector cells responsible for N-Pr-4-S-CAP-mediated anti-melanoma immunity. A cross-presentation assay was carried out by using N-Pr-4-S-CAP-treated B16-OVA melanoma-loaded bone marrow-derived dendritic cells (BMDCs) and OVA-specific T cells. Administration of N-Pr-4-S-CAP induced CD8+ T cell-dependent anti-melanoma immunity and inhibited the growth of challenged B16F1 melanoma cells, indicating that the administration of N-Pr-4-S-CAP can be a prophylactic therapy against recurrence and metastasis of melanoma. Moreover, intratumoral injection of N-Pr-4-S-CAP in combination with BMDCs augmented the tumor growth inhibition when compared with administration of N-Pr-4-S-CAP alone. BMDCs cross-presented a melanoma-specific antigen to CD8+ T cells through N-Pr-4-S-CAP-mediated melanoma cell death. Combination therapy using N-Pr-4-S-CAP and BMDCs elicited a superior anti-melanoma effect. These results suggest that the administration of N-Pr-4-S-CAP could be a new strategy for the prevention of local recurrence and distant metastasis of melanoma.

DOI： 10.3390/ijms24065235

Novel nano-thermal ablation therapy using functionalized heat-generating nanoparticles for solid cancer treatment Invited Reviewed

Tomio Morino, Noriyasu Kawai, Akira Ito, Takeshi Kobayashi, Takahiro Yasui

World Journal of Cancer and Oncology Research Vol. 2 ( 1 ) page： 29 - 46 2023.2

Molecular events in the melanogenesis cascade as novel melanoma-targeted small molecules: Principle and development Invited Reviewed

Kazumasa Wakamatsu, Akira Ito, Yasuaki Tamura, Tokimasa Hida, Takafumi Kamiya, Toshihiko Torigoe, Hiroyuki Honda, Shosuke Ito, Kowichi Jimbow

Cancers Vol. 14 ( 22 ) page： 5588 2022.11

ハイパーサーミアにおける酸化鉄ナノフラワーの開発－ナノ構造と発熱能の関係－ Invited Reviewed

金子真大，井藤　彰

Vol. 38 ( 3 ) page： 63 - 64 2022.10

Induction of immunogenic cell death in murine colon cancer cells by ferrocene‐containing redox phospholipid polymers Reviewed

Masahiro Kaneko, Akio Yamaguchi, Akira Ito

Cancer Science Vol. 113 ( 10 ) page： 3558 - 3565 2022.10

Nanowarming of vitrified pancreatic islets as a cryopreservation technology for transplantation Reviewed

Taisei Wakabayashi, Masahiro Kaneko, Tomoki Nakai, Masanobu Horie, Hiroyuki Fujimoto, Maazumi Takahashi, Shota Tanoue, Akira Ito

Bioengineering & Translational Medicine 2022.9

Immunomodulation of melanoma by chemo-thermo-immunotherapy using conjugates of melanogenesis Substrate NPrCAP and magnetite nanoparticles: a review Invited Reviewed

Yasuaki Tamura, Akira Ito, Kazumasa Wakamatsu, Takafumi Kamiya, Toshihiko Torigoe, Hiroyuki Honda, Toshiharu Yamashita, Hisashi Uhara, Shosuke Ito, Kowichi Jimbow

International Journal of Molecular Sciences Vol. 23 ( 12 ) page： 6457 2022.6

Hypoxia-responsive expression of vascular endothelial growth factor for induction of angiogenesis in artificial three-dimensional tissues. Reviewed

Shinya Masumoto, Akihiko Ono, Akira Ito, Yoshinori Kawabe, Masamichi Kamihira

Journal of Bioscience and Bioengineering Vol. 132 ( 4 ) page： 399 - 407 2021.10

Contractile activity of myotubes derived from human induced pluripotent stem cells: A model of duchenne muscular dystrophy Reviewed International journal Open Access

Kantaro Yoshioka, Akira Ito, Masanobu Horie, Kazushi Ikeda, Sho Kataoka, Keiichiro Sato, Taichi Yoshigai, Hidetoshi Sakurai, Akitsu Hotta, Yoshinori Kawabe, Masamichi Kamihira

Cells Vol. 10 ( 10 ) page： 2556 2021.10

Language：English Publishing type：Research paper (scientific journal)

Duchenne muscular dystrophy (DMD) is a genetic disorder that results from deficiency of the dystrophin protein. In recent years, DMD pathological models have been created using induced pluripotent stem (iPS) cells derived from DMD patients. In addition, gene therapy using CRISPR-Cas9 technology to repair the dystrophin gene has been proposed as a new treatment method for DMD. However, it is not known whether the contractile function of myotubes derived from gene-repaired iPS cells can be restored. We therefore investigated the maturation of myotubes in electrical pulse stimulation culture and examined the effect of gene repair by observing the contractile behaviour of myotubes. The contraction activity of myotubes derived from dystrophin-gene repaired iPS cells was improved by electrical pulse stimulation culture. The iPS cell method used in this study for evaluating muscle contractile activity is a useful technique for analysing the mechanism of hereditary muscular disease pathogenesis and for evaluating the efficacy of new drugs and gene therapy.

DOI： 10.3390/cells10102556

Development of a genetically modified hepatoma cell line with heat-inducible high liver function. Reviewed International journal Open Access

Hiroyuki Kitano, Yuki Nagae, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira

Cytotechnology Vol. 73 ( 3 ) page： 353 - 362 2021.6

Language：English Publishing type：Research paper (scientific journal)

Hepatoma cells are a promising cell source for the construction of bioartificial liver (BAL) systems owing to their high proliferative capability. However, their low liver function compared with primary hepatocytes is a major problem. In a previous study, we established a genetically modified hepatoma cell line, Hepa/8F5, in which eight liver-enriched transcription factor (LETF) genes were transduced into mouse hepatoma Hepa1-6 cells using a drug-inducible transactivator system. These cells proliferate actively under normal culture conditions, meaning that large quantities can be prepared easily. When the overexpression of the LETFs is induced by the addition of an inducer drug, cell growth stops and cell morphology changes with concomitant high expression of liver functions. However, the liver functions largely depend on the presence of the inducer drug, which must be continuously added to maintain these enhanced functions. In the present study, we attempted to modify the method of induction of LETF overexpression in Hepa/8F5 cells to remove the requirement for continual drug addition. To this end, we constructed a system in which the artificial transactivator was transcribed and amplified under the control of a heat-shock protein promoter, and introduced the system into the genome of Hepa/8F5 cells. In our modified cell line, heat-triggered LETF expression was confirmed to induce high liver function. After drug-screening of transfected cells, we established a hepatoma cell line (Hepa/HS), which exhibited high, heat-inducible liver functions. The Hepa/HS cells may represent a new cell source for hepatic studies such as the construction of BAL systems. Supplementary Information: The online version of this article (10.1007/s10616-021-00457-4) contains supplementary material, which is available to authorized users.

DOI： 10.1007/s10616-021-00457-4

Miniaturized skeletal muscle tissue fabrication for measuring contractile activity Reviewed

Journal of Bioscience and Bioengineering Vol. 131 ( 4 ) page： 434 - 441 2021.4

Magnetically triggered transgene expression system using magnetic nanoparticles Invited Reviewed

Akira Ito

Drug Delivery System 2021

LINE-1 vectors mediate recombinant antibody gene transfer by retrotransposition in Chinese hamster ovary cells. Reviewed

Feiyang Zheng, Yoshinori Kawabe, Mai Murakami, Mamika Takahashi, Kyoka Nishihata, Souichiro Yoshida, Akira Ito, Masamichi Kamihira

Biotechnology Journal Vol. 16 ( 7 ) page： 2000620 2021

Intramyocardial Transplantation ofHuman iPS Cell–Derived CardiacSpheroids Improves Cardiac Function inHeart Failure Animals. Reviewed

JACC: Basic to Translational Science Vol. 36 ( 4 ) page： 101 - 108 2021

Heat dose based large tumor treatment with multiple site injections of heat-generating nanoparticles dispersible within tumor tissue Reviewed

2. Tomio Morino, Akira Ito, Toshiki Etani, Taku Naiki, Noriyasu Kawai, Takahiro Yasui

Thermal Medicine Vol. 36 ( 4 ) page： 104 - 108 2020.12

Heat Dose Based Large Tumor Treatment with Multiple Site Injections of Heat-generating Nanoparticles Dispersible within Tumor Tissue

MORINO TOMIO, ITO AKIRA, ETANI TOSHIKI, NAIKI TAKU, KAWAI NORIYASU, YASUI TAKAHIRO

Thermal Medicine Vol. 36 ( 4 ) page： 101 - 108 2020.12

Language：English Publisher：Japanese Society for Thermal Medicine

Solid cancer therapy based on necrosis induction with heat-generating nanoparticles has been developed in Japan. Heat was induced from intratumorally injected magnetite cationic lipid composite particles (MCL particles) by alternating magnetic field irradiations to kill cancer cells nearby located. In our previous report we have showed importance of heat dose index in vivo (J/cm3 tumor volume) for tumor regression when 45 mg MCL particles were injected at a single site of 1.36 cm3 tumor. Purpose of this study is to show rationale and utility of multiple site injections of MCL particles for treatment of large tumors more than 1.36 cm3.Rat mammary tumors were induced by 7,12-dimethylbenz［a］anthracene (DMBA) and tumors in range of 2.19 ～ 3.81 cm3 were applied to treatment experiment. Treatment condition was designed to reproduce the heat generation condition of the 1.36 cm3 tumor treatment at every multiple injection site in rat mammary tumors. Tumor volume divided by number of injection sites (cm3/site) was set to close to 1.36 cm3/site and 45 mg MCL particles were administered at multiple sites to keep even spaces among injection sites. Three irradiation conditions were set to give close heat dose in vivo (J/cm3) of the 1.36 cm3 tumor treatment. Treatment under designed conditions resulted in complete regression of rat tumors at 21 days after the treatment, showing theoretical validity of design procedures for the multiple site injections. Novel concept of necrosed tumor volume from an injection site (cm3/site) and its actual value under a standard injection condition of 45 mg-MCL/site were described and its use in clinic was discussed.

DOI： 10.3191/thermalmed.36.101

CiNii Research

ハイパーサーミアの新展開 -基礎医学・臨床応用に関する最近の進歩- Reviewed

近藤隆、鈴木文男、水上達治、斎藤淳一、河合憲康、井藤　彰

放射線生物研究 Vol. 55 ( 3 ) page： 210 - 228 2020.9

Magnetic heating of nanoparticles as a scalable cryopreservation technology for human induced pluripotent stem cells

Ito Akira, Yoshioka Kantaro, Masumoto Shinya, Sato Keiichiro, Hatae Yuki, Nakai Tomoki, Yamazaki Takashi, Takahashi Masazumi, Tanoue Shota, Horie Masanobu

SCIENTIFIC REPORTS Vol. 10 ( 1 ) 2020.8

Heat Dose Index in vivo for Cancer Therapy Using Heat-generating Nanoparticles Named Magnetite Cationic Liposomes and Alternating Magnetic Field Irradiator

MORINO TOMIO, TANOUE SHOTA, MIYATA SHUICHIRO, HIRAYAMA KOTARO, ITO AKIRA, ETANI TOSHIKI, NAIKI TAKU, KAWAI NORIYASU, YASUI TAKAHIRO

Thermal Medicine Vol. 36 ( 2 ) page： 47 - 58 2020.7

Language：English Publisher：Japanese Society for Thermal Medicine

Cancer clinical research using heat-generating nanoparticles named magnetite cationic liposomes (MCL) and alternating magnetic field (AMF) irradiator has been conducted. Heat generation from intratumorally injected MCL particles was triggered by AMF irradiation to kill cancer cells nearby located. Tumor temperature was monitored as index to control treatment condition but efficacy was variable from complete regression to ineffective. In order to improve efficacy, we have proposed novel index of heat dose in vivo (J/cm3 tumor volume). Purpose of this study was to reveal actual heat dose in vivo and discuss its utility as index.In order to enable to estimate heat dose, heat generation activity of MCL particles (J/g-MCL・min) was measured under various AMF irradiation conditions by changing output power (kW) and distance from irradiation surface (mm). Treatment condition for complete regression of animal tumors with 7 mm diameter was reproduced and heat dose in vivo (J/cm3) was calculated by multiplying heat generation activity (J/g-MCL・min) with MCL dosage (g-MCL/cm3) and irradiation time (min). Heat dose for tumor regression was revealed around 700-850 J/cm3 in every thrice AMF irradiations of a course. Since temperature-based treatments of large tumors were reported to fall into insufficiency, revealed heat dose was applied to design treatment condition of large tumor with 13-16 mm diameter, and complete regression was achieved by a course treatment. MCL dosage of temperature-based condition was found far lower than that of heat dose. Low MCL dosages (g-MCL/cm3) would cause shortage of heat dose (J/cm3) and insufficient anticancer activity, although tumor temperature could be raised by heat transfer to monitoring sites. These results showed utility of heat dose in vivo as index to ensure clinical efficacy and concomitantly to make useless invasive probe for temperature monitoring. Procedure to design treatment condition and required performance of AMF irradiator were described.

DOI： 10.3191/thermalmed.36.47

CiNii Research

若手化学工学者としての私の履歴書 Invited Reviewed

井藤　彰

学術の動向 Vol. 25 ( 7 ) page： 98 - 102 2020.7

A Curriculum Vitae of Mine as a Chemical Engineer

ITO Akira

TRENDS IN THE SCIENCES Vol. 25 ( 7 ) page： 7_98 - 7_102 2020.7

Heat dose index in vivo for cancer therapy using heat-generating nanoparticles named magnetite cationic liposomes and alternating magnetic field irradiator Reviewed

Tomio Morino, Shota Tanoue, Shuichiro Miyata, Kotaro Hirayama, Akira Ito, Toshiki Etani, Taku Naiki, Noriyasu Kawai, Takahiro Yasui

Vol. 36 ( 2 ) page： 47 - 58 2020.7

Novel neuromuscular junction model in 2D and 3D myotubes co-cultured with induced pluripotent stem cell-derived motor neurons. Reviewed

Kantaro Yoshioka, Akira Ito, Yoshinori Kawabe, Masamichi Kamihira

Journal of bioscience and bioengineering Vol. 129 ( 4 ) page： 486 - 493 2020.4

Language：English Publishing type：Research paper (scientific journal)

Motor neurons differentiated from induced pluripotent stem (iPS) cells have attracted attention for use in the construction of drug screening systems for neuronal diseases, such as amyotrophic lateral sclerosis. However, conventional drug screening systems using 2-dimensional (2D) cultures of iPS cell-derived motor neurons often evaluate the cell survival rate, morphological changes in the cells and/or gene expression analysis, and these parameters do not always reflect the actual functions of motor neurons, i.e., the induction of muscle contractions. In the present study, we developed a neuromuscular junction model comprising motor neurons and myotubes, which were differentiated from iPS cells and C2C12 myoblasts, respectively. Using this model, the contractile activity and force generation of the myotubes via the neuromuscular junction were successfully measured in both two- and three-dimensional (3D) cell culture systems. The results suggested that this neuromuscular junction model can be used to construct a drug candidate screening system for neuronal diseases.

DOI： 10.1016/j.jbiosc.2019.10.004

Targeted knock-in into the OVA locus of chicken cells using CRISPR/Cas9 system with homology-independent targeted integration. Reviewed

Ming Shi, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira

Journal of bioscience and bioengineering Vol. 129 ( 3 ) page： 363 - 370 2020.3

Language：English Publishing type：Research paper (scientific journal)

It is anticipated that transgenic avian species will be used as living bioreactors for the production of biopharmaceutical proteins. Precise tissue-specific expression of exogenous genes is a major challenge for the development of avian bioreactors. No robust vector is currently available for highly efficient and specific expression. In recent years, genome-editing techniques such as the CRISPR/Cas9 system have emerged as efficient and user-friendly genetic modification tools. Here, to apply the CRISPR/Cas9 system for the development of transgenic chickens, guide RNA sequences (gRNAs) of the CRISPR/Cas9 system for the ovalbumin (OVA) locus were evaluated for the oviduct-specific expression of exogenous genes. An EGFP gene expression cassette was introduced into the OVA locus of chicken DF-1 and embryonic fibroblasts using the CRISPR/Cas9 system mediated by homology-independent targeted integration. For the knock-in cells, EGFP expression was successfully induced by activation of the endogenous OVA promoter using the dCas9-VPR transactivation system. The combination of gRNAs designed around the OVA TATA box was important to induce endogenous OVA gene expression with high efficiency. These methods provide a useful tool for studies on the creation of transgenic chicken bioreactors and the activation of tissue-specific promoters.

DOI： 10.1016/j.jbiosc.2019.09.011

Medium temperature independent cytotoxicity of cell-adhesive heat-generating nanoparticles named magnetite cationic liposomes and its therapeutic use Reviewed

Tomio Morino, Hiroshi Takase, Toshiki Etani, Taku Naiki, Noriyasu Kawai, Akira Ito, Takahiro Yasui

Thermal Medicine Vol. 36 ( 1 ) page： 25 - 34 2020.3

Cre-Mediated Transgene Integration in Chinese Hamster Ovary Cells Using Minicircle DNA Vectors. Reviewed International journal

Wang X, Kawabe Y, Hada T, Ito A, Kamihira M

Biotechnology journal Vol. 13 ( 7 ) page： e1800063 2018.7

Language：English Publishing type：Research paper (scientific journal)

Bacterial backbone sequences of conventional plasmid vectors have been reported to exhibit negative effects on transgene expression in mammalian cells, such as cytotoxicity and gene silencing. Minicircle DNA vectors can be employed to overcome these issues and to improve the transfection efficiency because of their smaller size. In this study, transgenes are integrated into the hypoxanthine phosphoribosyltransferase (hprt) locus of Chinese hamster ovary (CHO) cells by the Cre-loxP system using minicircle DNA vectors as transgene donors. The targeted transgene integration efficiency is improved 2-3-fold (≈1.4%) using minicircle DNA vectors compared with conventional plasmid vectors. Moreover, clones with expected structures after transgene integration are obtained with a high frequency. When a transgene together with bacterial sequences derived from a plasmid vector is integrated into the hprt locus, the cell growth rate and antibody titer decrease. These results indicate that minicircle DNA vectors are more suitable than conventional plasmid vectors for transgene delivery in recombinant protein production using CHO cells.

DOI： 10.1002/biot.201800063

Targeted knock-in of an scFv-Fc antibody gene into the hprt locus of Chinese hamster ovary cells using CRISPR/Cas9 and CRIS-PITCh systems Reviewed

Yoshinori Kawabe, Shinya Komatsu, Shodai Komatsu, Mai Murakami, Akira Ito, Tetsushi Sakuma, Takahiro Nakamura, Takashi Yamamoto, Masamichi Kamihira

Journal of Bioscience and Bioengineering Vol. 125 ( 5 ) page： 599 - 605 2018.5

Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier B.V.

Chinese hamster ovary (CHO) cells have been used as host cells for the production of pharmaceutical proteins. For the high and stable production of target proteins, the transgene should be integrated into a suitable genomic locus of host cells. Here, we generated knock-in CHO cells, in which transgene cassettes without a vector backbone sequence were integrated into the hprt locus of the CHO genome using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 and CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) systems. We investigated the efficiency of targeted knock-in of transgenes using these systems. As a practical example, we generated knock-in CHO cells producing an scFv-Fc antibody using the CRIS-PITCh system mediated by microhomology sequences for targeting. We found that the CRIS-PITCh system can facilitate targeted knock-in for CHO cell engineering.

DOI： 10.1016/j.jbiosc.2017.12.003

Characterization of genetically engineered mouse hepatoma cells with inducible liver functions by overexpression of liver-enriched transcription factors Reviewed

Hideaki Yamamoto, Jane Marie Tonello, Takanori Sambuichi, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira

Journal of Bioscience and Bioengineering Vol. 125 ( 1 ) page： 131 - 139 2018.1

Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier B.V.

New cell sources for the research and therapy of organ failure could significantly alleviate the shortage of donor livers that are available to patients who suffer from liver disease. Liver carcinoma derived cells, or hepatoma cells, are the ideal cells for developing bioartificial liver systems. Such cancerous liver cells are easy to prepare in large quantities and can be maintained over long periods under standard culture conditions, unlike primary hepatocytes. However, hepatoma cells possess only a fraction of the functions of primary hepatocytes. In a previous study, by transducing cells with liver-enriched transcription factors that could be inducibly overexpressed—hepatocyte nuclear factor (HNF)1α, HNF1β, HNF3β [FOXA2], HNF4α, HNF6, CCAAT/enhancer binding protein (C/EBP)α, C/EBPβ and C/EBPγ—we created mouse hepatoma cells with high liver-specific gene expression called the Hepa/8F5 cell line. In the present study, we performed functional and genetic analyses to characterize the Hepa/8F5 cell line. Further, in three-dimensional cultures, the function of these cells improved significantly compared to parental cells. Ultimately, these cells might become a new resource that can be used in basic and applied hepatic research.

DOI： 10.1016/j.jbiosc.2017.07.011

温熱誘導型遺伝子発現システムの開発と応用 Invited Reviewed

井藤　彰

放射線生物研究 Vol. 53 ( 2 ) page： 104-114 2018

ハイパーサーミアに伴う免疫賦活のメカニズム Invited Reviewed

井藤彰

放射線生物研究 Vol. 53 ( 2 ) page： 104-114 2018

Three-dimensional culture of a genetically modified hepatoma cell line using macroporous gelatin beads Reviewed

Jane Marie Tonello, Saori Kawashima, Kazuki Sato, Yoshinori Kawabe, Akira Ito, Masamichi Kamihira

CYTOTECHNOLOGY Vol. 69 ( 6 ) page： 925 - 931 2017.12

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER

Hepatoma cells are a candidate cell source for bio-artificial livers. However, they exhibit reduced liver functions compared with primary hepatocytes. In our previous study, genetically engineered mouse hepatoma cells were created by transduction with vectors mediating inducible overexpression of eight liver-enriched transcription factors. Upon the induction of the liver-enriched transcription factors transduced, the cells expressed both phenotypic and genotypic liver functions at high levels. In the present study, we performed three-dimensional culture of these cells using macroporous gelatin beads. When immobilized on the macroporous gelatin beads, these cells exhibited further enhancement in liver functionality, including increased albumin secretion, ammonia removal and cytochrome P450 activity. The levels of these functions were significantly enhanced compared to monolayer culture. The method is simple and scalable, and provides highly functional cells that can be used in basic and applied fields of hepatic research.

DOI： 10.1007/s10616-017-0117-0

Accumulative scFv-Fc antibody gene integration into the &IThprt&IT chromosomal locus of Chinese hamster ovary cells Reviewed

Xue Wang, Yoshinori Kawabe, Risa Kato, Takeshi Hada, Akira Ito, Yoshimasa Yamana, Masako Kondo, Masamichi Kamihira

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 124 ( 5 ) page： 583 - 590 2017.11

Language：English Publishing type：Research paper (scientific journal) Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN

We have previously developed an accumulative site-specific gene integration system (AGIS) using Cre-recombinase and mutated loxP sites. AGIS enables repeated transgene integration into a predetermined chromosomal site in mammalian cells. However, the process of establishing cells with multiple integrated copies of the transgene is still time-consuming. In the present study, we describe an improved version of AGIS that facilitates and accelerates the establishment of high-producer Chinese hamster ovary (CHO) cells. Two donor vectors were simultaneously introduced into the cells in a single transfection. Cells with successfully targeted transgene integration were screened based on a change in the color of the reporter fluorescent protein that they express. Repeated rounds of integration allowed the transgene copy number to be increased. As a model, an scFv-Fc antibody gene was integrated into the hprt locus of the CHO cell genome. After three rounds of integration, a high-producer CHO cell clone with six copies of the scFv-Fc gene was successfully established. scFv-Fc productivity was approximately four-fold greater than a control cell line harboring a single copy of the transgene. This newly designed AGIS procedure should facilitate the development of producer cells suitable for biopharmaceutical protein production. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2017.05.017

Hypoxia-responsive transgene expression system using RTP801 promoter and synthetic transactivator fused with oxygen-dependent degradation domain Reviewed

Akihiko Ono, Akira Ito, Tomonaga Sato, Masaki Yamaguchi, Taiga Suzuki, Yoshinori Kawabe, Masamichi Kamihira

JOURNAL OF BIOSCIENCE AND BIOENGINEERING Vol. 124 ( 1 ) page： 115 - 124 2017.7

Language：English Publishing type：Research paper (scientific journal) Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN

Precise control of gene expression using an artificial gene circuit is a major challenge in the application of synthetic biology. Here, we designed a hypoxia-responsive transgene expression system by combining a hypoxia-inducible RTP801 promoter and a tetracycline-responsive transactivator fused with an oxygen-dependent degradation domain (TA-ODD). The reporter gene expression was highly induced by hypoxia when a transactivator-expression plasmid, pRTP801/TA-ODD, harboring a TA-ODD gene driven by the RTP801 promoter, was cotransfected with a reporter plasmid, pTRE/EGFP, harboring an EGFP gene controlled under the transactivator-responsive promoter. A stable cell line into which the expression units RTP801/TA-ODD and THE/EGFP had been introduced responded to hypoxia and expressed the reporter gene in an oxygen-concentration-dependent manner. Moreover, the cells demonstrated potential as sensors to detect hypoxic conditions in a three-dimensional tissue culture in vitro. These results indicate that the hypoxia-responsive transgene expression system is useful for constructing cell-based hypoxia detection systems. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2017.02.012

Novel therapy for pancreatic fistula using adipose-derived stem cell sheets treated with mannose Reviewed

Kaneko Hirokazu, Kokuryo Toshio, Yokoyama Yukihiro, Yamaguchi Junpei, Yamamoto Tokunori, Shibata Rei, Gotoh Momokazu, Murohara Toyoaki, Ito Akira

SURGERY Vol. 161 ( 6 ) page： 1561 - 1569 2017.6

Effects of heat stimulation and L-ascorbic acid 2-phosphate supplementation on myogenic differentiation of artificial skeletal muscle tissue constructs Reviewed

Kazushi Ikeda, Akira Ito, Masanori Sato, Shota Kanno, Yoshinori Kawabe, Masamichi Kamihira

JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE Vol. 11 ( 5 ) page： 1322 - 1331 2017.5

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY

Although skeletal muscle tissue engineering has been extensively studied, the physical forces produced by tissue-engineered skeletal muscles remain to be improved for potential clinical utility. In this study, we examined the effects of mild heat stimulation and supplementation of a L-ascorbic acid derivative, L-ascorbic acid 2-phosphate (AscP), on myoblast differentiation and physical force generation of tissue-engineered skeletal muscles. Compared with control cultures at 37 degrees C, mouse C2C12 myoblast cells cultured at 39 degrees C enhanced myotube diameter (skeletal muscle hypertrophy), whereas mild heat stimulation did not promote myotube formation (differentiation rate). Conversely, AscP supplementation resulted in an increased differentiation rate but did not induce skeletal muscle hypertrophy. Following combined treatment with mild heat stimulation and AscP supplementation, both skeletal muscle hypertrophy and differentiation rate were enhanced. Moreover, the active tension produced by the tissue-engineered skeletal muscles was improved following combined treatment. These findings indicate that tissue culture using mild heat stimulation and AscP supplementation is a promising approach to enhance the function of tissue-engineered skeletal muscles. Copyright (C) 2015 John Wiley & Sons, Ltd.

DOI： 10.1002/term.2030