Cell Transplantation 21(7) Abstracts

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Cell Transplantation, Vol. 21, pp. 1341–1346, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636885
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved


Review
SUITO Index for Evaluation of Clinical Islet Transplantation

Morihito Takita*† and Shinichi Matsumoto*†

*Baylor Research Institute, Dallas, TX, USA
†The Institute of Medical Science, University of Tokyo, Tokyo, Japan

The major endpoints for clinical islet transplantation for type 1 diabetes are insulin independence and reduction of hypoglycemic episodes. Both endpoints are influenced by patients’ and physicians’ preferences regarding the use of exogenous insulin. Therefore, development of an objective endpoint for assessing clinical islet transplantation is desirable. HOMA-beta score is useful in assessing functional β-cell mass. However, this score uses blood insulin levels that are influenced by exogenous insulin injection and therefore is not suitable for patients who receive exogenous insulin. For assessing functional β-cell mass for type 1 diabetic patients after islet transplantation, we created the Secretory Unit of Islet Transplant Objects (SUITO) index using fasting C-peptide and fasting glucose. The formula of the SUITO index is fasting C-peptide (ng/ml)/[fasting blood glucose − 63 (mg/dl)] × 1500. We demonstrated that, within 1 month of islet transplantation, an average SUITO index of >26 was an excellent predictor of achieving insulin independence. In addition, daily SUITO index scores correlated with a reduction of insulin dose and adversely correlated with blood glucose levels during an intravenous glucose tolerance test. Other important endpoints, reduction of hypoglycemic episodes and quality of life, also correlated with the SUITO index. Thus, the SUITO index is excellent for assessing important endpoints (insulin independence, reduction of hypoglycemia, improved quality of life) after allogeneic islet transplantation.

Key words: SUITO index; Insulin independence; Hypoglycemic episodes; Islet transplantation

Received March 20, 2011; final acceptance July 23, 2011. Online prepub date: April 2, 2012.
Address correspondence to Morihito Takita, M.D., Ph.D., Baylor Research Institute, 1400 8th Ave., Fort Worth, TX 76104, USA. Tel: +1-817-922-2573; Fax: +1-817-922-4655; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1349–1360, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X623853
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved


Beneficial Effects of Ischemic Preconditioning on Pancreas Cold Preservation

Anthony R. Hogan,*† Marco Doni,*‡ R. Damaris Molano,* Melina M. Ribeiro,* Angela Szeto,* Lorenzo Cobianchi,*‡ Elsie Zahr-Akrawi,* Judith Molina,* Alessia Fornoni,*§¶ Armando J. Mendez,*§# Camillo Ricordi,*†§#**††‡‡ Ricardo L. Pastori,*§# and Antonello Pileggi*†**‡‡

*Diabetes Research Institute, University of Miami, Miami, FL, USA
†DeWitt-Daughtry Family Department of Surgery, University of Miami Leonard M. Miller School of Medicine, Miami, FL, USA
‡Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, University of Pavia, IRCCS Fondazione “San Matteo” Hospital, Pavia, Italy
§Department of Medicine, University of Miami Leonard M. Miller School of Medicine, Miami, FL, USA
¶Division of Nephrology, University of Miami Leonard M. Miller School of Medicine, Miami, FL, USA
#Division of Endocrinology, University of Miami Leonard M. Miller School of Medicine, Miami, FL, USA
**Department of Microbiology and Immunology, University of Miami Leonard M. Miller School of Medicine, Miami, FL, USA
††Jackson Memorial Hospital Transplant Institute, University of Miami, Miami, FL, USA
‡‡Department of Biomedical Engineering, University of Miami, Miami, FL, USA

Ischemic preconditioning (IPC) confers tissue resistance to subsequent ischemia in several organs. The protective effects are obtained by applying short periods of warm ischemia followed by reperfusion prior to extended ischemic insults to the organs. In the present study, we evaluated whether IPC can reduce pancreatic tissue injury following cold ischemic preservation. Rat pancreata were exposed to IPC (10 min of warm ischemia followed by 10 min of reperfusion) prior to _18 h of cold preservation before assessment of organ injury or islet isolation. Pancreas IPC improved islet yields (964 ± 336 vs. 711 ± 204 IEQ/pancreas; p = 0.004) and lowered islet loss after culture (33 ± 10% vs. 51 ± 14%; p = 0.0005). Islet potency in vivo was well preserved with diabetes reversal and improved glucose clearance. Pancreas IPC reduced levels of NADPH-dependent oxidase, a source of reactive oxygen species, in pancreas homogenates versus controls (78.4 ± 45.9 vs. 216.2 ± 53.8 RLU/μg; p = 0.002). Microarray genomic analysis of pancreata revealed upregulation of 81 genes and downregulation of 454 genes (greater than twofold change) when comparing IPC-treated glands to controls, respectively, and showing a decrease in markers of apoptosis and oxidative stress. Collectively, our study demonstrates beneficial effects of IPC of the pancreas prior to cold organ preservation and provides evidence of the key role of IPC-mediated modulation of oxidative stress pathways. The use of IPC of the pancreas may contribute to increasing the quality of donor pancreas for transplantation and to improving organ utilization.

Key words: Ischemic preconditioning; Pancreas; Cold ischemia; Cold preservation; Islets of Langerhans; Oxidative stress; NADPH oxidase; NADPH-dependent superoxide (NOX); Microarrays; Transcriptome; Rat

Received April 8, 2011; final acceptance July 20, 2011. Online prepub date: February 2, 2012.
Address correspondence to Antonello Pileggi, M.D., Ph.D., Associate Professor of Surgery, Microbiology & Immunology and Biomedical Engineering, Director, Preclinical Cell Processing & Translational Models Program, Cell Transplant Center-Diabetes Research Institute, University of Miami, 1450 NW 10th Avenue (R-134), Miami, FL 33136, USA. Tel: (305) 243-2924; Fax: (305) 243-4404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1361–1370, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637514
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Low Temperature Condition Prevents Hypoxia-Induced Islet Cell Damage and HMGB1 Release in a Mouse Model

Takeshi Itoh,* Koji Sugimoto,* Morihito Takita,* Masayuki Shimoda,† Daisuke Chujo,‡ Jeff A. SoRelle,§ Bashoo Naziruddin,¶ Marlon F. Levy,*¶ and Shinichi Matsumoto*

*Baylor Research Institute, Dallas, TX, USA
†Division of Cardiology, Department of Internal Medicine, Baylor University Medical Center, Baylor Heart and Vascular Institute, Dallas, TX, USA
‡Baylor Institute for Immunology Research, Dallas, TX, USA
§Institute of Biomedical Studies, Baylor University, Waco, TX, USA
¶Annette C. and Harold C. Simmons Transplant Institute, Dallas, TX, USA

One of the major issues in clinical islet transplantation is the poor efficacy of islet isolation. During pancreas preservation and islet isolation, islets suffer from hypoxia as islets are highly sensitive to hypoxic conditions. Cold preservation has been applied to minimize hypoxia-induced cell damage during organ preservation. However, the studies related to hypoxia-induced islet cell damage during islet isolation are limited. Recently, we demonstrated that mouse islets contain high levels of high-mobility group box 1 protein (HMGB1), and during proinflammatory cytokine-induced damage, islets release HMGB1 outside the cell. The released HMGB1 is involved in the initial events of early islet loss. In the present study, we hypothesize that low temperature conditions could prevent both hypoxia induced islet cell damage and HMGB1 release from islets in a mouse model. Isolated mouse islets underwent normoxic condition (95% air and 5% CO2) at 37°C or hypoxic conditions (1% O2, 5% CO2, and 94% N2) at 37°C (hypoxia-37°C islets), 22°C (hypoxia-22°C islets), or 4°C (hypoxia-4°C islets) for 12 h. In vitro and in vivo viability and functionality tests were performed. HMGB1, IL-6, G-CSF, KC, RANTES, MCP-1, and MIP-1α levels in the medium were measured. Low temperature conditions substantially reduced hypoxia-induced necrosis (p < 0.05) and apoptosis (p < 0.05). In addition, low temperature islet culture significantly increased the insulin secretion from islets by high glucose stimulation (p < 0.05). All of the recipient mice reversed diabetes after receiving the hypoxia-4°C islets but not after receipt of hypoxia-37°C or 22°C islets. The amounts of released HMGB1, IL-6, G-CSF, KC, RANTES, MCP-1, and MIP-1α were significantly reduced in the hypoxia-4°C islets compared to those of the hypoxia-37°C islets (p < 0.05). In conclusion, low temperature conditions could prevent hypoxia-induced islet cell damage, inflammatory reactions in islets, and HMGB1 release and expression. Low temperature conditions should improve the efficacy of isolated islets.

Key words: Islet transplantation; High-mobility group box 1 protein (HMGB1); Hypoxia; Preservation; Low temperature

Received November 15, 2010; final acceptance August 12, 2011. Online prepub date: March 27, 2012.
Address correspondence to Bashoo Naziruddin, Ph.D., Annette C. and Harold C. Simmons Transplant Institute, 3410 Worth St., Suite 950, Dallas, TX 75246, USA. Tel: 214-820-2662; Fax: 214-820-7142; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1371–1381, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640592
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Correlation of Released HMGB1 Levels With the Degree of Islet Damage in Mice and Humans and With the Outcomes of Islet Transplantation in Mice

Takeshi Itoh,* Morihito Takita,* Jeffrey A. SoRelle,† Masayuki Shimoda,‡ Koji Sugimoto,* Daisuke Chujo,§ Huanying Qin,¶ Bashoo Naziruddin,# Marlon F. Levy,# and Shinichi Matsumoto*

*Baylor Research Institute, Dallas, TX, USA
†Institute of Biomedical Studies, Baylor University, Waco, TX, USA
‡Division of Cardiology, Department of Internal Medicine, Baylor University Medical Center at Dallas and Baylor Heart and Vascular Institute, Dallas, TX, USA
§Baylor Institute for Immunology Research, Dallas, TX, USA
¶Institute for Health Care Research and Improvement, Baylor Health Care System, Dallas, TX, USA
#Annette C. and Harold C. Simmons Transplant Institute, Baylor University Medical Center at Dallas, Dallas, TX, USA

Establishing reliable islet potency assay is a critical and unmet issue for clinical islet transplantation. Recently, we reported that islets contained high levels of high mobility group box 1 (HMGB1) and damaged islets released HMGB1 in a mouse model. In this study, we hypothesized that the amount of released HMGB1 could reflect the degree of islet damage, and could predict the outcome of islet transplantation. Four groups of damaged mouse islets and three groups of damaged human islets were generated by hypoxic conditions. These islets were assessed by in vivo (transplantation) and in vitro (released HMGB1 levels, released C-peptide levels, PI staining, TUNEL staining, ATP/DNA, and glucose-stimulated insulin release test) assays. In addition, the ability of each assay to distinguish between noncured (n = 13) and cured (n = 7) mice was assessed. The curative rates of STZ-diabetic mice after receiving control, hypoxia-3h, hypoxia-6h, and hypoxia-24h mouse islets were 100%, 40%, 0%, and 0%, respectively. Only amounts of released HMGB1 and ratio of PI staining significant increased according to the degree of damages in both human and mouse islets. In terms of predictability of curing diabetic mice, amounts of released HMGB1 showed the best sensitivity (100%), specificity (100%), positive (100%), and negative predictive values (100%) among all the assays. The amount of released HMGB1 reflected the degree of islet damage and correlated with the outcome of islet transplantation in mice. Hence, released HMGB1 levels from islets should be a useful marker to evaluate the potency of isolated islets.

Key words: High mobility group box 1 (HMGB1); Islet; Transplantation; Potency assay; Hypoxia

Received April 26, 2011; final acceptance August 27, 2011. Online prepub date: April 25, 2012.
Address correspondence to Marlon F. Levy, M.D., F.A.C.S., Baylor All Saints Medical Center, 1400 8th Ave., Fort Worth, TX 76104, USA. Tel: 817-922-4649; Fax: 817-922-4655; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1383–1396, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640501
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Blockade of Endothelial Gi Protein Enhances Early Engraftment in Intraportal Cell Transplant to Mouse Liver

Javier Alfaro,* Montserrat Grau,† Manuel Serrano,* Ana I. Checa,‡ Luis Miguel Criado,§ Enrique Moreno,† Estela Paz-Artal,* Mario Mellado,‡ and Antonio Serrano*

*Cell Transplantation Unit, Department of Immunology, Hospital Universitario 12 de Octubre, Madrid, Spain
†Department of Surgery, Hospital Universitario 12 de Octubre, Madrid, Spain
‡ Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid, Spain
§Centro Nacional de Investigaciones Cardiovasculares, Madrid, Spain

The limited availability of liver donors and recent progress in cell therapy technologies has centered interest on cell transplantation as a therapeutic alternative to orthotopic liver transplant for restoring liver function. Following transplant by intraportal perfusion, the main obstacle to cell integration in the parenchyma is the endothelial barrier. Transplanted cells form emboli in the portal branches, inducing ischemia and reperfusion injury, which cause disruption of endothelial impermeability and activate the immune system. Approximately 95% of transplanted cells fail to implant and die within hours by anoikis or are destroyed by the host immune system. Intravascular perfusion of Bordetella pertussis toxin (PTx) blocks endothelial Gi proteins and acts as a reversible inducer of actin cytoskeleton reorganization, leading to interruption of cell confluence in vitro and increased vascular permeability in vivo. PTx treatment of the murine portal vascular tree 2 h before intraportal perfusion of embryonic stem cells facilitated rapid cell engraftment. By 2 h postperfusion, the number of implanted cells in treated mice was more than fivefold greater than in untreated controls, a difference that was maintained to at least 30 days posttransplant. We conclude that prior to cell transplant, PTx blockade of the Gi protein pathway in liver endothelium promotes rapid, efficient cell implantation in liver parenchyma, and blocks chemokine receptor signaling, an essential step in early activation of the immune system.

Key words: Hepatocyte transplant; Vascular endothelium; Drug effects

Received December 22, 2010; final acceptance August 5, 2011. Online prepub date: April 11, 2012.
Address correspondence to Antonio Serrano, Cell Transplantation Unit, Department of Immunology Hospital Universitario 12 de Octubre, 28041 Madrid, Spain. Tel: +34 91 390 8315; Fax: +34 390 8399; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1397–1406, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627589
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Clinical-Scale Isolation of Interleukin-2-Stimulated Liver Natural Killer Cells for Treatment of Liver Transplantation With Hepatocellular Carcinoma

Masahiro Ohira,* Seigo Nishida,* Panagiotis Tryphonopoulos,* Akin Tekin,* Gennaro Selvaggi,* Jang Moon,* David Levi,* Camillo Ricordi,† Kohei Ishiyama,‡ Yuka Tanaka,‡ Hideki Ohdan,‡ and Andreas G. Tzakis*

*Department of Surgery, Division of Liver and Gastrointestinal Transplantation, University of Miami Miller School of Medicine, Miami, FL, USA
†Cell Transplant Center, Diabetes Research Institute, University of Miami Miller School of Medicine, Miami, FL, USA
‡Department of Surgery, Division of Frontier Medical Science, Programs for Biomedical Research, Graduate School of Biomedical Science, Hiroshima University, Hiroshima, Japan

Tumor recurrence is the main limitation of liver transplantation (LT) in patients with hepatocellular carcinoma (HCC) and can be promoted by immunosuppressants. However, there is no prevention or treatment for HCC recurrence after LT. Here we describe a clinical-scale method for an adoptive immunotherapy approach that uses natural killer (NK) cells derived from deceased donor liver graft perfusate to prevent tumor recurrence after LT. Liver mononuclear cells (LMNCs) that were extracted from deceased donor liver graft perfusate contained a high percentage of NK cells (45.0 ± 4.0%) compared with peripheral blood mononuclear cells (PBMCs) (21.8 ± 5.2%) from the same donor. The CD69 activation marker and the natural cytotoxicity receptors, NKp44 and NKp46, were expressed at high levels in freshly isolated liver NK cells. Furthermore, interleukin-2 (IL-2)-stimulated NK cells showed greater upregulation of activation markers and the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), which is critical for NK cell-mediated antitumor cell death and increased production of interferon. Moreover, IL-2 stimulation induced LMNCs to exhibit a strong cytotoxicity against NK-susceptible K562 target cells compared with PBMCs (p < 0.01). Finally, we also showed that the final product contained a very low T-cell contamination (0.02 ± 106 cells/kg), which reduces the risk of graft-versus-host disease (GVHD). Collectively, our results suggest that the adoptive transfer of IL-2-stimulated NK cells from deceased donor liver graft perfusate could be a promising treatment for LT patients with HCC.

Key words: Natural killer cell; Immunotherapy; Innate immunity; Hepatocellular carcinoma; Current good manufacturing practice (cGMP)

Received December 15, 2010; final acceptance July 19, 2011. Online prepub date: March 16, 2012.
Address correspondence to Seigo Nishida, M.D., Ph.D., Department of Surgery, Division of Liver and Gastrointestinal Transplantation, University of Miami Miller School of Medicine, 1801 NW 9th Avenue, Miami, FL 33136, USA. Tel: 305-355-5760; Fax: 305-355-5793; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1407–1417, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X603648
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Human Adipose-Derived Mesenchymal Stromal Cells Injected Systemically Into GRMD Dogs Without Immunosuppression Are Able to Reach the Host Muscle and Express Human Dystrophin

N. M. Vieira, M. Valadares, E. Zucconi, M. Secco, C. R. Bueno Junior, V. Brandalise, A. Assoni, J. Gomes, V. Landini, T. Andrade, H. V. A. Caetano, M. Vainzof, and M. Zatz

Human Genome Research Center, Biosciences Institute, University of São Paulo, São Paulo, Brazil

Duchenne muscular dystrophy (DMD), a lethal X-linked disorder, is the most common and severe form of muscular dystrophies, affecting 1 in 3,500 male births. Mutations in the DMD gene lead to the absence of muscle dystrophin and a progressive degeneration of skeletal muscle. The possibility to treat DMD through cell therapy has been widely investigated. We have previously shown that human adipose-derived stromal cells (hASCs) injected systemically in SJL mice are able to reach and engraft in the host muscle, express human muscle proteins, and ameliorate the functional performance of injected animals without any immunosuppression. However, before starting clinical trials in humans many questions still need to be addressed in preclinical studies, in particular in larger animal models, when available. The best animal model to address these questions is the golden retriever muscular dystrophy (GRMD) dog that reproduces the full spectrum of human DMD. Affected animals carry a mutation that predicts a premature termination codon in exon 8 and a peptide that is 5% the size of normal dystrophin. These dogs present clinical signs within the first weeks and most of them do not survive beyond age two. Here we show the results of local and intravenous injections of hASCs into GRMD dogs, without immunosuppression. We observed that hASCs injected systemically into the dog cephalic vein are able to reach, engraft, and express human dystrophin in the host GRMD dystrophic muscle up to 6 months after transplantation. Most importantly, we demonstrated that injecting a huge quantity of human mesenchymal cells in a large-animal model, without immunosuppression, is a safe procedure, which may have important applications for future therapy in patients with different forms of muscular dystrophies.

Key words: Human multipotent mesenchymal stromal cells; Xenotransplantation; Muscular dystrophy; Therapy

Received November 30, 2010; final acceptance August 16, 2011. Online prepub date: October 14, 2011.
Address correspondence to Mayana Zatz, Human Genome Research Center, Institute of Biosciences, University of São Paulo, Rua do Matão, n.106-Cidade Universita´ria, Sa˜o Paulo-SP, Brasil-CEP: 05508-090. Tel: (55) (11) 3091-7581; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1419–1430, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627480
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Administration of a Soluble Activin Type IIB Receptor Promotes the Transplantation of Human Myoblasts in Dystrophic Mice

Raouia Fakhfakh,* Se-Jin Lee,† and Jacques P. Tremblay*

*Unité de recherche en Génétique Humaine, Centre de recherche de CHUL, CHUQ, Faculté de médecine, Université Laval, Québec, Canada
†Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD, USA

Duchenne muscular dystrophy (DMD) is a recessive disease caused by a dystrophin gene mutation. Myoblast transplantation permits the introduction of the dystrophin gene into dystrophic muscle fibers. However, this strategy has so far produced limited results. Modulation of transforming growth factor-β (TGF-β) superfamily signaling promotes skeletal muscle differentiation and growth and myogenic regeneration. We investigated the possibility that the combination of TGF-β superfamily signaling inhibition with myoblast transplantation might be an effective therapeutic approach in dystrophin-deficient patients. In vitro, blocking myostatin and other ligands with a soluble form of the extracellular domain of the activin IIB receptor (ActRIIB/Fc) upregulated the expression of myogenic differentiation factors and increased human myoblast fusion. In vivo, systemic inhibition of activin IIB receptor signaling by delivery of ActRIIB/Fc increased the success of the myoblast transplantation. This effect was further increased by forcing the mice to swim weekly to induce cycles of muscle degeneration and regeneration. Treatment of dystrophic mice with ActRIIB/Fc led to increased body weight, increased skeletal muscle mass, and improved myoblast transplantation. Thus, ActRIIB/Fc represents an effective therapeutic strategy for muscular dystrophies, and its effects are enhanced when combined with muscle exercise.

Key words: Myoblast transplantation; Duchenne muscular dystrophy (DMD); Transforming growth factor-β (TGF-β) superfamily; Activin receptor

Received February 10, 2011; final acceptance September 15, 2011. Online prepub date: March 22, 2012.

Address correspondence to Jacques P. Tremblay, Ph.D., Unité de recherche en Génétique Humaine, Centre de recherche du CHUL, 2705, Boulevard Laurier, Québec, G1v 4G2, Canada. Tel: (418) 654-2186; Fax: (418) 654-2207; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1431–1442, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640574
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Early Stage Foreign Body Reaction Against Biodegradable Polymer Scaffolds Affects Tissue Regeneration During the Autologous Transplantation of Tissue-Engineered Cartilage in the Canine Model

Yukiyo Asawa,* Tomoaki Sakamoto,* Makoto Komura,† Makoto Watanabe,* Satoru Nishizawa,* Yutaka Takazawa,‡ Tsuyoshi Takato,§ and Kazuto Hoshi*

*Departments of Cartilage & Bone Regeneration (Fujisoft), Tokyo University Graduate School of Medicine, Tokyo, Japan
†Department of Pediatric Surgery, Tokyo University Graduate School of Medicine, Tokyo, Japan
‡Department of Pathology, The University of Tokyo Hospital, Tokyo, Japan
§Departments of Sensory & Motor System Medicine, Tokyo University Graduate School of Medicine, Tokyo, Japan

To overcome the weak points of the present cartilage regenerative medicine, we applied a porous scaffold for the production of tissue-engineered cartilage with a greater firmness and a 3D structure. We combined the porous scaffolds with atelocollagen to retain the cells within the porous body. We conducted canine autologous chondrocyte transplants using biodegradable poly-L-lactic acid (PLLA) or poly-DL-lactic-co-glycolic acid (PLGA) polymer scaffolds, and morphologically and biochemically evaluated the time course changes of the transplants. The histological findings showed that the tissue-engineered constructs using PLLA contained abundant cartilage 1, 2, and 6 months after transplantation. However, the PLGA constructs did not possess cartilage and could not maintain their shapes. Biochemical measurement of the proteoglycan and type II collagen also supported the superiority of PLLA. The biodegradation of PLGA progressed much faster than that of PLLA, and the PLGA had almost disappeared by 2 months. The degraded products of PLGA may evoke a more severe tissue reaction at this early stage of transplantation than PLLA. The PLLA scaffolds were suitable for cartilage tissue engineering under immunocompetent conditions, because of the retarded degradation properties and the decrease in the severe tissue reactions during the early stage of transplantation.

Key words: Autologous chondrocyte transplantation; Canine; Tissue engineering; Biodegradable polymer

Received November 8, 2010; final acceptance September 10, 2011. Online prepub date: April 25, 2012.
Address correspondence to Kazuto Hoshi, M.D., Ph.D., Department of Cartilage & Bone Regeneration (Fujisoft), Graduate School of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8655, Japan. Tel: +81-3-3815-5411, ext. 37386; Fax: +81-3-5800-9891; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1443–1461, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653002
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Ex Vivo Akt/HO-1 Gene Therapy to Human Endothelial Progenitor Cells Enhances Myocardial Infarction Recovery

Keith R. Brunt,*† Jun Wu,† Zhilin Chen,* Daniel Poeckel,*‡ Ryan A. Dercho,§ Luis G. Melo,*¶1 Colin D. Funk,*‡ Christopher A. Ward,* and Ren-Ke Li†

*Department of Physiology, Queen’s University, Kingston, ON, Canada
†Department of Surgery, Division of Cardiovascular Surgery, University of Toronto and Toronto General Research Institute, University Health Network, Toronto, ON, Canada
‡Department of Biochemistry, Queen’s University, Kingston, ON, Canada
§Department of Pharmacology & Toxicology, Queen’s University, Kingston, ON, Canada
¶Department of Medicine, Queen’s University, Kingston, ON, Canada

The aim of this study was to evaluate the overexpression of genes central to cell survival and angiogenesis to enhance the function of human late outgrowth endothelial progenitor cells (EPCs) and their utility for infarct recovery. Ischemic myocardial injury creates a hostile microenvironment, which is characterized by hypoxia, oxidative stress, and inflammation. The infarct microenvironment prevents adhesion, survival, and integration of cell transplants that promote neovascularization. EPCs are dysfunctional as a result of risk factors in cardiovascular patients. Protein kinase B (Akt) and heme-oxygenase-1 (HO-1) are intracellular proteins that play an important role in angiogenesis and cell survival. Late outgrowth EPCs transduced ex vivo with Akt and HO-1 demonstrate improved adhesion to extracellular matrix, improved migration toward human cardiomyocytes, and an improved paracrine profile under stress. Enhanced late outgrowth EPCs reduce the tumor necrosis factor-α (TNF-α) burden both in vitro and in vivo, attenuating nuclear factor-κB (NF-κB) activity and promoting cell survival. Akt and HO-1 enhance late outgrowth EPC neovascularization, resulting in improved cardiac performance and reduced negative remodeling after myocardial infarction in nude mice. Alteration of the infarct microenvironment through gene modification of human late outgrowth EPCs enhances the function and integration of transplanted cells for restoration of cardiac function.

Key words: Vasculogenesis; Heart; Monocyte; Extracellular matrix; Growth factor; Cytokine; Secretome; Stem cell; Cardiac function; Regeneration

Received May 9, 2011; final acceptance September 5, 2011. Online prepub date: June 22, 2012.
1Sadly, this author is now deceased.
Address correspondence to Christopher A. Ward, Department of Physiology, Queen’s University, Kingston, ON, K7L 3N6, Canada. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Ren-Ke Li, MaRS Centre, Toronto Medical Discovery Tower, 101 College St., Room 3-702, Toronto, ON, M5G 1L7, Canada. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1463–1475, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640529
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Platelet-Mediated Mesenchymal Stem Cells Homing to the Lung Reduces Monocrotaline-Induced Rat Pulmonary Hypertension

Lei Jiang,* Xing Hui Song,† Pu Liu,* Chun Lai Zeng,‡ Zhang Sen Huang,* Lin Jing Zhu,* Yang Zi Jiang,† Hong Wei Ouyang,† and Hu Hu*

*Department of Pathology and Pathophysiology, School of Medicine, Zhejiang University, Hangzhou, China
†Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, Hangzhou, China
‡Department of Cardiology, Lishui Central Hospital, Lishui, China

Bone marrow mesenchymal stem cell (BM-MSC) transplantation has been suggested to be a promising method for the treatment of pulmonary arterial hypertension (PAH), a fatal disease currently without effective preventive/therapeutic strategies. However, the detailed mechanisms underlying BM-MSC therapy are largely unknown. We designed the present study to test the hypothesis that circulating platelets facilitate BM-MSC homing to the lung vasculature in a rat model of PAH induced by monocrotalin (MCT). A single subcutaneous administration of MCT induced a marked rise in right ventricular systolic pressure (RVSP) and the weight ratio of right to left ventricle plus septum (RV/LV+S) 3 weeks after injection. The injection of MSCs via tail vein 3 days after MCT significantly reduced the increase of RVSP and RV/LV+S. The fluorescence-labeled MSCs injected into the PAH rat circulation were found mostly distributed in the lungs, particularly on the pulmonary vascular wall, whereas cell homing was abolished by an anti-P-selectin antibody and the GPIIb/IIIa inhibitor tirofiban. Furthermore, using an in vitro flow chamber, we demonstrated that MSC adhesion to the major extracellular matrix collagen was facilitated by platelets and their P-selectin and GPIIb/IIIa. Therefore, the current study suggested that platelet-mediated MSC homing prevented the aggravation of MCT-induced rat PAH, via P-selectin and GPIIb/IIIa-mediated mechanisms.

Key words: Pulmonary arterial hypertension; Mesenchymal stem cells; Homing; Platelets

Received July 23, 2010; final acceptance August 14, 2011. Online prepub date: April 17, 2012.
Address correspondence to Hu Hu, Department of Pathology and Pathophysiology, School of Medicine, Zhejiang University, 388 Yu Hang Tang Road, 310058, Hangzhou, China. Tel: +86-571-882 085 17; Fax: +86-571-882 081 97; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Hong Wei Ouyang, Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, 388 Yu Hang Tang Road, 310058, Hangzhou, China. Tel: +86-571-882 082 62; Fax: +86-571-882 081 97; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1477–1492, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X601028
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Human Amnion Epithelial Cells Do Not Abrogate Pulmonary Fibrosis in Mice With Impaired Macrophage Function

Sean V. Murphy,* Suzane C. Shiyun,† Jean L. Tan,* Siow Chan,* Graham Jenkin,*† Euan M. Wallace,*† and Rebecca Lim*

*The Ritchie Centre, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia
†Department of Obstetrics and Gynecology, Monash Medical Centre, Monash University, Clayton, Victoria, Australia

Since current treatments for both acute and chronic lung diseases are less than ideal, there has been recent interest in the use of cell-based therapies for inflammatory lung disease. Specifically, human amnion epithelial cells (hAECs) have been shown to reduce bleomycin-induced lung injury and prevent subsequent loss of respiratory function, primarily through modulation of the host immune response. The precise mechanisms of this effect remain unclear. We aimed to investigate the potential of hAECs to mitigate bleomycin-induced lung injury in surfactant protein C deficient (Sftpc/) mice, which are highly susceptible to pulmonary injury as a result of impairment of macrophage function. Primary hAECs were administered to wild-type (Sftpc+/+) and Sftpc/ mice 24 h after exposure to bleomycin. Compared to Sftpc+/+ mice receiving bleomycin alone, Sftpc+/+ mice administered hAECs 24 h after bleomycin exposure had decreased expression of proinflammatory genes, decreased macrophage and neutrophil infiltration, fibrosis, collagen content, and α-smooth muscle actin as well as a significant improvement in lung function. Compared to Sftpc/ mice given bleomycin alone, Sftpc/ mice administered hAECs 24 h after bleomycin did not have a decrease in inflammatory gene expression or a reduction in macrophage pulmonary infiltration. Subsequently, Sftpc/ mice did not show any decrease in pulmonary fibrosis or improvement of lung function after hAEC administration. The ability of hAECs to mitigate bleomycin-induced lung injury is abolished in Sftpc/ mice, suggesting that hAECs require normal host macrophage function to exert their reparative effects.

Key words: Amnion epithelial cells; Macrophage; Bleomycin; Inflammation; Fibrosis

Received January 24, 2011; final acceptance August 5, 2011. Online prepub date: March 27, 2012.
Address correspondence to Prof. Euan Wallace, The Ritchie Centre, Monash Institute of Medical Research, Level 3, 27-31 Wright Street, Clayton, Victoria, 3168, Australia. Tel: 61-3-9594-5145; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1493–1502, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X647207
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Isolation and Characterization of Canine Wharton’s Jelly-Derived Mesenchymal Stem Cells

Min-Soo Seo,*†‡1 Sang-Bum Park,*†‡1 and Kyung-Sun Kang*†‡

*Adult Stem Cell Research Center, College of Veterinary Medicine, Seoul National University, Seoul, Korea
†Laboratory of Stem Cell and Tumor Biology, Department of Veterinary Public Health, College of Veterinary Medicine, Seoul National University, Seoul, Korea
‡BK 21 program for Veterinary Sciences, College of Veterinary Medicine, Seoul National University, Seoul, Korea

Wharton’s jelly is a known stem cell source in humans. Because stem cells might provide a potential therapeutic role in canines, many stem cell sources are studied for isolation and characterization in the canine system. So far, there have been no reports identifying canine Wharton’s jelly stem cells. In this study, we successfully isolated and characterized mesenchymal stem cells (MSCs) from canine Wharton’s jelly. Canine Wharton’s jelly-derived mesenchymal stem cells (cWJ-MSCs) that were grown in low-glucose DMEM medium have spindle-like shapes similar to human Wharton’s jelly stem cells. We characterized the immunophenotypes of canine Wharton’s jelly stem cells by FACS analysis and measured the cumulative population doubling level (CPDL). We investigated the differentiation of cWJ-MSCs with a trilineage differentiation assay to determine whether they were mesenchymal. Under various differentiation conditions, cWJ-MSCs presented chondrogenic, osteogenic, adipogenic, and neurogenic differentiation abilities in vitro. In conclusion, our results show that cWJ-MSCs might be a good source for stem cells. Furthermore, cWJ-MSCs might be useful as a cell therapy application for veterinary medicine.

Key words: Wharton’s jelly; Canine; Mesenchymal stem cells; Isolation

Received February 16, 2011; final acceptance September 18, 2011. Online prepub date: June 13, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Kyung-Sun Kang, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. Tel: +82-2-880-1241; Fax: +82-2-876-7610; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1503–1514, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640510
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Tumor Rejection Effects of Allorestricted Tumor Peptide-Specific CD4+ T Cells on Human Cervical Cancer Cell Xenograft in Nude Mice

Yinhong Song,1 Wei Sun,1 Xiufang Weng, Zhihui Liang, Qian Yu, Zhigang Wang, Lichen Ouyang, Jun Chen, Xiaolin Wu, Guanxin Shen, and Xiongwen Wu

Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

Generation of tumor specific alloreactive CD4+ T cells is important to circumvent tumor tolerance. Here, we generate allorestricted peptide-specific CD4+ T cells by coculture of lymphocytes and autologous monocytes bearing allogeneic HLA-DR15 molecule associated with its restricted peptide. Binding of a dimeric HLADR15/IgG1-Fc fusion protein (the dimer) to HLA-DR15 negative (HLA-DR15-ve) monocytes made the monocytes coated with the allogeneic epitope. An increased proliferation of CD4+ T cells and induction of Th1 cells appeared after coculturing of HLA-DR15-ve lymphocytes and the autologous monocytes loaded with the dimer. The cocultural bulks showed an increased frequency of the specific dimer-stained CD4+ T cells and the expanded CD4+ T cells exhibited an elevated IFN-γ production in response to specific TCR ligand. Tumor rejection effects of the allorestricted E7-specific CD4+ T cells raised by the coculture were observed in nude mice challenged with human cervical cancer cell SiHa expressing both HLA-DR15 and E7 antigens, as the tumor avoidance and life span of the mice were improved after adoptive transfer of the CD4+ T cells. This study may help to develop strategies to separate graft-versus-leukemia or graft-versus-tumor reaction from graft-versus-host disease, and add to the pool of human high-avidity TCRs specific for tumor or virus antigens.

Key words: Tumor rejection; Allorestricted; Tumor peptide specific; CD4+ T cell

Received February 19, 2011; final acceptance August 19, 2011. Online prepub date: April 17, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Xiongwen Wu, Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Tel: +86-27-83692611; Fax: +86-27-83693500; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1515–1527, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X647199
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Human Umbilical Cord Mesenchymal Stem Cells Support Nontumorigenic Expansion of Human Embryonic Stem Cells

Dah-Ching Ding,*† Woei-Cherng Shyu,‡§ Shinn-Zong Lin,‡§¶ Hwan-Wun Liu,†# Shih-Hwa Chiou,** and Tang-Yuan Chu*†

*Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, Tzu Chi University, Hualien, Taiwan
†Graduate Institute of Medical Science, Tzu Chi University, Hualien, Taiwan
‡Center for Neuropsychiatry, China Medical University and Hospital, Taichung, Taiwan
§Graduate Institute of Immunology, China Medical University, Taichung, Taiwan
¶China Medical University Beigang Hospital, Yunlin, Taiwan
#Department of Occupational Medicine, Buddhist Tzu-Chi Buddhist General Hospital, Hualien, Taiwan
**Institute of Clinical Medicine, Institute of Pharmacology, National Yang-Ming University & Taipei Veterans General Hospital, Taipei, Taiwan

The expansion of pluripotent human embryonic stem cells (hESCs) requires a culture on feeder layers of mouse embryonic fibroblasts (MEFs). The culture model often causes immunogenic contaminations such as xenocarbohydrate, and inevitably forms teratoma in vivo. This study tested human umbilical cord-derived mesenchymal stem cells (HUCMSCs) as the feeder for hESCs. Wharton’s jelly-derived HUCMSCs showed characteristics of MSCs and were easily maintained in a culture for over 20 passages. Under the mitomycin-inhibited HUCMSC feeder, hESCs maintained the features of embryonic stem cells (pluripotency and maintenance of normal karyotypes) after a prolonged culture of more than 20 passages. Notably, in extensive trials, no teratoma was formed in xenograft in NOD/SCID mice, but subsequent resumption of teratoma formation was noted upon transient coculturing with MEFs. Interestingly, among the four pluripotency-conferring genes, MYC and OCT4 were found to be downregulated in hESCs cocultured with HUCMSCs. Results of this study supported a nontumorigenic sustained culture of hESCs and did not form teratoma in vivo.

Key words: Embryonic stem cells; Umbilical cord; Mesenchymal stem cells; Tumorigenic

Received August 10, 2010; final acceptance September 1, 2011. Online prepub date: June 13, 2012.
Address correspondence to Tang-Yuan Chu, M.D., Ph.D., Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, Tzu Chi University, Hualien, Taiwan. Tel: 886-3-8561825; Fax: 886-3-8577161; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it

Cell Transplantation, Vol. 21, pp. 1529–1545, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640547
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Human Mesenchymal Stem Cells Exploit the Immune Response Mediating Chemokines to Impact the Phenotype of Glioblastoma

Helena Motaln,* Kristina Gruden,† Matjaž Hren,†‡ Christian Schichor,§ Monika Primon,* Ana Rotter,† and Tamara T. Lah*

*Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, Ljubljana, Slovenia
†Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia
‡Bioinstrumentation Laboratory, Centre of Excellence for Biosensors, Instrumentation and Process Control, Solkan, Slovenia
§Tumorbiological Laboratory, Neurosurgical Department, Ludwig-Maximilians-University, Munich, Germany

In contrast to the application of human mesenchymal stem cells (hMSCs) in regenerative medicine, only a limited number of studies are addressing their use in anticancer therapy. As the latter may represent a new hope to improve the survival of patients with glioblastoma multiformae (GBM), the most common and malignant form of the brain tumors, we aimed to investigate the interactions of hMSCs and GBM cells under in vitro conditions. Four hMSC clones and three different GBM cell lines were used to study their mutual paracrine interactions in cocultures compared to their monocultures, where cells were grown under the same experimental conditions. The effects on cell growth, proliferation, and invasion in Matrigel were quantified. Further, bioinformatics tools were used to relate these results to the data obtained from cytokine macroarrays and cDNA microarrays that revealed proteins and genes significantly involved in cellular crosstalk. We showed that hMSCs are responsible for the impairment of GBM cell invasion and growth, possibly via induction of their senescence. On the other hand, GBM cells inversely affected some of these characteristics in hMSCs. We found CCL2/MCP-1 to be the most significantly regulated chemokine during hMSC and U87-MG paracrine signaling in addition to several chemokines that may account for changed cocultured cells’ phenotype by affecting genes associated with proliferation (Pmepa-1, NF-κB, IL-6, IL-1b), invasion (EphB2, Sod2, Pcdh18, Col7A1, Gja1, Mmp1/2), and senescence (Kiaa1199, SerpinB2). As we functionally confirmed the role of CCL2/MCP-1 in GBM cell invasion we thereby propose a novel mechanism of CCL2/MCP-1 antimigratory effects on GBM cells, distinct from its immunomodulatory role. Significant alterations of GBM phenotype in the presence of hMSCs should encourage the studies on the naive hMSC use for GBM treatment.

Key words: Mesenchymal stem cells; Glioblastoma multiformae; CCL2/MCP-1

Received January 17, 2011; final acceptance September 15, 2011. Online prepub date: April 20, 2012.
Address correspondence to Helena Motaln, National Institute of Biology, Department of Genetic Toxicology and Cancer Biology, Večna pot 111, SI-1000 Ljubljana, Slovenia. Tel: +386 5 923 2870; Fax: +386 1 257 38 47; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1547–1559, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X600957
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Altered Oxygen Metabolism Associated to Neurogenesis of Induced Pluripotent Stem Cells Derived From a Schizophrenic Patient

Bruna da Silveira Paulsen,*1 Renata de Moraes Maciel,*1 Antonio Galina,† Mariana Souza da Silveira,‡ Cleide dos Santos Souza,* Hannah Drummond,* Ernesto Nascimento Pozzatto,* Hamilton Silva Junior,* Leonardo Chicaybam,§ Raffael Massuda,¶ Pedro Setti-Perdigão,# Martin Bonamino,§ Paulo Silva Belmonte-de-Abreu,¶ Newton Gonçalves Castro,# Helena Brentani,**††‡‡ and Stevens Kastrup Rehen*

*Laboratório Nacional de Células-Tronco Embrionárias, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
†Laboratório de Bioenergética e Fisiologia Mitocondrial, Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
‡Laboratório de Neurogênese, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
§Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro, Brazil
¶Departamento de Psiquiatria, Faculdade de Medicina, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil
#Laboratório de Farmacologia Molecular, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
**Departamento de Psiquiatria, Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil
††Instituto Nacional de Psiquiatria do Desenvolvimento, Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil
‡‡Laboratório de Investigação Médica, Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil

Schizophrenia has been defined as a neurodevelopmental disease that causes changes in the process of thoughts, perceptions, and emotions, usually leading to a mental deterioration and affective blunting. Studies have shown altered cell respiration and oxidative stress response in schizophrenia; however, most of the knowledge has been acquired from postmortem brain analyses or from nonneural cells. Here we describe that neural cells, derived from induced pluripotent stem cells generated from skin fibroblasts of a schizophrenic patient, presented a twofold increase in extramitochondrial oxygen consumption as well as elevated levels of reactive oxygen species (ROS), when compared to controls. This difference in ROS levels was reverted by the mood stabilizer valproic acid. Our model shows evidence that metabolic changes occurring during neurogenesis are associated with schizophrenia, contributing to a better understanding of the development of the disease and highlighting potential targets for treatment and drug screening.

Key words: Schizophrenia; Oxygen metabolism; Induced pluripotent stem cells; Reactive oxygen species (ROS) production

Received April 11, 2011; final acceptance August 28, 2011. Online prepub date: September 22, 2011.
1These authors provided equal contribution to this work.
Address correspondence to Stevens K. Rehen, National Laboratory for Embryonic Stem Cell Research, UFRJ, Hospital Universitário Clementino Fraga Filho-HUCFF, Rua Rodolpho Paulo Rocco, 255-4° andar-CEPEDIP-Cidade Universitária Rio de Janeiro-RJ, Brasil. Tel: +55 21 2562 2928; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1561–1575, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640484
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Bone Marrow Stromal Cell-Mediated Tissue Sparing Enhances Functional Repair After Spinal Cord Contusion in Adult Rats

Gaby J. Ritfeld,*† Rishi D. S. Nandoe Tewarie,‡ Katarina Vajn,* Sahar T. Rahiem,§ Andres Hurtado,§¶ Dane F. Wendell,§ Raymund A. C. Roos,† and Martin Oudega*#**

*Department of Physical Medicine & Rehabilitation, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
†Department of Neurology, Leiden University Medical Center, Leiden, The Netherlands
‡Department of Neurosurgery, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands
§International Center for Spinal Cord Injury, Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, MD, USA
¶Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA
#Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
**Department of Bioengineering, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

Bone marrow stromal cell (BMSC) transplantation has shown promise for repair of the spinal cord. We showed earlier that a BMSC transplant limits the loss of spinal nervous tissue after a contusive injury. Here, we addressed the premise that BMSC-mediated tissue sparing underlies functional recovery in adult rats after a contusion of the thoracic spinal cord. Our results reveal that after 2 months BMSCs had elicited a significant increase in spared tissue volumes and in blood vessel density in the contusion epicenter. A strong functional relationship existed between spared tissue volumes and blood vessel density. BMSC-transplanted rats exhibited significant improvements in motor, sensorimotor, and sensory functions, which were strongly correlated with spared tissue volumes. Retrograde tracing revealed that rats with BMSCs had twice as many descending brainstem neurons with an axon projecting beyond the contused spinal cord segment and these correlated strongly with the improved motor/sensorimotor functions but not sensory functions. Together, our data indicate that tissue sparing greatly contributes to BMSC-mediated functional repair after spinal cord contusion. The preservation/formation of blood vessels and sparing/regeneration of descending brainstem axons may be important mediators of the BMSC-mediated anatomical and functional improvements.

Key words: Transplantation; Bone marrow stromal cell (BMSC); Neuroprotection; Locomotion; Gridwalk; Allodynia

Received April 16, 2011; final acceptance September 7, 2011. Online prepub date: April 11, 2012.
Address correspondence to Martin Oudega, University of Pittsburgh School of Medicine, W1452 BSTWR, 200 Lothrop Street, Pittsburgh, PA 15213, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1577–1593, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636894
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Intracerebroventricular Transplantation of Human Bone Marrow-Derived Multipotent Progenitor Cells in an Immunodeficient Mouse Model of Mucopolysaccharidosis Type I (MPS-I)

Zhenhong Nan,* Laurie Shekels,† Oleg Ryabinin,* Carrie Evavold,* Matthew S. Nelson,† Shaukat A. Khan,† Robert J. Deans,‡ Robert W. Mays,‡ Walter C. Low,* and Pankaj Gupta†§

*Department of Neurosurgery, University of Minnesota, Minneapolis, MN, USA
†Hematology-Oncology Section, VA Medical Center, Minneapolis, MN, USA
‡Athersys Inc., Cleveland, OH, USA
§Hematology-Oncology-Transplantation Division, Department of Medicine, University of Minnesota, Minneapolis, MN, USA

Mucopolysaccharidosis type I (MPS-I; Hurler syndrome) is an inborn error of metabolism caused by lack of the functional lysosomal glycosaminoglycan (GAG)-degrading enzyme α-L-iduronidase (IDUA). Without treatment, the resulting GAG accumulation causes multisystem dysfunction and death within the first decade. Current treatments include allogeneic hematopoietic stem cell transplantation (HSCT) and enzyme replacement therapy. HSCT ameliorates clinical features and extends life but is not available to all patients, and inadequately corrects the most devastating features of the disease including mental retardation and skeletal deformities. Recent developments suggest that stem cells can be used to deliver needed enzymes to the central nervous system. To test this concept, we transplanted bone marrow-derived normal adult human MultiStem® cells into the cerebral lateral ventricles of immunodeficient MPS-I neonatal mice. Transplanted cells and human-specific DNA were detected in the hippocampal formation, striatum, and other areas of the central nervous system. Brain tissue assays revealed significant long-term decrease in GAG levels in the hippocampus and striatum. Sensorimotor testing 6 months after transplantation demonstrated significantly improved rotarod performance of transplanted mice in comparison to nontransplanted and sham-transplanted control animals. These results suggest that a single injection of MultiStem cells into the cerebral ventricles of neonatal MPS-I mice induces sustained reduction in GAG accumulation within the brain, and modest long-term improvement in sensorimotor function.

Key words: Mucopolysaccharidosis I (MPS-I); Stem cells; Glycosaminoglycans (GAGs); Proteoglycans; Transplantation

Received November 25, 2010; final acceptance August 12, 2011. Online prepub date: April 2, 2012.
Address correspondence to Pankaj Gupta, M.D., Professor of Medicine, University of Minnesota, Hematology/Oncology Section (111E), VA Medical Center, One Veterans Drive, Minneapolis, MN 55417, USA. Tel: (612)-467-4135; Fax: (612)-725-2149; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Walter C. Low, Ph.D., Professor, Department of Neurosurgery, University of Minnesota, 2001 Sixth Street SE, Minneapolis, MN 55455, USA. Tel: (612)-626-9203; Fax: (612)-626-9201; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1595–1602, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638900
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Brief Communication

Mild Cerebellar Neurodegeneration of Aged Heterozygous PCD Mice Increases Cell Fusion of Purkinje and Bone Marrow-Derived Cells

David Díaz,* Javier S. Recio,* Eduardo Weruaga,*1 and José R. Alonso*†1

*Laboratory of Neuronal Plasticity and Neurorepair, Institute for Neuroscience of Castile and León and Institute of Biomedical Research of Salamanca, Universidad de Salamanca, Salamanca, Spain
†Instituto de Alta Investigación, Universidad de Tarapacá, Arica, Chile

Bone marrow-derived cells have different plastic properties, especially regarding cell fusion, which increases with time and is prompted by tissue injury. Several recessive mutations, including Purkinje Cell Degeneration, affect the number of Purkinje cells in homozygosis; heterozygous young animals have an apparently normal phenotype but they undergo Purkinje cell loss as they age. Our findings demonstrate that heterozygous pcd mice undergo Purkinje cell loss at postnatal day 300, this slow but steadily progressing cell death starting sooner than has been reported previously and without massive reactive gliosis or inflammation. Here, transplantation of bone marrow stem cells was performed to assess the arrival of bone marrow-derived cells in the cerebellum in these heterozygous mice. Our results reveal that a higher number of cell fusion events occurs in heterozygous animals than in the controls, on days 150 and 300 postnatally. In sum, this study indicates that mild cell death promotes the fusion of bone marrow-derived cells with surviving Purkinje neurons. This phenomenon suggests new therapies for long-lasting neurodegenerative disorders.

Key words: Cell fusion; Cerebellum; Neurorepair; Purkinje cell degeneration; Purkinje cells; Transplant

Received February 16, 2011; final acceptance August 2, 2011. Online prepub date: March 27, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Dr. José R. Alonso, Institute for Neuroscience of Castilla y León, C/ Pintor Fernando Gallego 1E-37007, Salamanca, Spain. Tel: +34 923 294500, ext. 5325; Fax: +34 923 294750; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it