Cell Transplantation 25(3) Abstracts

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Cell Transplantation, Vol. 25, pp. 425-436, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689523
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Injection of Dental Pulp Stem Cells Promotes Healing of Damaged Bladder Tissue in a Rat Model of Chemically Induced Cystitis

Yujiro Hirose,*†‡ Tokunori Yamamoto,* Misako Nakashima,†Yasuhito Funahashi,* Yoshihisa Matsukawa,* Masaya Yamaguchi,‡ Shigetada Kawabata,‡ and Momokazu Gotoh*

*Department of Urology, Nagoya University Graduate School of Medicine, Nagoya, Japan
†Department of Dental Regenerative Medicine, Center of Advanced Medicine for Dental and Oral Diseases, National Center for Geriatrics and Gerontology, Obu, Japan
‡Department of Oral and Molecular Microbiology, Osaka University Graduate School of Dentistry, Suita, Japan

Dental pulp stem cells (DPSCs) are reported as sources of mesenchymal stem cells (MSCs). MSCs are used as cell therapy options for various diseases. The present study examined the healing effects of DPSC injection on damaged bladder tissue in a chemically induced cystitis rat model. Cystitis was induced by hydrochloride injection into the bladder of female F344/NSlc rats. On the following day, DPSCs suspended in phosphate-buffered saline (PBS) were injected into the bladder and maintained for 1 h (DPSC injection group), while PBS alone was injected as the standard for comparison (PBS injection group). After 2 days following injection, considerable submucosal edema, vascular structure destruction, hemorrhage, and inflammatory cell invasion were observed both in the DPSC and PBS injection groups, with no difference in their degree of submucosal edema and hemorrhage. Six days after injection, vascular structure regeneration was observed in both groups; however, unlike the DPSC injection group, the PBS injection group showed traces of submucosal edema and hemorrhage. These results correlated with tissue concentrations of myeloperoxidase (MPO) and the inflammatory cytokines IL-1β, IL-6, and TNF-α. Furthermore, the intercontraction interval was prolonged, and the frequency of nociceptive behaviors was reduced in the DPSC injection group compared with the PBS injection group. DPSCs were found on the bladder epithelium until day 3 after injection. In the DPSC-conditioned media (CM), the trophic factors FGF-2, VEGF, and the C-C and C-X-C families of chemokines were detected. The results of DPSC injection into the cystitis rat model suggested that the injected cells promote the healing of the damaged bladder tissue by exerting various trophic effects while localizing on the bladder epithelium and that MSC injection is a potential novel therapy for interstitial cystitis/painful bladder syndrome.

Key words: Dental pulp stem cells (DPSCs); Cystitis; Cell injection; Trophic factor; Bladder epithelium

Received February 17, 2015; final acceptance October 26, 2015. Online prepub date: September 21, 2015.
Address correspondence to Yujiro Hirose, Department of Urology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. Tel: +81-52-744-2985; Fax: +81-52-744-2319; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 437-448, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689569
E-ISSN 1555-3892
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Outcome of Burns Treated With Autologous Cultured Proliferating Epidermal Cells: A Prospective Randomized Multicenter Intrapatient Comparative Trial

Kim L. M. Gardien,*† Roos E. Marck,‡ Monica C. T. Bloemen,‡ Taco Waaijman,§ Sue Gibbs,§¶# Magda M. W. Ulrich,†‡** Esther Middelkoop,*†‡ and Dutch Outback Study Group1

*Department of Plastic, Reconstructive and Hand Surgery, VU University Medical Center, Amsterdam, The Netherlands †MOVE Research Institute, Amsterdam, The Netherlands
‡Association of Dutch Burn CentresBeverwijk, The Netherlands
§Department of Dermatology, VU University Medical Center, Amsterdam, The Netherlands
¶A-SKIN BV, VU University Medical Center, Amsterdam, The Netherlands
#Department of Oral Cell Biology, Academic Center for Dentistry Amsterdam (ACTA), Amsterdam, The Netherlands
**Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands

Standard treatment for large burns is transplantation with meshed split skin autografts (SSGs). A disadvantage of this treatment is that healing is accompanied by scar formation. Application of autologous epidermal cells (keratinocytes and melanocytes) may be a suitable therapeutic alternative, since this may enhance wound closure and improve scar quality. A prospective, multicenter randomized clinical trial was performed in 40 adult patients with acute full thickness burns. On two comparable wound areas, conventional treatment with SSGs was compared to an experimental treatment consisting of SSGs in combination with cultured autologous epidermal cells (ECs) seeded in a collagen carrier. The primary outcome measure was wound closure after 5–7 days. Secondary outcomes were safety aspects and scar quality measured by graft take, scar score (POSAS), skin colorimeter (DermaSpectrometer®) and elasticity (Cutometer®). Wound epithelialization after 5–7 days was significantly better for the experimental treatment (71%) compared to the standard treatment (67%) (p = 0.034, Wilcoxon), whereas the take rates of the grafts were similar. No related adverse events were recorded. Scar quality was evaluated at 3 (n= 33) and 12 (n = 28) months. The POSAS of the observer after 3 and 12 months and of the patient after 12 months were significantly better for the experimental area. Improvements between 12% and 23% (p ≤ 0.010, Wilcoxon) were detected for redness, pigmentation, thickness, relief, and pliability. Melanin index at 3 and 12 months and erythema index at 12 months were closer to normal skin for the experimental treatment than for conventional treatment (p ≤ 0.025 paired samples t-test). Skin elasticity showed significantly higher elasticity (p = 0.030) in the experimental area at 3 months follow-up. We showed a safe application and significant improvements of wound healing and scar quality in burn patients after treatment with ECs versus SSGs only. The relevance of cultured autologous cells in treatment of extensive burns is supported by our current findings.

Key words: Advanced therapy medicinal products (ATMPs); Autologous keratinocytes; Epithelial cells; Burns; Scar quality; Wound healing

Received November 4, 2014; final acceptance October 27, 2015. Online prepub date: September 28, 2015.
1See Acknowledgments for the complete list of authors from the Dutch Outback Study Group.
Address correspondence to Esther Middelkoop, VU University Medical Center, Department of Plastic, Reconstructive and Hand Surgery, PO Box 7057, 1007 MB Amsterdam, The Netherlands. Tel: 0031251275500; Fax: 0031251216059; 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 Transplantation, Vol. 25, pp. 449-461, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688641
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Fetal Cartilage-Derived Cells Have Stem Cell Properties and Are a Highly Potent Cell Source for Cartilage Regeneration

Woo Hee Choi,* Hwal Ran Kim,* Su Jeong Lee,* Nayoung Jeong,* So Ra Park,† Byung Hyune Choi,‡ and Byoung-Hyun Min*§¶

*Department of Molecular Science and Technology, Ajou University, Suwon, Korea
†Department of Physiology, Inha University College of Medicine, Incheon, Korea
‡Department of Biomedical Sciences, Inha University College of Medicine, Incheon, Korea
§Cell Therapy Center, Ajou University Medical Center, Suwon, Korea
¶Department of Orthopedic Surgery, Ajou University School of Medicine, Suwon, Korea

Current strategies for cartilage cell therapy are mostly based on the use of autologous chondrocytes or mesenchymal stem cells (MSCs). However, these cells have limitations of a small number of cells available and of low chondrogenic ability, respectively. Many studies now suggest that fetal stem cells are more plastic than adult stem cells and can therefore more efficiently differentiate into target tissues. However, the characteristics and the potential of progenitor cells from fetal tissue remain poorly defined. In this study, we examined cells from human fetal cartilage at 12 weeks after gestation in comparison with bone marrow-derived MSCs or cartilage chondrocytes from young donors (8–25 years old). The fetal cartilage-derived progenitor cells (FCPCs) showed higher yields by approximately 24 times than that of chondrocytes from young cartilage. The morphology of the FCPCs was polygonal at passage 0, being similar to that of the young chondrocytes, but it changed later at passage 5, assuming a fibroblastic shape more akin to that of MSCs. As the passages advanced, the FCPCs showed a much greater proliferation ability than the young chondrocytes and MSCs, with the doubling times ranging from 2~4 days until passage 15. The surface marker profile of the FCPCs at passage 2 was quite similar to that of the MSCs, showing high expressions of CD29, CD90, CD105, and Stro-1. When compared to the young chondrocytes, the FCPCs showed much less staining of SA-β-gal, a senescence indicator, at passage 10 and no decrease in SOX9 expression until passage 5. They also showed a much greater chondrogenic potential than the young chondrocytes and the MSCs in a three-dimensional pellet culture in vitro and in polyglycolic acid (PGA) scaffolds in vivo. In addition, they could differentiate intoadipogenic and osteogenic lineages as efficiently as MSCs in vitro. These results suggest that FCPCs have stem cell properties to some extent and that they are more active in terms of proliferation andchondrogenic differentiation than young chondrocytes or MSCs.

Key words: Fetal cartilage-derived progenitor cells (FCPCs); Mesenchymal stem cells (MSCs); Chondrocytes; Cartilage regeneration

Received January 18, 2014; final acceptance July 28, 2015. Online prepub date: July 13, 2015.
Address correspondence to Professor Byoung-Hyun Min, Department of Orthopedic Surgery, Ajou University School of Medicine, Wonchon-dong, Youngtong-gu, Suwon, Gyeonggi-do, Korea. Tel: +82-31-219-4444; Fax: +82-31-219-4193; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Professor Byung Hyune Choi, Department of Biomedical Sciences, Inha University College of Medicine, 100 Inharo, Nam-gu, Incheon, 22212, Korea. Tel: +82-32-860-9881; Fax: +82-32-885-8302; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 463-479, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688515
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Human Fetal Progenitor Tenocytes for Regenerative Medicine

A. Grognuz,* C. Scaletta,* A. Farron,† W. Raffoul,* and L. A. Applegate*

*Unit of Regenerative Therapy, Service of Plastic, Reconstructive and Hand Surgery, Department of Musculoskeletal Medicine, University Hospital of Lausanne, Switzerland
†Service of Orthopaedics and Traumatology, Department of Musculoskeletal Medicine, University Hospital of Lausanne, Switzerland

Tendon injuries are very frequent and affect a wide and heterogeneous population. Unfortunately, the healing process is long with outcomes that are not often satisfactory due to fibrotic tissue appearance, which leads to scar and adhesion development. Tissue engineering and cell therapies emerge as interesting alternatives to classical treatments. In this study, we evaluated human fetal progenitor tenocytes (hFPTs) as a potential cell source for treatment of tendon afflictions, as fetal cells are known to promote healing in a scarless regenerative process. hFPTs presented a rapid and stable growth up to passage 9, allowing to create a large cell bank for off-the-shelf availability. hFPTs showed a strong tenogenic phenotype with an excellent stability, even when placed in conditions normally inducing cells to differentiate. The karyotype also indicated a good stability up to passage 12, which is far beyond that necessary for clinical application (passage 6). When placed in coculturehFPTs had the capacity to stimulate human adult tenocytes (hATs), which are responsible for the deposition of a new extracellular matrix during tendon healing. Finally, it was possible to distribute cells in porous or gel scaffolds with an excellent survival, thus permitting a large variety of applications (from simple injections to grafts acting as filling material). All of these results are encouraging in the development of an off-the-shelf cell source capable of stimulating tendon regeneration for the treatment of tendon injuries.

Key words: Cell banking; Fetal cell therapy; Tendon healing; Tendon injuries; Tenocytes

Received April 23, 2015; final acceptance September 3, 2015. Online prepub date: June 24, 2015.
Address correspondence to Professor Lee Ann Laurent-Applegate, University Hospital of Lausanne (CHUV), Service of Plastic, Reconstructive and Hand Surgery, Regenerative Therapy Unit (UTR), EPCR/Croisettes 22, CH-1066 Epalinges, Switzerland. Tel: + 41 21 314 35 10; Fax: + 41 21 887 84 14; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 481-489, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688902
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Extensive Loss of Islet Mass Beyond the First Day After Intraportal Human Islet Transplantation in a Mouse Model

Hanna Liljebäck,*Liza Grapensparr,* Johan Olerud,† and Per-Ola Carlsson*‡

*Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden
†Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
‡Department of Medical Sciences, Uppsala University, Uppsala, Sweden

Clinical islet transplantation is characterized by a progressive deterioration of islet graft function, which renders many patients once again dependent on exogenous insulin administration within a couple of years. In this study, we aimed to investigate possible engraftment factors limiting the survival and viability of experimentally transplanted human islets beyond the first day after their transplantation to the liver. Human islets were transplanted into the liver of nude mice and characterized 1 or 30 days after transplantation by immunohistochemistry. The factors assessed were endocrine mass, cellular death, hypoxia, vascular density and amyloid formation in the transplanted islets. One day posttransplantation, necrotic cells, as well as apoptotic cells, were commonly observed. In contrast to necrotic death, apoptosis rates remained high 1 month posttransplantation, and the total islet mass was reduced by more than 50% between 1 and 30 days posttransplantation. Islet mass at 30 days posttransplantationcorrelated negatively to apoptotic death. Vascular density within the transplanted islets remained less than 30% of that in native human islets up to 30 days posttransplantation and was associated with prevailing hypoxia. Amyloid formation was rarely observed in the 1-day-old transplants, but was commonly observed in the 30-day-old islet transplants. We conclude that substantial islet cell death occurs beyond the immediate posttransplantation phase, particularly through apoptotic events. Concomitant low vascularization with prevailing hypoxia and progressive amyloid development was observed in the human islet grafts. Strategies to improve engraftment at the intraportal site or change of implantation site in the clinical setting are needed.

Key words: Islet transplantation; Diabetes; Amyloid; Engraftment

Received October 24, 2014; final acceptance August 18, 2015. Online prepub date: August 10, 2015.
Address correspondence to Per-Ola Carlsson, M.D., Ph.D., Department of Medical Cell Biology, University of Uppsala, Husargatan 3, Box 571, SE-75123 Uppsala, Sweden. Tel: +46 18 4714425; Fax: +46 18 4714059; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 491-501, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688498
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Islet Volume and Indexes of β-Cell Function in Humans

Rita Nano,* Raffaella Melzi,* Alessia Mercalli,* Gianpaolo Balzano,† Marina Scavini,* Riccardo Bonadonna,‡§ and Lorenzo Piemonti*

*Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy
†Pancreatic Surgery Unit, IRCCS San Raffaele Scientific Institute, Milan, Italy
‡Division of Endocrinology, Ospedale Maggiore, Parma, Italy
§University of Parma School of Medicine, Parma, Italy

Islet volume and endocrine pancreas architecture (islet size distribution) may be independent determinants of β-cell function. Furthermore, the accuracy of homeostatic model assessment (HOMA) indexes in predicting β-cell mass has never been assessed. Here we investigated the relationships between islet volume, islet density, and islet size distribution, estimated after pancreatic tissue digestion, with established indexes of β-cell function in humans. We included in this study 42 patients who were candidates for islet autotransplantation and had well-characterized glucose metabolism. Indexes of insulin secretion were calculated and compared with the islet volume, as a surrogate of β-cell mass, obtained after digestion of pancreas. Islet counting analysis showed considerable interindividual variation in islet density and size. Islet volume, but not density nor size, was the only independent determinant of β-cell function assessed by insulin HOMA β-cell. Islet volume was significantly reduced in the patients with overt hyperglycemia, but not in patients with impaired fasting glucose. Insulin HOMA β-cell predicted islet volume better than other measures of fasting insulin secretion. In conclusion, the present study documented a close direct relationship between indexes of β-cell function and islet volume in humans. The insulin HOMA β-cell provides a more reliable estimate of pancreatic islet volume than fasting glucose before islet isolation.

Key words: Human islets; Homeostatic model assessment (HOMA); Islet density; Islet volume; Islet size; β-Cell mass

Received April 3, 2015; final acceptance June 19, 2015. Online prepub date: June 22, 2015.
Address correspondence to Lorenzo Piemonti, San Raffaele Scientific Institute, Via Olgettina 60, 20132 Milan, Italy. Tel: +39 02 26432706; Fax: +39 02 26432871; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 503-515, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688461
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Characterization of Innate Immunity in an Extended Whole Blood Model of Human Islet Allotransplantation

Maria Hårdstedt,*† Susanne Lindblom,* Alex Karlsson-Parra,* Bo Nilsson,* and Olle Korsgren*

*Department of Immunology, Genetics and Pathology, Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden
†Center for Clinical Research Dalarna-Uppsala University, Falun, Sweden

The instant blood-mediated inflammatory reaction (IBMIR) has been studied in whole blood models of human allo-islet transplantation for short periods (<6 h). Beyond this time frame the innate response tointraportally transplanted islets is less well described. A novel whole blood model was applied to study blood–islet–graft interactions up to 48 h. Heparinized polyvinyl chloride tubing was sealed into small bags containing venous blood together with allogeneic human islets and exocrine tissue, respectively. The bags were attached to a rotating wheel (37°C). Concentrated glucose and sodium hydrogen carbonate were added every 12 h to maintain physiological limits for sustained immune cell functions. Plasma was collected at repeated time points for analyses of coagulation/complement activation and chemokine/cytokine production. Immune cell infiltration was analyzed using immunohistochemistry. Coagulation and platelet activation markers, thrombin–antithrombin complex (TAT) and soluble CD40 ligand (sCD40L) showed early high concentrations (at 6–12 h). sC5b-9 steadily increased over 48 h. At 6 h neutrophils and monocytes surrounded the clotted cellular grafts with a following massive infiltration of neutrophils. High and increasing concentrations of CXCR1/2 ligands [IL-8 and growth-regulated oncogene α/β/γ (Gro- α/β/γ)] and IL-6 were produced in response to human islets and exocrine tissue. The CCR2 ligand monocyte chemoattractant protein 1 (MCP-1) exhibited increasing concentrations in response to exocrine tissue. The CXCR3 ligand interferon-inducible T cell a chemoattractant (I-TAC) was produced in response to both human islets and exocrine tissue from 6 h. Monokine induced by γ interferon (Mig) and interferon γ-induced protein 10 (IP-10) showed a later response, preferentially to exocrine tissue and with larger variations among preparations. An extended blood model of clinical islet transplantation allowed characterization of early immune activation in response to human islets and exocrine tissue. Increased production of chemokines targeting CXCR1/2, CCR2, and CXCR3 was observed, accompanied by massive intraislet neutrophil infiltration over 48 h. The model proved to be useful in exploring early blood-mediated reactions to cellular transplants and has relevance for evaluation of pharmacological interventions to prevent graft loss.

Key words: Chemokines; Instant blood-mediated inflammatory reaction (IBMIR); Innate immunity; Islet transplantation; Whole blood model

Received April 13, 2015; final acceptance July 16, 2015. Online prepub date: June 16, 2015.
Address correspondence to Maria Hårdstedt, Center for Clinical Research Dalarna, Nissers väg 3, SE-791 82 Falun, Sweden. Tel: +46-23-18311; Fax: +46-23-18375; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 517-530, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688542
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Induced Pluripotent Stem Cell-Derived Conditioned Medium Attenuates Acute Kidney Injury by Downregulating the Oxidative Stress-Related Pathway in Ischemia–Reperfusion Rats

Der-Cherng Tarng,*†‡§ Wei-Cheng Tseng,†‡¶ Pei-Ying Lee,*# Shih-Hwa Chiou,**†† and Shie-Liang Hsieh†‡‡

*Department and Institute of Physiology, National Yang-Ming University, Taipei, Taiwan
†Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
‡School of Medicine, National Yang-Ming University, Taipei, Taiwan
§Division of Nephrology, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan
¶Division of Nephrology, Department of Medicine, Taipei City Hospital Heping Fuyou Branch, Taipei, Taiwan
#General Education Center, Tzu Chi College of Technology, Hualien, Taiwan
**Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan ††Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan
‡‡Genomics Research Center, Academia Sinica, Taipei, Taiwan

Teratoma-like formation addresses a critical safety concern for the potential utility of induced pluripotent stem cells (iPSCs). Therefore, therapy utilizing iPSC-derived conditioned medium (iPSC-CM) for acute kidney injury (AKI) has attracted substantial interest. A recent study showed that iPSC-CM effectively alleviated ventilator-induced lung injury in rats. It prompts us to assess the therapeutic effects of iPSC-CM on ischemic AKI. First, we assessed the changes in renal function and tubular cell apoptosis by intraperitoneal administration of iPSC-CM to ischemia–reperfusion (I/R) rats. Second, we explored the oxidative stress-related pathway in the apoptosis of renal tubular cells subjected to hypoxia–reoxygenation (H/R). Administration of iPSC-CM significantly improved renal function and protected tubular cells against apoptosis in rats with I/R-AKI, and the optimal effect was observed at the 50-fold concentrated iPSC-CM. iPSC-CM also mitigated the H/R-induced apoptosis of NRK-52E cells in vitro. Reactive oxygen species (ROS) production was augmented in kidneys following I/R and in NRK-52E cells subjected to H/R. Meanwhile, expressions of phosphorylated p38 MAPK, TNF-α, and cleaved caspase 3 and NF-κB activity were consistently increased in vivo and in vitro. Following administration of iPSC-CM, ROS production was abolished, and inflammatory cytokine expression was significantly suppressed. Annexin V–propidium iodide flow cytometry and in situ TUNEL assay further showed that iPSC-CM markedly attenuated H/R- or I/R-induced tubular cell apoptosis. Intriguingly, treatment with iPSC-CM significantly improved the survival of rats with I/R-induced AKI. iPSC-CM represents a favorable source of stem cell-based therapy and may serve as a potential therapeutic strategy for kidney repair in ischemic AKI.

Key words: Acute kidney injury (AKI); Conditioned medium; Hypoxia–reoxygenation (H/R); Induced pluripotent stem cells (iPSCs); Ischemia–reperfusion (I/R)

Received January 10, 2013; final acceptance September 5, 2015. Online prepub date: June 30, 2015.
Address correspondence to Professor Der-Cherng Tarng, M.D., Ph.D., Chief, Division of Nephrology, Department of Medicine, Taipei Veterans General Hospital, 201, Sec. 2, Shih-Pai Road, Taipei 11217, Taiwan. Tel: +886-2-2871 2121, ext. 2678; Fax: +886-2-2875 7824; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 531-538, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688623
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Pancreatic ʟ-Glutamine Administration Protects Pig Islets From Cold Ischemic Injury and Increases Resistance Toward Inflammatory Mediators

Heide Brandhorst,*† Bastian Theisinger,‡ Bernhard Guenther,‡ Paul R. Johnson,*†§ and Daniel Brandhorst*†

*Islet Transplant Research Group, Nuffield Department of Surgical Sciences, University of Oxford, Oxford, UK
†Oxford Centre for Diabetes, Endocrinology, and Metabolism (OCDEM), Churchill Hospital, Oxford, UK
Novaliq GmbH, Heidelberg, Germany
§Oxford NIHR Biomedical Research Centre, Oxford, UK

The isolation and transplantation of porcine islets represent a future option for the treatment of type 1 diabetic patients. Stringent product release criteria and limited availability of transgenic and specific pathogen-free pigs will essentially require processing of explanted pig pancreata in specialized, possibly remote isolation facilities, whereby pancreata are exposed to cold ischemia due to prolonged tissue transit time. In the present study we investigated whether pancreas oxygenation can be efficiently combined with an antioxidant strategy utilizing intraductal ʟ-glutamine administration. Pig pancreata wereintraductally perfused after retrieval and after cold storage in oxygen-precharged perfluorohexyloctane utilizing University of Wisconsin solution supplemented with (n = 16) or without (n = 14) 5 mmol/L ʟ-glutamine. After isolation purified islets were subjected to extensive quality assessment. Islet recovery postpurification was significantly higher in glutamine-treated pancreata (77.0±3.3% vs. 60.3±6.0%,p<0.05). Glutamine administration increased intraislet content of reduced glutathione (117.8±16.5 vs. 15.9±2.8 ng/ng protein, p><0.001) associated with increased islet recovery after culture (65.8±12.1% vs. 40.3±11.7%, p><0.05), enhanced glucose stimulation index (1.82±0.16 vs. 1.38±0.10, p><0.05), and improved posttransplant function in diabetic nude mice (p><0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and highdose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal l-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of l-glutamine as supplement for pancreas perfusion during organ procurement. Key words: Pig pancreas; Ischemia; Cold storage; Oxygenation; l-Glutamine> < 0.05). Glutamine administration increased intraislet content of reduced glutathione (117.8±16.5 vs. 15.9±2.8 ng/ng protein, p<0.001) associated with increased islet recovery after culture (65.8±12.1% vs. 40.3±11.7%, p><0.05), enhanced glucose stimulation index (1.82±0.16 vs. 1.38±0.10, p><0.05), and improved posttransplant function in diabetic nude mice (p><0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and highdose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal l-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of l-glutamine as supplement for pancreas perfusion during organ procurement. Key words: Pig pancreas; Ischemia; Cold storage; Oxygenation; l-Glutamine> < 0.001) associated with increased islet recovery after culture (65.8±12.1% vs. 40.3±11.7%, p<0.05), enhanced glucose stimulation index (1.82±0.16 vs. 1.38±0.10, p><0.05), and improved posttransplant function in diabetic nude mice (p><0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and highdose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal l-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of l-glutamine as supplement for pancreas perfusion during organ procurement. Key words: Pig pancreas; Ischemia; Cold storage; Oxygenation; l-Glutamine> < 0.05), enhanced glucose stimulation index (1.82±0.16 vs. 1.38±0.10, p<0.05), and improved posttransplant function in diabetic nude mice (p><0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and highdose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal l-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of l-glutamine as supplement for pancreas perfusion during organ procurement. Key words: Pig pancreas; Ischemia; Cold storage; Oxygenation; l-Glutamine> < 0.05), and improved posttransplant function in diabetic nude mice (p<0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and highdose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal l-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of l-glutamine as supplement for pancreas perfusion during organ procurement. Key words: Pig pancreas; Ischemia; Cold storage; Oxygenation; l-Glutamine> < 0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and highdose proinflammatorycytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal ʟ-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of ʟ-glutamine as supplement for pancreas perfusion during organ procurement.

Key words: Pig pancreas; Ischemia; Cold storage; Oxygenation; ʟ-Glutamine

Received April 2, 2015; final acceptance July 15, 2015. Online prepub date: July 8, 2015.
Address correspondence to Daniel Brandhorst, Nuffield Department of Surgical Sciences, Oxford Center for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Drive, Oxford OX3 7LE, UK. Tel: +44-1865-8-57252; Fax: +44-1865-8-57299; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 539-547, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689451
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Optimization and Scale-up Isolation and Culture of Neonatal Porcine Islets: Potential for Clinical Application

Cara Ellis,*† James G. Lyon,† and Gregory S. Korbutt*†

*Department of Surgery, University of Alberta, Edmonton, Canada
†Alberta Diabetes Institute, University of Alberta, Edmonton, Canada

One challenge that must be overcome to allow transplantation of neonatal porcine islets (NPIs) to become a clinical reality is defining a reproducible and scalable protocol for the efficient preparation of therapeutic quantities of clinical grade NPIs. In our standard protocol, we routinely isolate NPIs from a maximum of four pancreases, requiring tissue culture in 16 Petri dishes (four per pancreas) in Ham’s F10 and bovine serum albumin (BSA). We have now developed a scalable and technically simpler protocol that allows us to isolate NPIs from a minimum of 12 pancreases at a time by employing automated tissue chopping, collagenase digestion in a single vessel, and tissue culture/media changes in 75% fewer Petri dishes. For culture, BSA is replaced with human serum albumin and supplemented with Z-VAD-FMK general caspase inhibitor and a protease inhibitor cocktail. The caspase inhibitor was added to the media for only the first 90 min of culture. NPIs isolated using the scalable protocol had significantly more cellular insulin recovered (56.9 ± 1.4 µg) when compared to the standard protocol (15.0 ± 0.5 µg; p<0.05). Compared to our standard protocol, recovery of b-cells (6.0×106±0.2 vs. 10.0×106±0.4; p><0.05) and islet equivalents (35,135±186 vs. 41,810±226; p><0.05) was significantly higher using the scalable protocol. During a static glucose stimulation assay, the SI of islets isolated by the standard protocol were significantly lower than the scale-up protocol (4.3±0.2 vs. 5.5±0.1; p><0.05). Mice transplanted with NPIs using the scalable protocol had significantly lower blood glucose levels than the mice that receiving NPIs from the standard protocol (p><0.01) and responded significantly better to a glucose tolerance test. Based on the above findings, this improved simpler scalable protocol is a significantly more efficient means for preparing therapeutic quantities of clinical grade NPIs. Key words: Neonatal porcine islets (NPIs); Islet transplantation; Islet isolation> < 0.05). Compared to our standard protocol, recovery of b-cells (6.0 × 106 ± 0.2 vs. 10.0 × 106 ± 0.4; p<0.05) and islet equivalents (35,135±186 vs. 41,810±226; p><0.05) was significantly higher using the scalable protocol. During a static glucose stimulation assay, the SI of islets isolated by the standard protocol were significantly lower than the scale-up protocol (4.3±0.2 vs. 5.5±0.1; p><0.05). Mice transplanted with NPIs using the scalable protocol had significantly lower blood glucose levels than the mice that receiving NPIs from the standard protocol (p><0.01) and responded significantly better to a glucose tolerance test. Based on the above findings, this improved simpler scalable protocol is a significantly more efficient means for preparing therapeutic quantities of clinical grade NPIs. Key words: Neonatal porcine islets (NPIs); Islet transplantation; Islet isolation> < 0.05) and islet equivalents (35,135 ± 186 vs. 41,810 ± 226; p<0.05) > < 0.05) was significantly higher using the scalable protocol. During a static glucose stimulation assay, the SI of islets isolated by the standard protocol were significantly lower than the scale-up protocol (4.3 ± 0.2 vs. 5.5 ± 0.1; p<0.05). Mice transplanted with NPIs using the scalable protocol had significantly lower blood glucose levels than the mice that receiving NPIs from the standard protocol (p><0.01) and responded significantly better to a glucose tolerance test. Based on the above findings, this improved simpler scalable protocol is a significantly more efficient means for preparing therapeutic quantities of clinical grade NPIs.> < 0.05). Mice transplanted with NPIs using the scalable protocol had significantly lower blood glucose levels than the mice that receiving NPIs from the standard protocol (p<0.01) and responded significantly better to a glucose tolerance test. Based on the above findings, this improved simpler scalable protocol is a significantly more efficient means for preparing therapeutic quantities of clinical grade NPIs. Key words: Neonatal porcine islets (NPIs); Islet transplantation; Islet isolation> < 0.01) and responded significantly better to a glucose tolerance test. Based on the above findings, this improved simpler scalable protocol is a significantly more efficient means for preparing therapeutic quantities of clinical grade NPIs.

Key words: Neonatal porcine islets (NPIs); Islet transplantation; Islet isolation

Received May 19, 2015; final acceptance October 22, 2015. Online prepub date: September 15, 2015.
Address correspondence to Gregory S. Korbutt, Ph.D., 5-002 Li Ka Shing Centre for Health Research Innovation, Edmonton, AB, T6G 2E1, Canada. Tel: +1-780-492-4657; Fax: +1-780-492-5501; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it 539


Cell Transplantation, Vol. 25, pp. 549-558, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688669
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Efficacy of Multilayered Hepatocyte Sheet Transplantation for Radiation‑Induced Liver Damage and Partial Hepatectomy in a Rat Model

Zhassulan Baimakhanov, Kosho Yamanouchi, Yusuke Sakai, Makiko Koike, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Fumihiko Fujita, Kengo Kanetaka, Tamotsu Kuroki, and Susumu Eguchi

Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Sakamoto, Nagasaki, Japan

Although cell sheet technology has recently been developed for use in both animal experiments and in the clinical setting, it remains unclear whether transplanted hepatocyte sheets improve the liver function in vivo. Radiation-induced liver damage (RILD) combined with partial hepatectomy (PH) has been reported to suppress the proliferation of host hepatocytes and induce critical liver failure. The aim of this study was to improve the liver function in the above-mentioned diseased rat model (RILD + PH) using multilayered hepatocyte sheet transplantation. In this study, we used Fischer rats as a donor for primary hepatocytes and dermal fibroblast isolation. Cocultured multilayered hepatocyte sheets were generated by disseminating hepatocytes onto fibroblasts cultured beforehand on temperature-responsive culture dishes. Four cell sheets were transplanted into the recipient rats subcutaneously. Prior to transplantation, RILD (50 Gy) with 2/3PH was induced in the recipients. The same model was applied in the control group without transplantation. The serum was collected each week. The rats in both groups were sacrificed at 2 months after transplantation for the histological analysis. Consequently, the serum albumin concentrations were significantly higher in the transplant group than in the control group (54.3 ± 9.6 vs. 32.7 ± 5.7 mg/ml; p<0.01) after 2 months and comparable to the serum albumin levels in the normal rats (58.1±6.4 mg/ml). In addition, treatment with the transplanted sheets significantly improved the survival rate (57% vs. 22%, p><0.05), and the hepatocyte sheets showed the storage of albumin, glycogen, and bile canaliculus structures. Some hepatocytes and fibroblasts were positive for Ki-67, and vascularization was observed around the cell sheets. Transplanted multilayered hepatocyte sheets can survive with additional proliferative activity, thereby maintaining the liver function in vivo for at least 2 months, providing metabolic support for rats with RILD. Key words: Hepatocytes; Cell sheet; Transplantation; Rat diseased model; Metabolic support> < 0.01) after 2 months and comparable to the serum albumin levels in the normal rats (58.1 ± 6.4 mg/ml). In addition, treatment with the transplanted sheets significantly improved the survival rate (57% vs. 22%, p<0.05), and the hepatocyte sheets showed the storage of albumin, glycogen, and bile canaliculus structures. Some hepatocytes and fibroblasts were positive for Ki-67, and vascularization was observed around the cell sheets. Transplanted multilayered hepatocyte sheets can survive with additional proliferative activity, thereby maintaining the liver function in vivo for at least 2 months, providing metabolic support for rats with RILD. Key words: Hepatocytes; Cell sheet; Transplantation; Rat diseased model; Metabolic support> < 0.05), and the hepatocyte sheets showed the storage of albumin, glycogen, and bile canaliculus structures. Some hepatocytes and fibroblasts were positive for Ki-67, and vascularization was observed around the cell sheets. Transplanted multilayered hepatocyte sheets can survive with additional proliferative activity, thereby maintaining the liver function in vivo for at least 2 months, providing metabolic support for rats with RILD.

Key words: Hepatocytes; Cell sheet; Transplantation; Rat diseased model; Metabolic support

Received May 15, 2015; final acceptance September 15, 2015. Online prepub date: July 28, 2015.
Address correspondence to Susumu Eguchi, M.D., Ph.D., Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan. Tel: +81-95-819-7316; Fax: +81-95-819-7319; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 559-574, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688920
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Effect of an Epineurial-Like Biohybrid Nerve Conduit on Nerve Regeneration

Shu-Chih Hsieh,*† Chen-Jung Chang,‡ Wen-Tung Cheng,* Ting-Chen Tseng,§ and Shan-hui Hsu§

*Department of Chemical Engineering, National Chung Hsing University, Taichung, Taiwan
†Department of Chemical and Materials Engineering, National Chin-Yi University of Technology, Taichung, Taiwan
‡Department of Medical Imaging and Radiological Science, Central Taiwan University of Science and Technology, Taichung, Taiwan
§Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan

A novel approach of making a biomimetic nerve conduit was established by seeding adipose-derived adult stem cells (ADSCs) on the external wall of porous poly(D,L-lactic acid) (PLA) nerve conduits. The PLA conduits were fabricated using gas foaming salt and solvent–nonsolvent phase conversion. We examined the effect of two different porous structures (GS and GL) on ADSC growth and proliferation. The GS conduits had better structural stability, permeability, and porosity, as well as better cell viability at 4, 7, and 10 days. The epineuriallike tissue was grown from ADSC-seeded conduits cultured for 7 days in vitro and then implanted into 10-mm rat sciatic nerve defects for evaluation. The regeneration capacity and functional recovery were evaluated by histological staining, electrophysiology, walking track, and functional gait analysis after 6 weeks of implantation. Experimental data indicated that the autograft and ADSC-seeded GS conduits had better functional recovery than the blank conduits and ADSC-seeded GL conduits. The area of regenerated nerve and number of myelinated axons quantified based on the histology also indicated that the autograft and AGS groups performed better than the other two groups. We suggested that ADSCs may interact with endogenous Schwann cells and release neurotrophic factors to promote peripheral nerve regeneration. The design of the conduit may be critical for producing a biohybrid nerve conduit and to provide an epineurial-like support.

Key words: Adipose-derived adult stem cells (ADSCs); Poly(D,L-lactic acid) (PLA); Peripheral nerve regeneration

Received April 9, 2015; final acceptance September 22, 2015. Online prepub date: August 21, 2015.
Address correspondence to Shan-hui Hsu, Ph.D., Institute of Polymer Science and Engineering, National Taiwan University, No. 1, Sec. 4 Roosevelt Road, Taipei 10617, Taiwan, R.O.C. Tel: +886-2-33665313; Fax: +886-2-33665237; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 575-592, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688614
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

In Vivo Tracking of Human Neural Progenitor Cells in the Rat Brain Using Magnetic Resonance Imaging Is Not Enhanced by Ferritin Expression

Ksenija Bernau,*1 Christina M. Lewis,† Anna M. Petelinsek,‡ Matthew S. Reagan,* David J. Niles,† Virginia B. Mattis,§ M. Elizabeth Meyerand,*† Masatoshi Suzuki,‡ and Clive N. Svendsen§

*University of Wisconsin-Madison, Department of Biomedical Engineering, Madison, WI, USA
†University of Wisconsin-Madison, Medical Physics Department, Madison, WI, USA
‡University of Wisconsin-Madison, Department of Comparative Biosciences, Madison, WI, USA
§Cedars-Sinai Medical Center, Board of Governors Regenerative Medicine Institute, Los Angeles, CA, USA

Rapid growth in the field of stem cell research has generated a lot of interest in their therapeutic use, especially in the treatment of neurodegenerative diseases. Specifically, human neural progenitor cells (hNPCs), unique in their capability to differentiate into cells of the neural lineage, have been widely investigated due to their ability to survive, thrive, and migrate toward injured tissues. Still, one of the major roadblocks for clinical applicability arises from the inability to monitor these cells following transplantation. Molecular imaging techniques, such as magnetic resonance imaging (MRI), have been explored to assess hNPC transplant location, migration, and survival. Here we investigated whether inducing hNPCs to overexpress ferritin (hNPCsFer), an iron storage protein, is sufficient to track these cells long term in the rat striatum using MRI. We found that increased hypointensity on MRI images could establish hNPCFer location. Unexpectedly, however, wild-type hNPC transplants were detected in a similar manner, which is likely due to increased iron accumulation following transplantation-induced damage. Hence, we labeled hNPCs with superparamagnetic iron oxide (SPIO) nanoparticles to further increase iron content in an attempt to enhance cell contrast in MRI. SPIO-labeling of hNPCs (hNPCs-SPIO) achieved increased hypointensity, with significantly greater area of decreased T2* compared tohNPCFer (p<0.0001) and all other controls used. However, none of the techniques could be used to determine graft rejection in vivo, which is imperative for understanding cell behavior following transplantation. We conclude that in order for cell survival to be monitored in preclinical and clinical settings, another molecular imaging technique must be employed, including perhaps multimodal imaging, which would utilize MRI along with another imaging modality. Key words: Neural progenitor cells; Cell tracking; Magnetic resonance imaging (MRI); Ferritin; Superparamagnetic iron oxide (SPIO)> < 0.0001) and all other controls used. However, none of the techniques could be used to determine graft rejection in vivo, which is imperative for understanding cell behavior following transplantation. We conclude that in order for cell survival to be monitored in preclinical and clinical settings, another molecular imaging technique must be employed, including perhaps multimodal imaging, which would utilize MRI along with another imaging modality.

Key words: Neural progenitor cells; Cell tracking; Magnetic resonance imaging (MRI); Ferritin; Superparamagnetic iron oxide (SPIO)

Received June 26, 2014; final acceptance September 8, 2015. Online prepub date: July 8, 2015.
1Present address: University of Wisconsin-Madison, Department of Medicine, Madison, WI, USA.
Address correspondence to Clive N. Svendsen, 8700 Beverly Boulevard, AHSP A8404, Los Angeles, CA 90048, USA. Tel: +1-310-248-8072; Fax: +1-310-248-8066; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it



Cell Transplantation, Vol. 25, pp. 593-607, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689460
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Intrathecal Transplantation of Embryonic Stem Cell-Derived Spinal GABAergic Neural Precursor Cells Attenuates Neuropathic Pain in a Spinal Cord Injury Rat Model

Insik Hwang,*†1 Suk-Chan Hahm,†‡1 Kyung-Ah Choi,*§1 Sung-Ho Park,‡ Hyesun Jeong,*† Ji-Hye Yea,†‡ Junesun Kim,†‡ and Sunghoi Hong*†¶

*School of Biosystem and Biomedical Science, College of Health Science, Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea
†Department of Public Health Sciences, Graduate School, Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea
‡Department of Physical Therapy, College of Health Science, Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea
§Department of Chemistry, College of Science; Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea
¶Department of Integrated Biomedical and Life Science, Graduate School, Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea

Neuropathic pain following spinal cord injury (SCI) is a devastating disease characterized by spontaneous pain such as hyperalgesia and allodynia. In this study, we investigated the therapeutic potential of ESC-derived spinal GABAergic neurons to treat neuropathic pain in a SCI rat model. Mouse embryonic stem cell–derived neural precursor cells (mESC-NPCs) were cultured in media supplemented with sonic hedgehog (SHH) and retinoic acid (RA) and efficiently differentiated into GABAergic neurons. Interestingly, low doses of SHH and RA induced MGE-like progenitors, which expressed low levels of DARPP32 and Nkx2.1 and high levels of Irx3 and Pax6. These cells subsequently generated the majority of the DARPP32− GABAergic neurons after in vitro differentiation. The spinal mESC-NPCs wereintrathecally transplanted into the lesion area of the spinal cord around T10–T11 at 21 days after SCI. The engrafted spinal GABAergic neurons remarkably increased both the paw withdrawal threshold (PWT) below the level of the lesion and the vocalization threshold (VT) to the level of the lesion (T12, T11, and T10 vertebrae), which indicates attenuation of chronic neuropathic pain by the spinal GABAergic neurons. The transplanted cells were positive for GABA antibody staining in the injured region, and cells migrated to the injured spinal site and survived for more than 7 weeks in L4–L5. ThemESC-NPC-derived spinal GABAergic neurons dramatically attenuated the chronic neuropathic pain following SCI, suggesting that the spinal GABAergic mESC-NPCs cultured with low doses of SHH and RA could be alternative cell sources for treatment of SCI neuropathic pain by stem cell-based therapies.

Key words: Spinal cord injury (SCI); Neuropathic pain; Spinal GABAergic neurons; Intrathecal transplantation

Received February 14, 2015; final acceptance November 2, 2015. Online prepub date: September 16, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Sunghoi Hong, Ph.D., School of Biosystem and Biomedical Science, College of Health Science, Korea University, 145 Anam-dong, Sungbuk-gu, Seoul 136-701, Republic of Korea. Tel: +82-2-3290-5636; Fax: +82-2-916-5943; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Junesun Kim, P.T., Ph.D., Department of Physical Therapy, College of Health Science, Korea University, 145 Anam-dong, Sungbuk-gu, Seoul 136-701, Republic of Korea. Tel: +82-2-940-2834; Fax: +82-2-940-2830; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 609-614, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688939
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Brief Communication

Pancreatic Tissue Transplanted in TheraCyte™ Encapsulation Devices Is Protected and Prevents Hyperglycemia in a Mouse Model of Immune-Mediated Diabetes

Tobias Boettler,*†1 Darius Schneider,*1 Yang Cheng,* Kuniko Kadoya,‡ Eugene P. Brandon,‡ Laura Martinson,‡ and Matthias von Herrath*

*Type 1 Diabetes Center at the La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA
†University Hospital Freiburg, Department of Medicine II, Freiburg, Germany
ViaCyte, Inc., La Jolla, CA, USA

Type 1 diabetes (T1D) is characterized by destruction of glucose-responsive insulin-producing pancreatic β-cells and exhibits immune infiltration of pancreatic islets, where CD8 lymphocytes are most prominent. Curative transplantation of pancreatic islets is seriously hampered by the persistence of autoreactive immune cells that require high doses of immunosuppressive drugs. An elegant approach to confer graft protection while obviating the need for immunosuppression is the use of encapsulation devices that allow for the transfer of oxygen and nutrients, yet prevent immune cells from making direct contact with the islet grafts. Here we demonstrate that macroencapsulation devices (TheraCyte™) loaded with neonatal pancreatic tissue and transplanted into RIP-LCMV.GP mice prevented disease onset in a model of virus-induced diabetes mellitus. Histological analyses revealed that insulin-producing cells survived within the device in animal models of diabetes. Our results demonstrate that these encapsulation devices can protect from an immune-mediated attack and can contain a sufficient amount of insulin-producing cells to prevent overt hyperglycemia.

Key words: Type 1 diabetes (T1D); Encapsulation devices; Diabetes immunology; Virus-induced diabetes; RIP-LCMV

Received July 29, 2013; final acceptance August 25, 2015. Online prepub date: August 21, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Professor Matthias von Herrath, Type 1 Diabetes Center, La Jolla Institute for Allergy and Immunology, 9420 Athena Circle, La Jolla, CA 92037, USA. Tel: 858-752-6500; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 615, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X691006
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

ERRATUM

The following was originally published in Volume 20, Number 7, pages 1015–1031, 2011 (DOI: 10.3727/096368910X543402). The surname of one of the authors was not spelled correctly in the original article (Annalisa Gastadello). The correct spelling of this author’s name is Annalisa Gastaldello and is corrected in the information below.

Autologous Transplantation of Amniotic Fluid-Derived Mesenchymal Stem Cells Into Sheep Fetuses

S. W. Steven Shaw,*†‡ Sveva Bollini,† Khalil Abi Nader,* Annalisa Gastaldello,† Vedanta Mehta,* Elisa Filppi,* Mara Cananzi,† H. Bobby Gaspar,§ Waseem Qasim,§ Paolo De Coppi,†1 and Anna L. David*1

*Prenatal Cell and Gene Therapy Group, Institute for Women’s Health, University College London, London, UK
†Surgery Unit, Institute of Child Health, University College London, London, UK
‡Department of Obstetrics and Gynaecology, Chang Gung Memorial Hospital at Linkou and Chang Gung University, College of Medicine, Taoyuan, Taiwan
§Department of Paediatric Immunology, Institute of Child Health, University College London, London, UK

Long-term engraftment and phenotype correction has been difficult to achieve in humans after in utero stem cell transplantation mainly because of allogeneic rejection. Autologous cells could be obtained during gestation from the amniotic fluid with minimal risk for the fetus and the mother. Using a sheep model, we explored the possibility of using amniotic fluid mesenchymal stem cells (AFMSCs) for autologous in utero stem cell/gene therapy. We collected amniotic fluid (AF) under ultrasound-guided amniocentesis in early gestation pregnant sheep (n = 9, 58 days of gestation, term = 145 days). AFMSCs were isolated and expanded in all sampled fetal sheep. Those cells were transduced using an HIV vector encoding enhanced green fluorescent protein (GFP) with 63.2% (range 38.3–96.2%) transduction efficiency rate. After expansion, transduced AFMSCs were injected into the peritoneal cavity of each donor fetal sheep at 76 days under ultrasound guidance. One ewe miscarried twin fetuses after amniocentesis. Intraperitoneal injection was successful in the remaining 7 fetal sheep giving a 78% survival for the full procedure. Tissues were sampled at postmortem examination 2 weeks later. PCR analysis detected GFP-positive cells in fetal tissues including liver, heart, placenta, membrane, umbilical cord, adrenal gland, and muscle. GFP protein was detected in these tissues by Western blotting and further confirmed by cytofluorimetric and immunofluorescence analyses. This is the first demonstration of autologous stem cell transplantation in the fetus using AFMSCs. Autologous cells derived from AF showed widespread organ migration and could offer an alternative way to ameliorate prenatal congenital disease.

Key words: Autologous stem cell transplantation; Amniotic fluid stem cell; Sheep; In utero

Received February 22, 2010; final acceptance November 8, 2010. Online prepub date: November 19, 2010.
1These authors provided equal contribution to this work.
Address correspondence to Anna L. David, M.D., Ph.D., Institute for Women’s Health, University College London, 86-96 Chenies Mews, London, WC1E 6HX, UK. Tel: +44-20 7679-6651; Fax: +44-20 7383-7429; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Paolo de Coppi, M.D., Ph.D., Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1EH, UK. Tel: +44-20 7905 2641; Fax: +44-20 7404 618; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it