Cell Medicine 6(1-2) Abstracts

Return to Cell Medicine page>

Cell Medicine, Vol. 6, pp. 3–8, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674171
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of New Preservation Solutions, HN-1 and University of Wisconsin Solution, in Pancreas Preservation for Porcine Islet Isolation

Akihiro Katayama,* Hirofumi Noguchi,† Takashi Kuise,‡ Atsuko Nakatsuka,* Daisho Hirota,* Hitomi Usui Kataoka,§ Takashi Kawai,‡ Kentaro Inoue,* Noriko Imagawa,‡ Issei Saitoh,¶ Yasufumi Noguchi,# Masami Watanabe,** Jun Wada,* and Toshiyoshi Fujiwara‡

*Department of Medicine and Clinical Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan
†Department of Surgery, Chiba-East National Hospital, National Hospital Organization, Chiba 260-8712, Japan
‡Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
§Department of Primary Care and Medical Education, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
¶Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University, Niigata, Japan
#Department of Socio-environmental Design, Hiroshima International University, Hiroshima, Japan
**Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

For pancreatic islet transplantation, maintaining organ viability after pancreas procurement is critical and a major determinant for better graft function and survival. University of Wisconsin (UW) solution is currently the gold standard for abdominal organ preservation and the pancreas in particular. However, in the use of UW preservation solution for islet transplantation, there are disadvantages to be overcome, such as the inhibition of collagenase activity during pancreatic digestion. In this study, we compared UW solution with HN-1 solution in pancreas preservation for islet isolation. Islet yield was significantly greater in the HN-1 group than the UW group both before and after purification. In the in vitro assay, the adenosine triphosphate content in cultured islets was significantly higher in the HN-1 group than in the UW group. Furthermore, in streptozotocin-induced diabetic nude mice, the islet graft function of the HN-1 group was superior to that of the UW group. We concluded that the use of HN-1 solution is a promising approach for optimal pancreas preservation in islet transplantation.

Key words: Islet transplantation; Islet isolation; HN-1 solution; University of Wisconsin (UW) solution; Preservation

Received April 30, 2013; final acceptance October 2, 2013. Online prepub date: October 21, 2013.
Address correspondence to Hirofumi Noguchi, M.D., Ph.D., Department of Surgery, Chiba-East National Hospital, National Hospital Organization, 673 Nitona-cho, 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. 6, pp. 9–14, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674180
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of Incubation Solutions Prior to the Purification of Porcine Islet Cells

Takashi Kawai,* Hirofumi Noguchi,† Takashi Kuise,* Atsuko Nakatsuka,‡ Akihiro Katayama,‡ Noriko Imagawa,* Hitomi Usui Kataoka,§ Issei Saitoh,¶ Yasufumi Noguchi,# Masami Watanabe,** and Toshiyoshi Fujiwara*

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

For pancreatic islet transplantation, one of the most important steps of islet isolation is islet purification. The most common method of islet purification is density gradient centrifugation because there are differences in density between islets and acinar tissue. However, the density of islets/acinar tissue depends on several conditions, such as the incubation time before purification and the osmolality of the preincubation solution. In this study, we evaluated the impact of using two different preincubation solutions before purification. We used the University of Wisconsin (UW) solution and a new preservation solution (HN-1), which we recently developed. There were no significant differences between the two solutions in terms of the islet yield, rate of viability, and purity or stimulation index after purification. There were also no differences in the attainability and suitability of posttransplantation normoglycemia. Our study shows that the HN-1 solution is equivalent to the UW solution for preincubation before islet purification.

Key words: Islet transplantation; Islet isolation; University of Wisconsin (UW) solution; HN-1 solution; Preincubation

Received April 30, 2013; final acceptance October 2, 2013. Online prepub date: October 21, 2013.
Address correspondence to Hirofumi Noguchi, M.D., Ph.D., Department of Surgery, Chiba-East National Hospital, National Hospital Organization, 673 Nitona-cho, 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. 6, pp. 15–23, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674199
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Maintenance of Viability and Function of Rat Islets With the Use of ROCK Inhibitor Y-27632

Yasuhiro Kubota,* Hirofumi Noguchi,† Masayuki Seita,* Takeshi Yuasa,* Hiromi Sasamoto,* Shuhei Nakaji,‡ Teru Okitsu,§ Toshiyoshi Fujiwara,* and Naoya Kobayashi*

*Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
†Department of Surgery, Chiba-East National Hospital, National Hospital Organization, Chiba, Japan
‡Department of Biomedical Engineering, Okayama University of Science, Okayama, Japan
§Department of Organ Transplantation Center, Kyoto University, Kyoto, Japan

The number of patients with diabetes is on an increasing trend, thus leading to the belief that diabetes will be the largest medical problem of the 21st century. Islet transplantation can improve glycometabolic control in patients with type 1 diabetes. We studied the viability of Rho-associated protein kinase (ROCK) inhibitor Y-27632 in a culture system in vitro on freshly isolated rat islets. Islet isolation was conducted on a Lewis rat, and studies of culture solutions were split into two groups, one group using ROCK inhibitor Y-27632, and another without. On the seventh day of culture, we evaluated the differences for the cell morphology, viability, and insulin secretion. The Y-27632 group maintained form better than the group without Y-27632. With strong expression of Bcl-2 observed with the Y-27632 group, and expression suppressed with Bax, inhibition of apoptosis by Y-27632 was confirmed. The Y-27632 group predominantly secreted insulin. For islet transplantation, Y-27632 inhibited cell apoptosis in a graft and was also effective in promoting insulin secretion. We were able to confirm effective morphological and functional culture maintenance by separating islets from a rat and adding ROCK inhibitor Y-27632 to the medium.

Key words: Islet transplantation; Apoptosis; Embryonic stem cells; Induced pluripotent stem cells; Rho-associated protein kinase (ROCK) inhibitor; Y-27632

Received April 30, 2013; final acceptance October 1, 2013. Online prepub date: October 21, 2013.
Address correspondence to Hirofumi Noguchi, M.D., Ph.D., Department of Surgery, Chiba-East National Hospital, National Hospital Organization, 673 Nitona-cho, 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. 6, pp. 25–31, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674289
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Development of Canine Models of Type 1 Diabetes With Partial Pancreatectomy and the Administration of Streptozotocin

Masayuki Seita,* Hirofumi Noguchi,† Yasuhiro Kubota,* Hironobu Kawamoto,* Shuhei Nakaji,‡ Naoya Kobayashi,* 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 National Hospital, National Hospital Organization, Chiba, Japan
‡Department of Biomedical Engineering, School of Engineering, Okayama University of Science, Okayama, Japan

We created canine models of type 1 diabetes that were suitable for the assessment of cell therapies, such as islet transplantation and bioartificial pancreas, with low-dose streptozotocin (STZ) injection and partial pancreatectomy. In our model, a 50% pancreatectomy was performed with general anesthesia, followed by systemic injection of 35 mg/kg STZ into a vein of the foreleg. Four weeks after the administration of STZ, the fasting blood glucose level of our model dogs was found to be over 200 mg/dl twice on different days, and we could not detect any canine insulin by the intravenous glucose tolerance test (IVGTT). We therefore diagnosed the dogs to have induced diabetes. Some studies have reported high-dose STZ to be very toxic for both the kidney and liver, and therefore a lower dose is desirable to induce diabetic models without any associated kidney or liver damage. We think that the combination of a partial pancreatectomy can thus make it possible to reduce the dose of STZ, and it is therefore useful for the creation of type 1 diabetes models. We believe that our model is a safe and reliable model for type 1 diabetes in canines to assess the efficacy of pancreas-targeted cell therapies.

Key words: Pancreatectomy; Streptozotocin (STZ); Diabetes; Dogs

Received April 30, 2013; final acceptance October 1, 2013. Online prepub date: October 21, 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-cho, 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. 6, pp. 33–38, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674243
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Quality of Air-Transported Human Islets for Single Islet Cell Preparations

Shingo Yamashita,* Kazuo Ohashi,*† Rie Utoh,* Tatsuya Kin,‡ A. M. James Shapiro,‡ Masakazu Yamamoto,† Mitsukazu Gotoh,§ and Teruo Okano*

*Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, Tokyo, Japan
†Department of Surgery, Institute of Gastroenterology, Tokyo Women’s Medical University, Tokyo, Japan
‡Clinical Islet Transplant Program, University of Alberta, Edmonton, Alberta, Canada
§Department of Regenerative Surgery, Fukushima Medical University, Fukushima, Japan

In new generation medical therapies for type 1 diabetes mellitus (DM), cell-based approaches using pancreatic islets have attracted significant attention worldwide. In particular, dispersed islet cells obtained from isolated pancreatic islets have been a valuable source in the cell biology and tissue engineering fields. Our experimental approach to the development of new islet-based DM therapies consisted of creating a monolithic islet cell sheet format using dispersed islet cells. In this experiment, we explored the potential of internationally transporting human islets from Alberta, Canada to Tokyo, Japan and obtaining viable dispersed islet cells. A total of 34 batches of isolated and purified human islets were transported using a commercial air courier service. Prior to shipping, the human islets had been in culture for 0–108 h at the University of Alberta. The transportation period from Alberta to Tokyo was 2–5 days. The transported human islet cells were enzymatically dispersed as single cells in Tokyo. The number of single islet cells decreased as the number of transportation days increased. In contrast, cell viability was maintained regardless of the number of transportation days. The preshipment culture time had no effect on the number or viability of single cells dispersed in Tokyo. When dispersed single islet cells were plated on laminin-5-coated temperature-responsive polymer-grafted culture dishes, the cells showed favorable attachment followed by extension as a monolithic format. The present study demonstrated that long-distance transported human islets are a viable cell source for experiments utilizing dispersed human islet cells.

Key words: Islet transportation; Dispersed islet cells; Temperature-responsive polymer-grafted culture dish; Laminin-5

Received April 30, 2013; final acceptance October 8, 2013. Online prepub date: October 23, 2013.
Address correspondence to Teruo Okano, Ph.D., Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. Tel: +81-3-5367-9945 ext. 6201; Fax: +81-3-3359-6046; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 6, pp. 39–45, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674207
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of the Pharmacological Efficacies of Immunosuppressive Drugs Evaluated by the ATP Production and Mitochondrial Activity in Human Lymphocytes

Hiroyasu Sasahara,* Kentaro Sugiyama,† Mahoto Tsukaguchi,* Kazuya Isogai,* Akira Toyama,* Hiroshi Satoh,* Kazuhide Saitoh,‡ Yuki Nakagawa,‡ Kota Takahashi,‡ Sachiko Tanaka,† Kenji Onda,† and Toshihiko Hirano†

*Division of Pharmacy, Niigata University Medical and Dental Hospital, Niigata, Japan
†Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Japan
‡Division of Urology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan

The lymphocyte immunosuppressant sensitivity test (LIST) using patient peripheral lymphocytes can predict the therapeutic efficacy of immunosuppressive drugs used in renal transplantation. We have evaluated the pharmacological efficacy of drugs by using the LIST with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, which measures the cellular mitochondrial activity. The LIST with the MTT assay requires a relatively large amount of blood. As such, we developed a new assay for examining drug sensitivity with a CellTiter-Glo assay, which measures the amount of cellular ATP to help increase the assay’s sensitivity and reduce the amount of blood needed. Renal transplant recipients generally receive either cyclosporine or tacrolimus, in addition to mycophenolate mofetil and methylprednisolone, as an immunosuppressive therapy to prevent acute rejection. We evaluated the pharmacological efficacy of these immunosuppressive agents with both the MTT and CellTiter-Glo assays using the peripheral blood mononuclear cells of 21 healthy volunteers. Furthermore, we also examined the relationship between these immunosuppressive agents’ pharmacological efficacy and the results of the MTT and CellTiter-Glo assays. The IC50 values for cyclosporine, tacrolimus, mycophenolic acid, and methylprednisolone were significantly correlated between the MTT and CellTiter-Glo assays. The amount of blood cells required for LIST with the CellTiter-Glo assay was able to be reduced to 25% of the amount required for the previously established LIST with the MTT assay procedure. We concluded from these observations that the LIST with the CellTiter-Glo assay should be used instead of the MTT assay for carrying out individualized immunosuppressive therapy in renal transplantation patients.

Key words: CellTiter-Glo assay; Lymphocyte immunosuppressant sensitivity test (LIST); 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; Peripheral blood mononuclear cells (PBMCs)

Received April 30, 2013; final acceptance October 7, 2013. Online prepub date: October 23, 2013.
Address correspondence to Hiroyasu Sasahara, Assistant Director, Division of Pharmacy, Niigata University Medical and Dental Hospital, 754 Asahimachi-dori 1-bancho, Chuo-ku, Niigata 951-8520, Japan. Tel: +81-25-227-2786; Fax; +81-25-227-0802; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 6, pp. 47–55, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674216
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Peripheral Lymphocyte Response to Mycophenolic Acid In Vitro and Incidence of Cytomegalovirus Infection in Renal Transplantation

Kentaro Sugiyama,* Hiroyasu Sasahara,† Mahoto Tsukaguchi,† Kazuya Isogai,† Akira Toyama,† Hiroshi Satoh,† Kazuhide Saitoh,‡ Yuki Nakagawa,‡ Kota Takahashi,‡ Sachiko Tanaka,* Kenji Onda,* and Toshihiko Hirano*

*Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Japan
†Division of Pharmacy, Niigata University Medical and Dental Hospital, Niigata, Japan
‡Division of Urology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan

The lymphocyte immunosuppressant sensitivity test (LIST) with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay procedure has been used to predict the pharmacological efficacy of immunosuppressive agents to prevent acute rejection episodes for renal transplant recipients. In this study, mycophenolic acid (MPA) pharmacological efficacies were evaluated by LIST at both prior to and just after renal transplantation. We compared the efficacies to the clinical outcome of these recipients. MPA’s pharmacological efficacy was evaluated by LIST not only before the operation but also at 2, 4, and 6 weeks after transplantation in 16 renal transplant recipients. These recipients were divided into high- and low-sensitivity groups according to peripheral blood mononuclear cell (PBMC) sensitivity to MPA in vitro. The MPA sensitivities were compared to cytomegalovirus (CMV) infection and acute rejection episodes in these recipients under MPA immunosuppressive therapy. The rate of CMV infection episodes in the low-MPA pharmacological efficacy group categorized at 2 weeks after renal transplantation was 5/6 (83.3%), which was significantly higher than the rate of 1/10 (10.0%) (p < 0.01) in the high-MPA sensitivity group. However, the MPA pharmacological efficacy evaluated both before and after transplantation had no relationship with the incidence of rejection episodes. These findings suggest that the MPA pharmacological efficacy evaluated by LIST at 2 weeks after operation is a useful biomarker for predicting the following occurrence of CMV infection episodes in renal transplant recipients.

Key words: Cytomegalovirus (CMV); Lymphocyte immunosuppressant sensitivity test (LIST); Mycophenolate mofetil (MMF); Mycophenolic acid (MPA); Peripheral blood mononuclear cell (PBMC); Renal transplantation

Received April 30, 2013; final acceptance October 4, 2013. Online prepub date: October 23, 2013.
Address correspondence to Kentaro Sugiyama, Associate Professor, Ph.D., Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji City, Tokyo 192-0392, Japan. Tel: +81-42-676-5111; Fax: +81-42-676-5798; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 6, pp. 57–62, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674252
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Experimental Nonalcoholic Steatohepatitis Induced by Neonatal Streptozotocin Injection and a High-Fat Diet in Rats

Huai-Che Hsu,*† Masaharu Dozen,* Naoto Matsuno,* Hiromichi Obara,‡ Ryou Tanaka,† 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
‡Department of Mechanical Engineering, Graduate School of Science and Engineering, Tokyo Metropolitan University, Tokyo, Japan

Nonalcoholic steatohepatitis (NASH) has become a major concern in clinical hepatology. To elucidate the disease mechanisms and to develop a treatment, the advent of an appropriate experimental model is crucial. Pregnant Sprague–Dawley rats were fed a high-fat diet from gestational day 16. Two days after birth, the neonates were injected subcutaneously with streptozotocin (STZ) (180, 200, or 256 mg/kg). The mothers were fed a high-fat diet during the nursing period. After being weaned (4 weeks of age), the juvenile rats were fed the same high-fat diet. The survival rates at the time of weaning were 25.6% (180 mg/kg STZ), 22.8% (200 mg/kg STZ), and 19.4% (256 mg/kg STZ). The mean body weight of NASH rats was approximately 20% less than that of normal rats. Serum levels of glucose, alanine aminotransferase, and hyaluronic acid increased in NASH rats. Histologically, typical features of steatohepatitis such as ballooning, inflammatory cell infiltration, and perivenular and pericellular fibrosis were observed. In an indocyanine green loading test, the blood half-life was significantly longer in NASH rats (5.04 ± 2.14 vs. 2.72 ± 0.72 min; p < 0.05), which was suggestive of an impaired hepatobiliary transportation function. Concomitantly, biliary ICG concentrations in NASH rats stabilized in a delayed fashion compared with normal rats. In addition, the amount of bile excreted in NASH rats was significantly lower than that in normal rats (4.32 ± 0.83 vs. 7.66 ± 1.05 mg/min; p < 0.01). The rat NASH model presented here mimics the clinical features of the disease and will be a helpful tool for medical and bioscience research.

Key words: Nonalcoholic steatohepatitis (NASH); Streptozotocin (STZ); High-fat diet; Rat

Received April 30, 2013; final acceptance October 10, 2013. Online prepub date: October 23, 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


Cell Medicine, Vol. 6, pp. 63–73, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674225
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Electron Therapy Attenuated Elevated Alanine Aminotransferase and Oxidative Stress Values in Type 2 Diabetes-Induced Nonalcoholic Steatohepatitis of Rats

Shin Enosawa,* Masaharu Dozen,*† 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

Chronic oxidative stress plays a key role in the progression of nonalcoholic steatohepatitis (NASH). We examined the efficacy of antioxidative electron treatment on type 2 diabetes-induced NASH in a rat model. We established NASH model rats, induced by neonatal administration of streptozotocin and a high-fat diet, which exhibited pathologically high values of alanine aminotransferase (ALT), glucose, and malondialdehyde (MDA). The rats were exposed to electron discharge at very low energy for 4 weeks; this dose results in the reduction of Fe3+ and glutathione disulfide in vitro. Serum ALT values were increased from baseline (8 weeks) to 125.0 ± 13 U/L at 10 weeks in the control group. In contrast, the values in the treated group did not show any increase at 10 weeks [87 ± 10 U/L (p = 0.0391)]. Hepatic MDA levels were also significantly decreased at 12 weeks (p < 0.05), but 8-hydroxy-2′-deoxyguanosine values were not statistically significant (p = 0.076). A gradual but steadily decreasing trend from initially high glucose levels was observed, though the values were not significant in 12-week-old animals (p = 0.074). However, the serum values of MDA, ALT, and glucose were well correlated. The progression of fibrosis as measured by increased serum levels of hyaluronic acid and histological examinations were not affected by the treatment in this model. Antioxidative electron treatment at very low energy attenuated the pathogenically elevated liver inflammation and oxidative stress, together with presumably impaired glucose metabolism in NASH rat model.

Key words: Diabetes mellitus; Type 2; Electrons; Hepatitis; Inflammation; Oxidative stress

Received April 30, 2013; final acceptance October 8, 2013. Online prepub date: October 23, 2013.
Address correspondence to Shin Enosawa, Division for Advanced Medical Sciences, Clinical Research Center, National Center for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-853, 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


Cell Medicine, Vol. 6, pp. 75–81, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674234
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

STO Feeder Cells Are Useful for Propagation of Primarily Cultured Human Deciduous Dental Pulp Cells by Eliminating Contaminating Bacteria and Promoting Cellular Outgrowth

Tomoya Murakami,* Issei Saitoh,* Emi Inada,† Mie Kurosawa,* Yoko Iwase,* Hirofumi Noguchi,‡ Yutaka Terao,§ Youichi Yamasaki,† Haruaki Hayasaki,* and Masahiro Sato¶

*Division of Pediatric Dentistry, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan
†Department of Pediatric Dentistry, Kagashima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan
‡Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
§Division of Microbiology and Infectious Diseases, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan
¶Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Kagoshima, Japan

STO feeder cells, a line established from mouse SIM embryonic fibroblasts, have been frequently used for establishing embryonic stem cells and maintaining them in an undifferentiated state. There are some reports demonstrating that fibroblastic cells have the ability to phagocytose Gram-positive bacterium (e.g., streptococci and staphylococci). In this study, we examined the possibility that STO cells could phagocytose Streptococcus mutans (a bacteria causing tooth decay), which always contaminates cultures of primarily isolated human deciduous dental pulp cells (HDDPCs). Simple cultivation of the primary HDDPCs in the absence of STO cells allowed S. mutans to massively propagate in the medium, thus leading to an opaque medium. In contrast, there was no bacterial contamination in the cultures containing mitomycin C (MMC)-inactivated STO cells. Furthermore, STO cells indicated bacterial phagocytic activity under fluorescent microscopy with the dye pHrodo. Besides removal of contaminating bacteria, STO feeder cells allowed the HDDPCs to spread out. These data suggest that MMC-treated STO cells can be useful for propagation of HDDPCs by eliminating contaminating bacteria and by promoting cellular outgrowth.

Key words: Feeder cells; STO cells; ES cells; Pluripotent stem cells; Phagocytosis; Primary tooth; Dental pulp cells

Received April 30, 2013; final acceptance October 31, 2013. Online prepub date: November 25, 2013.
Address correspondence to Issei Saitoh, D.D.S., Ph.D., Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University, 2-5274, Gakkocho-dori, Chuo-ku, Niigata, 951-8514, Japan. Tel: +81-25-227-2912; Fax: +81-25-227-2912; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 6, pp. 83–90, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674270
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Induced Pluripotent Stem Cell Labeling Using Quantum Dots

Hiroshi Yukawa,* Kaoru Suzuki,* Yuki Kano,† Tatsuya Yamada,† Noritada Kaji,*‡ Tetsuya Ishikawa,† and Yoshinobu Baba*‡§

*FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Japan
†Department of Medical Technology, Nagoya University, Graduate School of Medicine, Daikominami, Higashi-ku, Nagoya, Japan
‡Department of Applied Chemistry, Nagoya University, Graduate School of Engineering, Furo-cho, Chikusa-ku, Nagoya, Japan
§Health Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho, Takamatsu, Japan

Induced pluripotent stem (iPS) cells have received remarkable attention as the cell sources for clinical applications of regenerative medicine including stem cell therapy. Additionally, labeling technology is in high demand for tracing transplanted cells used in stem cell therapy. In this study, we used quantum dots (QDs), which have distinct fluorescence abilities in comparison with traditional probes, as the labeling materials and investigated whether iPS cells could be labeled with QDs with no cytotoxicity. iPS cells could not be labeled with QDs alone but required the use of cell-penetrating peptides such as octaarginine (R8). No significant cytotoxicity to iPS cells was confirmed by up to 8 nM QDs, and the iPS cells labeled with QDs maintained their undifferentiated state and pluripotency. These data suggest that QDs can be used for fluorescence labeling of iPS cells.

Key words: Induced pluripotent stem (iPS) cells; Quantum dots (QDs); Octaarginine (R8)

Received April 30, 2013; final acceptance October 23, 2013. Online prepub date: October 29, 2013.
Address correspondence to Hiroshi Yukawa, FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan. Tel: +81-52-789-5654; Fax: +81-52-789-3560; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 6, pp. 91–97, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517913X674261
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adipose Tissue-Derived Stem Cell Imaging Using Cadmium-Free Quantum Dots

Yoshiyuki Miyazaki,* Hiroshi Yukawa,† Hiroyasu Nishi,‡ Yukihiro Okamoto,† Noritada Kaji,*† Tsukasa Torimoto,‡ and Yoshinobu Baba*†§

*Department of Applied Chemistry, Nagoya University Graduate School of Engineering, Furo-cho, Chikusa-ku, Nagoya, Japan
†FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Japan
‡Department of Crystalline Materials Science, Nagoya University Graduate School of Engineering, Furo-cho, Chikusa-ku, Nagoya, Japan
§Health Technology Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho, Takamatsu, Japan

Quantum dots (QDs) have received much attention for biomolecule and cell imaging applications because of their superior optical properties such as high quantum efficiency, size-tunable emission, and resistance to photobleaching process. However, QDs that are commercially available contain cadmium (Cd), a highly toxic element. Thus, the development of Cd-free and less toxic QDs is strongly desired. In this study, we developed Cd-free QDs (ZnS-coated ZnS-AgInS2 solid solution nanoparticles with a sulfo group: ZnS-ZAIS-SO3H) and investigated the ability of this material to label stem cells. ZnS-ZAIS-SO3H could be transduced into mouse adipose tissue-derived stem cells (mASCs) using octaarginine peptides (R8), known as cell-penetrating peptides. The optimal ratio of ZnS-ZAIS-SO3H:R8 was found to be 1:100 for labeling mASCs. More than 80% of mASCs labeled with 500 nM ZnS-ZAIS-SO3H were found to be alive, and the proliferation rates of labeled mASCs were maintained at the same rate as that of nonlabeled mASCs. In addition, no abnormalities in the morphology of mASCs labeled with ZnS-ZAIS-SO3H could be observed. These data suggest that ZnS-ZAISSO3H may be effective for the labeling of mASCs.

Key words: Cadmium-free quantum dots (Cd-free QDs); Octaarginine (R8); Adipose tissue-derived stem cells (ASCs)

Received April 30, 2013; final acceptance October 23, 2013. Online prepub date: October 29, 2013.
Address correspondence to Hiroshi Yukawa, FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan. Tel: +81-52-789-5654; Fax: 81-52-789-5117; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it