Cell Medicine 5(2-3) Abstracts

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Cell Medicine, Vol. 5, pp. 45–51, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666549
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Bioimaging of Transgenic Rats Established at Jichi Medical University: Applications in Transplantation Research

Takumi Teratani and Eiji Kobayashi

Division of Development of Advanced Therapy, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsukeshi, Tochigi, Japan

Research in the life sciences has been greatly advanced by the ability to directly visualize cells, tissues, and organs. Preclinical studies often involve many small and large animal experiments and, frequently, cell and organ transplantations. The rat is an excellent animal model for the development of transplantation and surgical techniques because of its small size and ability to breed in small spaces. Ten years ago, we established colorimaging transgenic rats and methods for the direct visualization of their tissues. Since then, our transgenic rats have been used throughout the various fields that are concerned with cell transplantation therapy. In this minireview, we summarize results from some of the groups that have used our transgenic rats at the bench level and in cell transplantation research.

Key words: Cell transplantation; Transgenic rat; Bioimaging; Human injury models

Received June 1, 2012; final acceptance August 20, 2013. Online prepub date: August 29, 2013.
Address correspondence to Eiji Kobayashi, M.D., Ph.D., Division of Development of Advanced Therapy, Center for Development of Advanced Medical Technology, Jichi Medical University, 3311-1 Yakushiji, Shimotsukeshi, Tochigi 329-0498, Japan. Tel: +81-285-58-7583; Fax: +81-285-44-6219; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 53–57, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666512
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

ER Stress and β-Cell Pathogenesis of Type 1 and Type 2 Diabetes and Islet Transplantation

Hitomi Usui Kataoka* and Hirofumi Noguchi†

*Department of Primary Care and Medical Education, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
†Department of Surgery, Clinical Research Center, Chiba-East Hospital, National Hospital Organization, Chiba, Japan

Endoplasmic reticulum (ER) stress affects the pathogenesis of diabetes. ER stress plays important roles, both in type 1 and type 2 diabetes, because pancreatic β-cells possess highly developed ER for insulin secretion. This review summarizes the relationship between ER stress and the pathogenesis of type 1 and type 2 diabetes. In addition, the association between islet transplantation and ER stress is discussed.

Key words: β-Cells; Endoplasmic reticulum (ER) stress; Islet transplantation

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Hitomi Usui Kataoka, M.D., Ph.D., Department of Primary Care and Medical Education, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. Tel: +81-86-235-6963; Fax: +81-86-221-6963; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Hirofumi Noguchi, M.D., Ph.D., Department of Surgery, Clinical Research Center, Chiba-East National Hospital, National Hospital Organization, 673 Nitona, Chuo-ku, Chiba 260-8712, Japan. Tel: +81-43-261-5171; Fax: +81-43-268-2613; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 59–62, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666558
Copyright © 2013 Cognizant Comm. Corp.
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Review

A Review of Autologous Islet Transplantation

Michihiro Maruyama, Takashi Kenmochi, Naotake Akutsu, Kazunori Otsuki, Taihei Ito, Ikuko Matsumoto, and Takehide Asano

Department of Surgery, NHO Chiba-East National Hospital, Chiba City, Chiba, Japan

Autologous islet transplantation after total or semitotal pancreatectomy aims to preserve insulin secretory function and prevent the onset of diabetes. The major indication for pancreatectomy is chronic pancreatitis with severe abdominal pain, a benign pancreatic tumor, and trauma. The metabolic outcome of autologous islet transplantation is better than that of allogeneic transplantation and depends on the number of transplanted islets. Achieving islet isolation from a fibrous or damaged pancreas is one of the biggest challenges of autologous islet transplantation; a major complication is portal vein thrombosis after crude islet infusion. However, the incidence of portal vein thrombosis has decreased as islet preparation techniques have improved over time.

Key words: Total pancreatectomy; Islet; Autologous transplantation

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Michihiro Maruyama, Department of Surgery, NHO Chiba-East National Hospital, 673 Nitona, Chuo-ku, Chiba City, Chiba 2608712, Japan. Tel: +81-43-261-5171; Fax: +81-43-268-2613; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 63–68, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666495
Copyright © 2013 Cognizant Comm. Corp.
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Culture Conditions for Mouse Pancreatic Stem Cells

Hirofumi Noguchi,* Issei Saitoh,† Hitomi Usui Kataoka,‡ Masami Watanabe,§ Yasufumi Noguchi,¶ and Toshiyoshi Fujiwara#

*Department of Surgery, Clinical Research Center, Chiba-East Hospital, National Hospital Organization, Chiba, Japan
†Department of Pediatric Dentistry, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan
‡Department of Primary Care and Medical Education, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
§Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
¶Department of Socio-environmental Design, Hiroshima International University, Hiroshima, Japan
#Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

Recently, mouse pancreatic stem cells have been isolated from adult mouse pancreata. However, these pancreatic stem cells could be maintained only under specific culture conditions with lot-limited fetal bovine serum (FBS). For the efficient isolation and maintenance of mouse pancreatic stem cells, it is important to identify culture conditions that can be used independent of the FBS lot. In this study, we evaluated the culture conditions required to maintain mouse pancreatic stem cells. The mouse pancreatic stem cells derived from the pancreas of a newborn mouse, HN#101, were cultured under the following conditions: 1) Dulbecco’s modified Eagle’s medium (DMEM) with 20% lot-limited FBS, in which mouse pancreatic stem cells could be cultured without changes in morphology and growth activity; 2) complete embryonic stem (ES) cell media; and 3) complete ES cell media on feeder layers of mitomycin C-treated STO cells, which were the same culture conditions used for mouse ES cells. Under culture conditions #1 and #3, the HN#101 cells continued to form a flat “cobblestone” monolayer and continued to divide actively beyond the population doubling level (PDL) 100 without growth inhibition, but this did not occur under culture condition #2. The gene expression profile and differentiated capacity of the HN#101 cells cultured for 2 months under culture condition #3 were similar to those of HN#101 cells at PDL 50. These data suggest that complete ES cell media on feeder layers could be useful for maintaining the undifferentiated state of pancreatic stem cells.

Key words: Mouse pancreatic stem cells; Culture condition; Embryonic stem cells; Embryonic stem (ES) medium; Feeder cells; Pancreatic islet transplantation

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Hirofumi Noguchi, M.D., Ph.D., Department of Surgery, Clinical Research Center, Chiba-East National Hospital, National Hospital Organization, 673 Nitona, Chuo-ku, Chiba 260-8712, Japan. Tel: +81-43-261-5171; Fax: +81-43-268-2613; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 69–73, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666503
Copyright © 2013 Cognizant Comm. Corp.
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Isolation Efficiency of Mouse Pancreatic Stem Cells Is Age Dependent

Takashi Kuise,* Hirofumi Noguchi,† Issei Saitoh,‡ Hitomi Usui Kataoka,§ Masami Watanabe,¶ Yasufumi Noguchi,# and Toshiyoshi Fujiwara*

*Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
†Department of Surgery, Clinical Research Center, Chiba-East Hospital, National Hospital Organization, Chiba, Japan
‡Department of Pediatric Dentistry, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan
§Department of Primary Care and Medical Education, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
¶Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
#Department of Socio-environmental Design, Hiroshima International University, Hiroshima, Japan

Mouse pancreatic stem cells have been isolated from mouse pancreata. This study evaluated the efficacy of isolating mouse pancreatic stem cells using mice of different ages. The pancreata of newborn mice, 8-week-old mice, and 24-week-old mice were harvested and digested by using collagenase. The “duct-like” cells in the digested pancreatic tissue were then inoculated into 96-well plates, cloned by limiting dilution, and cultured in DMEM with 20% FBS. Pancreatic stem cells were isolated from the pancreata of all newborn mice, while cells could only be isolated from 10% of the pancreata of 8-week-old mice and could not be isolated from the pancreata of any 24-week-old mice. These data suggest that young mice may have some pancreatic stem cells and that older mice may only have a few pancreatic stem cells. These data also indicate that it is extremely difficult to isolate pancreatic stem cells from older mice, suggesting that future research focus its efforts on finding methods of isolating pancreatic stem cells from adult mice.

Key words: Mouse pancreatic stem cells; Age dependent; Diabetes; ES medium; Feeder cells; Pancreatic islet transplantation

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Hirofumi Noguchi, M.D., Ph.D., Department of Surgery, Clinical Research Center, Chiba-East National Hospital, National Hospital Organization, 673 Nitona, Chuo-ku, Chiba 260-8712, Japan. Tel: +81-43-261-5171; Fax: +81-43-268-2613; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 75–81, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666477
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Use of Mesenchymal Stem Cell-Conditioned Medium to Activate Islets in Preservation Solution

Naoya Kasahara,*†1 Takumi Teratani,*1 Junshi Doi,* Yuki Iijima,* Masashi Maeda,* Shinji Uemoto,‡ Yasuhiro Fujimoto,*‡ Naohiro Sata,† Yoshikazu Yasuda,† and Eiji Kobayashi*

*Division of Development of Advanced Therapy, Center for Development of Advanced Medical Technology, Jichi Medical University, Shimotsukeshi, Tochigi, Japan
†Department of Surgery, Jichi Medical University, Shimotsukeshi, Tochigi, Japan
‡Division of Hepato-Pancreato-Biliary Surgery and Transplantation, Department of Surgery, Kyoto University Graduate School of Medicine, Syougoin, Sakyoku, Kyotoshi, Kyoto, Japan

Pancreatic islet transplantation has received widespread attention as a promising treatment for type 1 diabetes. However, islets for transplantation are subject to damage from a number of sources, including ischemic injury during removal and delivery of the donor pancreas, enzymatic digestion during islet isolation, and reperfusion injury after transplantation in the recipient. Here we found that protein fractions secreted by mesenchymal stem cells (MSCs) were capable of activating preserved islets. A conditioned medium from the supernatant obtained by culturing adipose tissue MSCs (derived from wild-type Lewis rats) was prepared for 2 days in serum-free medium. Luc-Tg rat islets to which an organ preservation solution was added were then incubated at 4°C with fractions of various molecular weights prepared from the conditioned medium. Under the treatment with some of the fractions, by 4 days the relative luminescence intensities (representative of the ATP levels of the cold-preserved islets) had increased to over 150% of their initial values. Our novel system may be able to restore isolated islets to the condition they were in before transport, culture, and transplantation.

Key words: Preservation of islets; Mesenchymal stem cells (MSCs); Transgenic rat; Living image

Received June 1, 2013; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Takumi Teratani, Ph.D., Division of Development Advanced Therapy, Center for Development of Advanced Medical Technology, Jichi Medical University, 3311-1 Yakushiji, Shimotsukeshi, Tochigi 329-0498, Japan. Tel.: +81-285-58-7583; Fax: +81-285-44-6219; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 83–87, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666486
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Inhibition of Hepatic Ischemic Reperfusion Injury Using Saline Exposed to Electron Discharge in a Rat Model

Masaharu Dozen,*† Shin Enosawa,* Yuki Tada,† and Keisuke Hirasawa†
*Division for Advanced Medical Sciences, Clinical Research Center, National Center for Child Health and Development, Setagaya-ku, Tokyo, Japan
†Research Unit, Cambwick Healthcare Corporation, Akishima City, Tokyo, Japan

The pathogenesis of ischemia/reperfusion (I/R) injury in surgical trauma, organ transplantations, and brain and myocardial infarctions is attributable to excessive oxidative stress caused by reactive oxygen species and their metabolites. We prepared a physiological saline solution treated with simple exposure to a microampere current with electron discharge onto the surface; this treatment increased reduction potential and caused proton reduction. We examined the reductive potency of the electron-treated solution (ETS) on hepatocellular I/R injury in a rat model. When the ETS was administered in four doses at 0, 3, 10, and 20 min after reperfusion, severe hepatocellular injury was suppressed, as revealed by a decrease in serum alanine aminotransferase levels and histopathological improvement of liver damage. Since a preparation of hydrogen gas-dissolved saline solution (HDS) was shown to be capable of suppressing I/R injury, the possible involvement of dissolved hydrogen gas in the effectiveness of ETS was examined. When HDS was treated by degasification, the aforementioned effectiveness was decreased. In contrast, the same treatment did not alter the effectiveness of ETS. These results suggest that the antioxidative efficacy of ETS is not attributable to dissolved hydrogen gas but presumably to the molecule(s) produced from the stepwise reduction of protons in the solution.

Key words: Antioxidation; Electron; Ischemia; Reperfusion; Tissue injury

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Keisuke Hirasawa, Research Unit, Cambwick Healthcare Corporation, 3-6-1, Azuma-cho, Akishima City, Tokyo 196-0033, Japan. Tel: +81-42-519-1171; Fax: +81-42-519-1172; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 89–96, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666530
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Observation of Positively Charged Magnetic Nanoparticles Inside HepG2 Spheroids Using Electron Microscopy

Yoshitaka Miyamoto,* Yumie Koshidaka,* Hirofumi Noguchi,† Koichi Oishi,* Hiroaki Saito,‡ Hiroshi Yukawa,* Noritada Kaji,§ Takeshi Ikeya,¶ Satoshi Suzuki,# Hisashi Iwata,** Yoshinobu Baba,§ Katsutoshi Murase,‡ and Shuji Hayashi*

*Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Japan
†Department of Surgery, Clinical Research Center, Chiba-East Hospital, National Hospital Organization, Chiba, Japan
‡Nagoya Research Laboratory, MEITO Sangyo Co., Ltd., Nishibiwajima, Kiyosu, Japan
§Department of Applied Chemistry, Nagoya University Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya, Japan
¶Photosensitive Materials Research Center, Toyo Gosei Co., Ltd., Kamimyoden, Ichikawa, Japan
#Research Laboratories, HAB Research Organization, Ichikawa General Hospital, Chiba, Japan
**Department of Orthopaedic Surgery, Nagoya Kyoritsu Hospital, Nakagawa-ku, Nagoya, Japan

Magnetic resonance imaging (MRI) using magnetic nanoparticles has been used to diagnose vascular diseases as well as to monitor transplanted cells and tissues. In this study, we synthesized magnetic iron oxide nanoparticles (TMADM-03), electrically charged by the presence of a cationic end-group substitution of dextran, and observed these nanoparticles inside three-dimensional models of HepG2 spheroids, which mimic tissues. Patterned cell array glass disks were prepared to visualize the presence of TMADM-03 uptaken by HepG2 spheroids using transmission electron microscopy (TEM). The HepG2 cells (2 × 105 cells) were inoculated onto Cell-able™ 12-well plates. After 48 h of culture, the cells were incubated with 75 μg Fe/ml TMADM-03 in culture medium for 24 h. To investigate the cellular function of the HepG2 spheroids, the albumin secretion was evaluated by an ELISA. The albumin secretion after incubation for 24 h was reduced compared with the secretion prior to the addition of TMADM-03. TEM image samples were prepared in a planar direction or a vertical direction to the HepG2 spheroids on patterned cell array glass disks. The incorporation of TMADM-03 inside the HepG2 spheroids was confirmed. In addition, TMADM-03 could be observed in the deeper layers of the spheroids, and this was localized in the lysosomes. These data suggest that the novel magnetic iron oxide nanoparticles invade three-dimensional HepG2 spheroids.

Key words: Magnetic iron oxide nanoparticle; Cationic dextran; Human hepatocellular carcinoma cells (HepG2); Spheroids; Electron microscopy

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Yoshitaka Miyamoto, Ph.D., Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. Tel: +81-52-719-1873; Fax: +81-52-719-1977; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 5, pp. 97–101, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X666521
Copyright © 2013 Cognizant Comm. Corp.
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Improved Recovery of Hepatocytes Isolated From Warm Ischemic Rat Liver by Citrate Phosphate Dextrose (CPD)-Supplemented Euro-Collins Solution

Huai-Che Hsu,*† Naoto Matsuno,* Noboru Machida,† and Shin Enosawa*

*Clinical Research Center, National Center for Child Health and Development, Tokyo, Japan
†Department of Veterinary Surgery, Faculty of Veterinary Medicine, Tokyo University of Agriculture and Technology, Tokyo, Japan

Demand for human primary hepatocytes is increasing, particularly for clinical trials of hepatocyte transplantation. However, due to the severe shortage of organ transplant donors, the source of cells for these endeavors is restricted to untransplantable livers, such as those from non-heart-beating donors and surgically resected liver tissues. To improve cell recovery from such sources after warm ischemia, we evaluated the efficacy of applying perfusion solutions, focusing on improvement of hepatocyte recovery. Warm ischemia was induced by clamping both portal vein and hepatic artery for 10 or 15 min in rats. The liver was perfused with either Euro-Collins (EC) or extracellular-type trehalose-containing Kyoto (ETK) solutions supplemented with an anticoagulant, either heparin or citrate phosphate dextrose solution (CPD), compared to Ca2+, Mg2+-free Hanks solution. While the viability of recovered cells was 81.5 ± 4.2% and cell yield was 2.27 ± 0.53 × 108 in nonwarm ischemia controls (n = 11), these values were only 74.7 ± 2.9% and 0.38 ± 0.17 × 108, respectively, in the 10-min warm ischemia group, using the Hanks as the perfusion solution. Although the addition of heparin increased the live cell number only twofold (0.71 ± 0.40 × 108, n = 4), the best improvement was achieved by adding CPD to EC. This resulted in a recovery of 1.41 ± 0.50 × 108 in the 10-min ischemia group (n = 7) and 1.37 ± 0.28 × 108 in the 15-min group (n = 3). Macroscopic observation showed that blood had been completely flushed out by the solution, suggesting good restoration of the microcirculation in ischemic liver. Using ETK instead of EC resulted in a slight decrease in efficacy. These results demonstrate that CPD, as opposed to heparin, is effective in ensuring liver microcirculation and flushing out the blood and that EC is the best perfusion solution for obtaining hepatocytes from ischemic liver.

Key words: Hepatocyte isolation; Warm ischemia; Citrate

Received June 1, 2012; final acceptance May 1, 2013. Online prepub date: May 14, 2013.
Address correspondence to Shin Enosawa, Division Chief, Division for Advanced Medical Sciences, Clinical Research Center, National Center for Child Health and Development, 2-10-1 Ookura, Setagaya-ku, Tokyo 157-8534, Japan. Tel: +81-3-5494-8163; Fax: +81-3-3417-2864; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it