Cell Transplantation 24(6) Abstracts

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Cell Transplantation, Vol. 24, pp. 955-970, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681595
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Long-Term and Sustained Therapeutic Results of a Specific Promonocyte Cell Formulation in Refractory Angina: ReACT® (Refractory Angina Cell Therapy) Clinical Update and Cost-Effective Analysis

Nelson Americo Hossne, Jr.,* Eduardo Cruz,†‡ Enio Buffolo,* Anna Carolina Teixeira de Siqueira Mac Dowell Coimbra,†‡ Janaina Machado,† Regina Coeli dos Santos Goldenberg,§ Germana Regazzi,†‡ Silvia Azevedo,† Adriana Luckow Invitti,†‡ Joao Nelson Rodrigues Branco,* Jose Salvador Rodrigues de Oliveira,¶ Noedir Antonio Groppo Stolf,# Leslie W. Miller,** and Paul R. Sanberg††‡‡

*Cardiovascular Surgery Division, Surgery Department, Paulista School of Medicine, Federal University of Sao Paulo, Sao Paulo, SP, Brazil
Cryopraxis Criobiologia Ltda, Polo de Biotecnologia do Rio de Janeiro – Cidade Universitaria, Rio de Janeiro, RJ, Brazil
Cellpraxis Bioengeneering, Polo de Biotecnologia do Rio de Janeiro – Cidade Universitaria, Rio de Janeiro, RJ, Brazil
§Physiology Department, Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil
¶Hematology Division, Paulista School of Medicine, Federal University of Sao Paulo, Sao Paulo, SP, Brazil
#Heart Institute, College of Medicine, University of Sao Paulo, Sao Paulo, SP, Brazil
**Cardiology Department, University of South Florida, Tampa, FL, USA
††Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida Morsani College of Medicine, Tampa, FL, USA
‡‡Office of Research and Innovation, University of South Florida, Tampa, FL, USA

Mononuclear stem cells have been studied for their potential in myocardial ischemia. In our previous published article, ReACT®
phase I/II clinical trial, our results suggest that a certain cell population, promonocytes, directly correlated with the perceived angiogenesis in refractory angina patients. This study is ReACT’s clinical update, assessing long-term sustained efficacy. The ReACT phase IIA/B noncontrolled, open-label, clinical trial enrolled 14 patients with refractory angina and viable ischemic myocardium, without ventricular dysfunction, who were not suitable for myocardial revascularization. The procedure consisted of direct myocardial injection of a specific mononuclear cell formulation, with a certain percentage of promonocytes, in a single series of multiple injections (24–90; 0.2 ml each) into specific areas of the left ventricle. Primary endpoints were Canadian Cardiovascular Society Angina Classification (CCSAC) improvement at the 12-month follow-up and ischemic area reduction (scintigraphic analysis) at the 12-month follow-up, in correlation with ReACT’s formulation. A recovery index (for patients with more than 1 year follow-up) was created to evaluate CCSAC over time, until April 2011. Almost all patients presented progressive improvement in CCSAC beginning 3 months (p = 0.002) postprocedure, which was sustained at the 12-month follow-up (p = 0.002), as well as objective myocardium ischemic area reduction at 6 months (decrease of 15%, p < 0.024) and 12 months (decrease of 100%, p < 0.004) The recovery index (n = 10) showed that the patients were graded less than CCSAC 4 for 73.9 ± 24.2% over a median follow-up time of 46.8 months. After characterization, ReACT’s promonocyte concentration suggested a positive correlation with CCSAC improvement (r = −0.575, p = 0.082). Quality of life (SF-36 questionnaire) improved significantly in almost all domains. Cost-effectiveness analysis showed decrease in angina-related direct costs. Refractory angina patients presented a sustained long-term improvement in CCSAC and myocardium ischemic areas after the procedure. The long-term follow-up and strong improvement in quality of life reinforce effectiveness. Promonocytes may play a key role in myocardial neoangiogenesisReACT dramatically decreased direct costs.

Key words: Promonocyte; Angiogenesis; Refractory angina; Bone marrow mononuclear cells; Refractory angina cell therapy

Received January 17, 2012; final acceptance April 25, 2014. Online prepub date: May 6, 2014.
Address correspondence to Nelson Americo Hossne, Jr., Cardiovascular Surgery Division, Surgery Department, Paulista School of Medicine, Federal University of Sao Paulo, Botucatu St., 740, Sao Paulo, SP, Brazil 04023-900. Tel: +55-11-8166-5050; Fax: +55-11-5052-0386; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Paul R. Sanberg, Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida Morsani College of Medicine, 12901 Bruce B. Downs Blvd. MDC 78, Tampa, FL 33612, USA. Tel: +1-813-974-3154; Fax: +1-813-974-3078; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 971-983, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675746
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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The Structural Basis of Functional Improvement in Response to Human Umbilical Cord Blood Stem Cell Transplantation in Hearts With Postinfarct LV Remodeling

Yong Chen,*†1 Lei Ye,‡§¶#1 Jia Zhong,*† Xin Li,* Chen Yan,* Margaret P. Chandler,** Steve Calvin,†† Feng Xiao,‡‡ Mesfin Negia,‡‡ Walter C. Low,‡‡ Jianyi Zhang,‡§¶# and Xin Yu*†**§§

*Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, USA
†Case Center for Imaging Research, Case Western Reserve University, Cleveland, OH, USA
‡Department of Medicine, University of Minnesota Medical School, Minneapolis, MN, USA
§Department of Radiology, University of Minnesota Medical School, Minneapolis, MN, USA
¶Cardiovascular Division, University of Minnesota Medical School, Minneapolis, MN, USA
#Center for Magnetic Resonance Research, University of Minnesota Medical School, Minneapolis, MN, USA
**Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, OH, USA
††Department of Obstetrics and Gynecology, Abbott Northwestern Hospital, Minneapolis, MN, USA
‡‡Department of Neurosurgery, University of Minnesota Medical School, Minneapolis, MN, USA
§§Department of Radiology, Case Western Reserve University, Cleveland, OH, USA

Cellular therapy for myocardial repair has been one of the most intensely investigated interventional strategies for acute myocardial infarction. Although the therapeutic potential of stem cells has been demonstrated in various studies, the underlying mechanisms for such improvements are poorly understood. In the present study, we investigated the long-term effects of stem cell therapy on both myocardial fiber organization and regional contractile function using a rat model of postinfarct remodeling. Human nonhematopoietic umbilical cord blood stem cells (nh-UCBSCs) were administered via tail vein to rats 2 days after infarct surgery. Animals were maintained without immunosuppressive therapy. In vivo and ex vivo MR imaging was performed on infarct hearts 10 months after cell transplantation. Compared to the age-matched rats exposed to the identical surgery, both global and regional cardiac functions of the nh-UCBSC-treated hearts, such as ejection fraction, ventricular strain, and torsion, were significantly improved. More importantly, the treated hearts exhibited preserved fiber orientation and water diffusivities that were similar to those in sham-operated control hearts. These data provide the first evidence that nh-UCBSC treatment may prevent/delay untoward structural remodeling in postinfarct hearts, which supports the improved LV function observed in vivo in the absence of immunosuppression, suggesting a beneficial paracrine effect occurred with the cellular therapy.

Key words: Umbilical cord blood stem cells; Ischemic heart injury; Diffusion tensor MR imaging; Displacement encoding; Myocardial wall strain; Fiber architecture

Received March 29, 2012; final acceptance November 26, 2013. Online prepub date: December 10, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Xin Yu, Sc.D., Department of Biomedical Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Jianyi Zhang, M.D., Ph.D., Mayo Mail Code 508, University of Minnesota Health Science Center, 420 Delaware St. SE, Minneapolis, MN 55455, USA. E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 985-1002, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675151
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Epithelial-to-Mesenchymal Transition Enhances the Cardioprotective Capacity of Human Amniotic Epithelial Cells

Rajika Roy,* Marian Kukucka,† Daniel Messroghli,† Désirée Kunkel,* Andreja Brodarac,* Kristin Klose,* Sven Geißler,* Peter Moritz Becher,* Sung Keun Kang,‡ Yeong-Hoon Choi,§ and ChristofStamm*†

*Berlin Brandenburg Center for Regenerative Therapies, Berlin, Germany
Deutsches Herzzentrum Berlin, Berlin, Germany
‡Stem Cell Center, RNL Bio, Seoul, Korea
§University of Cologne, Cologne, Germany

The amniotic epithelium consists of cells exhibiting mature epithelial cell characteristics, but also varying degrees of stemness. We tested the hypothesis that induction of epithelial-to-mesenchymal transition (EMT) in amniotic epithelial cells (AECs) derived from human placenta enhances their capacity to support the ischemic myocardium. In response to incubation with transforming growth factor-β1 (TGF-β1) protein, AECs lost their cobblestone morphology and acquired a fibroblastoid shape, associated withdownregulation of E-cadherin, upregulation of N-cadherin, Akt phosphorylation, and intracellular periostin translocation. EMT–AECs displayed greatly enhanced mobility and secreted gelatinase activity compared with naive AECs. The surface presentation of CD105 and CD73 decreased, and RNA microarray analysis mirrored the loss of epithelial characteristics and transcriptional profile. Unmodified AECs and EMT–AECs were then injected intramyocardially in fully immunocompetent mice after permanent LAD ligation, and heart function was followed by MRI as well as 2D speckle tracking echocardiography after 4 weeks. EMT–AEC-treated infarct hearts displayed better global systolic function and improved longitudinal strain rate in the area of interest. Although no signals of human cells were detectable by histology, infarct size was smaller in EMT–AEC-treated hearts, associated with fewer TUNEL-positive cells and upregulation of periostin, while blood vessel density was increased in both ACE- and EMT–AEC-treated hearts. We conclude that EMT enhances the cardioprotective effects of human AECs.

Key words: Amniotic epithelial cells (AECs); Transforming growth factor-b1 (TGF-b1); Myocardial infarction; Cell therapy

Received February 21, 2013; final acceptance November 1, 2013. Online prepub date: November 20, 2013.
Address correspondence to Christof Stamm, M.D., Deutsches Herzzentrum Berlin, Augustenburger Platz 1, 13353 Berlin, Germany. Tel: +49 30 4593 2109; Fax: +49 30 4593 2100; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1003-1012, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X680109
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Intramuscular Transplantation of Pig Amniotic Fluid-Derived Progenitor Cells Has Therapeutic Potential in a Mouse Model of Myocardial Infarction

Shao-Yu Peng,*1 Chih-Jen Chou,*1 Po-Jen Cheng,† Tse-Yang Tseng,‡ Winston Teng-Kui Cheng,‡§ S. W. Steven Shaw,†¶# and Shinn-Chih Wu*

*Institute of Biotechnology, National Taiwan University, Taipei, Taiwan
†Department of Obstetrics and Gynaecology, Chang Gung Memorial Hospital at Linkou and Chang Gung University, College of Medicine, Taoyuan, Taiwan
‡Department of Animal Science and Technology, National Taiwan University, Taipei, Taiwan
§Department of Animal Science and Biotechnology, Tunghai University, Taichung, Taiwan
¶Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan
#Prenatal Cell and Gene Therapy Group, Institute for Women’s Health, University College London, London, UK

Acute myocardial infarction (MI) is a fatal event that causes a large number of deaths worldwide. MI results in pathological remodeling and decreased cardiac function, which could lead to heart failure and fatal arrhythmia. Cell therapy is a potential strategy to repair the damage through enhanced angiogenesis or by modulation of the inflammatory process via paracrine signaling. Amniotic fluid-derived progenitor cells (AFPCs) have been reported to differentiate into several lineages and can be used without ethical concerns or risk of teratoma formation. Since pigs are anatomically, physiologically, and genetically similar to humans, and pregnant pigs can be an abundant source of AFPCs, we used porcine AFPCs (pAFPCs) as our target cells. Intramyocardial injection of AFPCs has been shown to cure MI in animal models. However, intramuscular transplantation of cells has not been extensively investigated. In this study, we investigated the therapeutic potential of intramuscular injection of pAFPCs on acute MI. MI mice were divided into 1) PBS control, 2) medium cell dose (1 × 106
cells per leg; cell-M), and 3) high cell dose (4 × 106 cells per leg; cell-H) groups. Cells or PBS were directly injected into the hamstring muscle 20 min after MI surgery. Four weeks after MI surgery, the cell-M and cell-H groups exhibited significantly better ejection fraction, significantly greater wall thickness, smaller infarct scar sizes, and lower LV expansion index compared to the PBS group. Using in vivo imaging, we showed that the hamstring muscles from animals in the cell-M and cell-H groups had RFP-positive signals. In summary, intramuscular injection of porcine AFPCs reduced scar size, reduced pathological remodeling, and preserved heart function after MI.

Key words: Myocardial infarction (MI); Amniotic fluid-derived progenitor cells (AFPCs); Pig; Cell transplantation

Received December 4, 2013; final acceptance March 12, 2014. Online prepub date: March 24, 2014.
1These authors provided equal contribution to this work.
Address correspondence to S. W. Steven Shaw, Department of Obstetrics and Gynaecology, Chang Gung Memorial Hospital at Linkou and Chang Gung University, College of Medicine, 5 Fu-Shin Street, Taoyuan 333, Taiwan. Tel: +886-3-3281200, ext. 8251; Fax: +886-3-3288252; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Shinn-Chih Wu, Institute of Biotechnology, National Taiwan University, Room 108, 50 Lane 155, Section 3, Keelong Road, Taipei, 10764 Taiwan. Tel: +886-2-33664147; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1013-1029, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681036
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Modulation of Alloimmune Responses by Interleukin-10 Prevents Rejection of Implanted Allogeneic Smooth Muscle Cells and Restores PostinfarctionVentricular Function

Sanjiv Dhingra,* Jun Wu,* Shu-Hong Li,* Jian Guo,* Xi-Ping Huang,* Anton Mihic,* Jim Hu,† Richard D. Weisel,* and Ren-Ke Li*

*Division of Cardiovascular Surgery, Toronto General Research Institute, University Health Network and Department of Surgery, Division of Cardiac Surgery, University of Toronto, Toronto, Ontario, Canada
†Department of Laboratory Medicine and Pathobiology (Pediatrics), University of Toronto and Physiology and Experimental Medicine Program, Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada

Interleukin-10 (IL-10) gene transduction into allogeneic smooth muscle cells (SMCs) was evaluated to improve the long-term benefits of allogeneic cell transplantation into infarcted myocardium. Allogeneic cells, including SMCs, have been demonstrated to restore cardiac function and repair the infarcted myocardium, but late rejection of the transplanted cells by the host immune system may reverse the benefits of cell therapy. In a rat myocardial infarction model, three groups of rats were injected with either unmodified autologous, unmodified allogeneic, or allogeneic + IL-10 SMCs into the infarct region. Three weeks later, most of the allogeneic cells were rejected, whereas autologous cells were engrafted in the myocardium. IL-10 gene transduction of the allogeneic SMCs significantly improved the cell survival. To understand the mechanism of this improved survival, we evaluated the host immune responses against the SMCs. Allogeneic SMCs expressing IL-10 decreased leukocyte-mediated cytotoxicity incoculture, decreased the number of cytotoxic CD8+
T-cells, and increased the number of CD4+CD25+ regulatory T-cells in vitro and in vivo. Furthermore, IL-10 prevented the production of antidonor antibodies by the recipients against the allogeneic SMCs. Transplantation of unmodified autologous SMCs, but not unmodified allogeneic SMCs, significantly improved fractional shortening and left ventricular dimensions compared to the media-injected control group. However, IL-10 gene-enhanced allogeneic SMCs improved ventricular function, increased wall thickness, and decreased scar length in association with their enhanced survival. We conclude that IL-10 gene-enhanced cell therapy with allogeneic SMCs prevents detrimental alloimmune responses in the recipient, thereby increasing the survival of transplanted allogeneic SMCs and more effectively restoring cardiac function.

Key words: Cell therapy; Myocardial infarction; Cardiac regeneration; Immune rejection; Interleukin-10 (IL-10)

Received November 25, 2013; final acceptance April 9, 2014. Online prepub date: April 22, 2014.
Address correspondence to Ren-Ke Li, M.D., Ph.D., MaRS Centre, Toronto Medical Discovery Tower, Room 3-702, 101 College St., Toronto, ON, Canada, M5G 1L7. Tel: +1-416-581-7492; Fax: +1-416-581-7493; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1031-1042, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679200
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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AktmTOR Pathway Inhibits Apoptosis and Fibrosis in Doxorubicin-Induced Cardiotoxicity Following Embryonic Stem Cell Transplantation

Dinender K. Singla

Biomolecular Science Center, Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL, USA

Doxorubicin
(DOX) is an effective chemotherapeutic drug used for the treatment of a variety of malignancies. Unfortunately, time and dose-dependent DOX therapy induces cardiotoxicity and heart failure. We previously reported that transplanted embryonic stem (ES) cells and the conditioned medium (CM) can repair and regenerate injured myocardium in acute DOX-induced cardiomyopathy (DIC). However, the effectiveness of ES cell and CM therapeutics has not been challenged in the chronic DIC model. To this end, the long-term impact of ES cells and CM on apoptosis, fibrosis, cytoplasmic vacuolization, oxidative stress, and their associated mediators were examined. Four weeks post-DIC, ES cells and CM-transplanted hearts showed a significant decrease in cardiac apoptotic nuclei, which was consequent to modulation of signaling molecules in the Akt pathway including PTEN, Akt, and mTOR. Cytoplasmic vacuolization was reduced following treatment with ES cells and CM, as was cardiac fibrosis, which was attributable to downregulation of MMP-9 activity. Oxidative stress, as evidenced by DHE staining and lipid peroxide concentration, was significantly diminished, and preservation of the antioxidant defense system was observed following CM and ES cell transplantation. In conclusion, our data suggest that transplanted ES cells and CM have long-term potentiation to significantly mitigate various adverse pathological mechanisms present in the injured chronic DIC heart.

Key words: Doxorubicin (DOX); Embryonic stem (ES) cells; Conditioned media (CM); Akt

Received June 28, 2013; final acceptance January 29, 2014. Online prepub date: March 3, 2014.
Address correspondence to Dinender K. Singla, Ph.D. F.A.H.A., Biomolecular Science Center, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Room 224, Orlando, FL 32816, USA. Tel: +1-407-823-0953; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1043-1052, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679219
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Embryonic Stem Cells and Released Factors Stimulate c-kit+/FLK-1+ Progenitor Cells and Promote Neovascularization in Doxorubicin-Induced Cardiomyopathy

Dinender K. Singla and Latifa S. Abdelli

Biomolecular Science Center, Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL, USA

Vascular apoptosis plays a pivotal role in the development and progression of a myriad of cardiac dysfunctions, but has yet to be investigated in doxorubicin-induced cardiomyopathy (DIC). Additionally, the neovascularization potential and resulting functional consequences of embryonic stem (ES) cells and factors released from these cells in the chronic DIC myocardium remain largely unknown. To this end, we transplanted conditioned media (CM) and ES cells in the DIC-injured heart and evaluated their potential to inhibit vascular cell death, activate endogenous c-kit+
and FLK-1+ cells, enhance neovascularization, and augment left ventricular dysfunction. Data presented suggest transplanted CM and ES cells significantly blunt vascular cell apoptosis consequent to DIC. Quantitative immunohistochemistry data demonstrate significantly increased c-kit+ and FLK-1+ cells, as well as enhanced differentiated CD31+ cells in the CM and ES cell groups relative to DIC controls. Heart function, including fractional shortening and ejection fraction, assessed by transthoracic echocardiography, was significantly improved following CM and ES cell transplantation. In conclusion, our data suggest that transplantation of CM and ES cells inhibit vascular apoptosis, activate endogenous c-kit+ and FLK-1+ cells and differentiate them into endothelial cells, enhance neovascularization, and improve cardiac function in the DIC-injured myocardium.

Key words: Doxorubicin; Embryonic stem cells; Cardiomyopathy; Neovascularization

Received June 28, 2013; final acceptance January 29, 2014. Online prepub date: March 3, 2014.

Address correspondence to Dinender K. Singla, Ph.D. F.A.H.A., Biomolecular Science Center, College of Medicine, University of Central Florida, 4000 Central Florida Blvd, Room 224, Orlando, FL 32816, USA. Tel: +1-407-823-0953; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1053-1066, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681054
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Reversibly Immortalized Mouse Articular Chondrocytes Acquire Long-Term Proliferative Capability While Retaining Chondrogenic Phenotype

Joseph D. Lamplot,*1 Bo Liu,*†1 Liangjun Yin,*† Wenwen Zhang,*‡ Zhongliang Wang,*† Gaurav Luther,* Eric Wagner,* Ruidong Li,*† Guoxin Nan,*† Wei Shui,*† Zhengjian Yan,*† Richard Rames,* Fang Deng,*§ Hongmei Zhang,*† Zhan Liao,*¶ Wei Liu,*† Junhui Zhang,*† Zhonglin Zhang,*# Qian Zhang,*† Jixing Ye,*** Youlin Deng,*† Min Qiao,*† Rex C. Haydon,* Hue H. Luu,* Jovito Angeles,* Lewis L. Shi,* Tong-Chuan He,*† and Sherwin H. Ho*

*Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, IL, USA
†Department of Orthopaedic Surgery, the First Affiliated Hospital, and Ministry of Education Key Laboratory of Diagnostic Medicine and the Affiliated Hospitals of Chongqing Medical University, Chongqing, China
‡Department of Laboratory Medicine, the Affiliated Hospital, Binzhou Medical University, Yantai, Shandong, China
§Department of Cell Biology, Third Military Medical University, Chongqing, China
¶Department of Orthopaedic Surgery, Xiang-Ya Hospital of Central South University, Changsha, China
#Department of Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China
**School of Bioengineering, Chongqing University, Chongqing, China

Cartilage tissue engineering holds great promise for treating cartilaginous pathologies including degenerative disorders and traumatic injuries. Effective cartilage regeneration requires an optimal combination of biomaterial scaffolds, chondrogenic seed cells, and biofactors. Obtaining sufficient chondrocytes remains a major challenge due to the limited proliferative capability of primary chondrocytes. Here we investigate if reversibly immortalized mouse articular chondrocytes (iMACs) acquire long-term proliferative capability while retaining the chondrogenic phenotype. Primary mouse articular chondrocytes (MACs) can be efficiently immortalized with a retroviral vector-expressing SV40 large T antigen flanked with Cre/loxP sites. iMACs exhibit long-term proliferation in culture, although the immortalization phenotype can be reversed by Cre recombinase. iMACs express the chondrocyte markers Col2a1 and aggrecan and produce chondroid matrix in micromass culture. iMACs form subcutaneous cartilaginous masses in athymic mice. Histologic analysis and chondroid matrix staining demonstrate that iMACs can survive, proliferate, and produce chondroid matrix. The chondrogenic growth factor BMP2 promotes iMACs to produce more mature chondroid matrix resembling mature articular cartilage. Taken together, our results demonstrate that iMACs acquire long-term proliferative capability without losing the intrinsic chondrogenic features of MACs. Thus, iMACs provide a valuable cellular platform to optimize biomaterial scaffolds for cartilage regeneration, to identify biofactors that promote the proliferation and differentiation of chondrogenic progenitors, and to elucidate the molecular mechanisms underlying chondrogenesis.

Key words: Articular cartilage; Chondrocytes; Chondrogenesis; Tissue engineering; Immortalization; Chondrogenic progenitors; Osteoarthritis

Received August 7, 2013; final acceptance April 16, 2014. Online prepub date: May 2, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Tong-Chuan He, M.D., Ph.D., Molecular Oncology Laboratory, The University of Chicago Medical Center, 5841 South Maryland Avenue, MC 3079, Chicago, IL 60637, USA. Tel. +1 (773) 702-7169; Fax: +1 (773) 834-4598; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Sherwin Ho, M.D., Department of Orthopaedic Surgery, The University of Chicago Medical Center, 5841 South Maryland Avenue, MC 3079, Chicago, IL 60637, USA. Tel. +1 (773) 702-5978; Fax: +1 (773) 702-0554; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1067-1083, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681018
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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In Situ Recruitment of Human Bone Marrow-Derived Mesenchymal Stem Cells Using Chemokines for Articular Cartilage Regeneration

Min Sung Park,*† Yun Hee Kim,† Youngmee Jung,‡ Soo Hyun Kim,‡ Jong Chul Park,§ Dong Suk Yoon,*† Sung-Hwan Kim,† and Jin Woo Lee*†

*Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, South Korea
†Department of Orthopaedic Surgery, Yonsei University College of Medicine, Seoul, South Korea
‡Biomaterials Research Center, Korea Institute of Science and Technology, Seoul, South Korea
§Department of Medical Engineering, Yonsei University College of Medicine, Seoul, South Korea

Bone marrow-derived mesenchymal stem cells (BMSCs) are a good cell source for regeneration of cartilage as they can migrate directly to the site of cartilage injury and differentiate into articular chondrocytes. Articular cartilage defects do not heal completely due to the lack of chondrocytes or BMSCs at the site of injury. In this study, the chemotaxis of BMSCs toward chemokines, which may give rise to a complete regeneration of the articular cartilage, was investigated. CCR2, CCR4, CCR6, CXCR1, and CXCR2 were expressed in normal BMSCs and were increased significantly upon treatment with proinflammatory cytokines. BMSC migration was increased by MIP-3a and IL-8 more than by MCP-1 or SDF-1α. IL-8 and MIP-3α significantly enhanced the chemotaxis of BMSCs compared with MCP-1, SDF-1α, or PBS. Human BMSC recruitment to transplanted scaffolds containing either IL-8 or MIP-3α significantly increased in vivo compared to scaffolds containing PBS. Furthermore, IL-8- and MIP-3α-containing scaffolds enhanced tissue regeneration of an osteochondral defect site in beagle knee articular cartilage. Therefore, this study suggests that IL-8 and MIP-3α are the candidates that induce the regeneration of damaged articular cartilage.

Key words: Chemokines; IL-8; MIP-3α; Mesenchymal stem cells

Received December 13, 2011; final acceptance February 19, 2014. Online prepub date: April 22, 2014.
Address correspondence to Jin Woo Lee, M.D., Ph.D., Department of Orthopaedic Surgery-29, Yonsei University College of Medicine, Seoul 120-752, South Korea. Tel: +82-2-2228-2190; Fax: +82-2-363-1139; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1085-1096, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X680091
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Cell Therapy With G-CSF-Mobilized Stem Cells in a Rat Osteoarthritis Model

Ming-Wei Deng,* Shih-Jung Wei,† Tu-Lai Yew,‡ Po-Hui Lee,§ Tzu-Yu Yang,† Hen-Yi Chu,* and Shih-Chieh Hung†¶#**††

*Enhance Biomedical Ltd., Taipei, Taiwan
†Stem Cell Laboratory, Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan
‡Department of Biotechnology and Laboratory Science in Medicine, National Yang-Ming University, Taipei, Taiwan
§Institute of Oral Biology, National Yang-Ming University, Taipei, Taiwan
¶Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan
#Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
**Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan
††Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

G-CSF-mobilized peripheral blood stem cells (gm-PBSCs) offer a convenient cell source for treatment of hematopoietic and vascular disorders. Whether gm-PBSCs provide beneficial effects on skeleton diseases, such as osteoarthritis (OA), remains unknown. This study was undertaken to address the hypothesis that gm-PBSCs promote articular regeneration in OA. Here we studied the effect of single-dose intra-articular injection of gm-PBSCs from male donors delivered in hyaluronic acid (HA) on papain-induced OA in the knee joints of female Sprague–Dawley (SD) rats. Contralateral OA knee joints received single-dose HA alone and served as vehicle controls. We evaluated the histologic changes in glycosaminoglycan, type II collagen, type X collagen, modifiedMankin score, and cell apoptosis rate in the articular cartilage of rat knees. We demonstrated that gm-PBSCs were mobilized to the peripheral blood via G-CSF infusion for 5 days in SD rats with increasing CD34+
percentage up to 55-fold. We showed that gm-PBSCs inhibit progression of papain-induced OA via reducing articular surface irregularity, fibrillation, and erosion, preventing cellular necrosis and loss of chondrogenic proteins, such as glycosaminoglycan and type II collagen, at both 3 and 6 weeks after treatment. Moreover, gm-PBSCs reduced modified Mankin scores and cellular apoptosis rates compared with HA alone. Our findings demonstrate that HA plus gm-PBSCs, rather than HA alone, inhibits progression of OA in rats in vivo. Thus, intra-articular injection of gm-PBSCs is a convenient protocol for treating OA with consistent beneficial effects.

Key words: Granulocyte colony-stimulating factor; Osteoarthritis; Peripheral blood stem cells; Autologous; Preclinical trial

Received July 30, 2013; final acceptance March 4, 2014. Online prepub date: March 24, 2014.
Address correspondence to Shih-Chieh Hung, Department of Medical Research and Education, Taipei Veterans General Hospital, 201, Sec. 2, Shih-Pai Road, Taipei 11217, Taiwan. E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1097-1109, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679237
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Mesenchymal Stem Cells Increase Hippocampal Neurogenesis and Neuronal Differentiation by Enhancing the Wnt Signaling Pathway in an Alzheimer’s Disease Model

Se Hee Oh,*†1 Ha Na Kim,*†1 Hyun-Jung Park,*† Jin Young Shin,*† and Phil Hyu Lee*†

*Department of Neurology and Brain Research Institute, Yonsei University College of Medicine, Seoul, South Korea
†Severance Biomedical Science Institute, Yonsei University, Seoul, South Korea

Neurogenesis in the subgranular zone of the hippocampal dentate gyrus may act as an endogenous repair mechanism in Alzheimer’s disease (AD), and the Wnt signaling pathway has been suggested to closely modulate neurogenesis in amyloid-β (Aβ)-related AD models. The present study investigated whether mesenchymal stem cells (MSCs) would modulate hippocampal neurogenesis via modulation of the Wnt signaling pathway in a model of AD. In Aβ-treated neuronal progenitor cells (NPCs), the coculturewith MSCs increased significantly the expression of Ki-67, GFAP, SOX2, nestin, and HuD compared to Aβ treatment alone. In addition, MSC treatment in Aβ-treated NPCs enhanced the expression of β-catenin and Ngn1 compared to Aβ treatment alone. MSC treatment in Aβ-treated animals significantly increased the number of BrdU-ir cells in the hippocampus at 2 and 4 weeks compared to Aβ treatment alone. In addition, quantitative analysis showed that the number of BrdU and HuD double-positive cells in the dentate gyrus was significantly higher in the MSC-treated group than in controls or after Aβ treatment alone. These results demonstrate that MSC administration significantly augments hippocampal neurogenesis and enhances the differentiation of NPCs into mature neurons in AD models by augmenting the Wnt signaling pathway. The use of MSCs to modulate endogenous adult neurogenesis may have a significant impact on future strategies for AD treatment.

Key words: Mesenchymal stem cells (MSCs); Neurogenesis; Alzheimer’s disease (AD); Wnt signaling pathway

Received May 7, 2013; final acceptance February 4, 2014. Online prepub date: March 7, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Phil Hyu Lee, M.D., Ph.D., Department of Neurology, Yonsei University College of Medicine, 250 SeongsannoSeodaemun-gu, Seoul 120-752, South Korea. Tel: +82-2-2228-1608; Fax: +82-2-393-0705; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1111-1125, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675737
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparative Labeling of Equine and Ovine Multipotent Stromal Cells With Superparamagnetic Iron Oxide Particles for Magnetic Resonance Imaging In Vitro

Henriette Jülke,*† Christin Veit,* Iris Ribitsch,* Walter Brehm,‡ Eberhard Ludewig,§ and Uta Delling

*Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig, Germany
†FREY-TOX GmbH, Herzberg, Germany
‡Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, University of Leipzig, Leipzig, Germany
§Faculty of Veterinary Medicine, Department of Small Animal Medicine, University of Leipzig, Leipzig, Germany

The purpose of this study was to evaluate the use of three different superparamagnetic iron oxide (SPIO) particles for labeling of ovine and equine bone marrow (BM)-derived multipotent stromal cells (MSCs) in vitro. MSCs were obtained from five adult sheep and horses, respectively. After three passages (p3), cells were labeled with either 1) Molday ION Rhodamine B, 2) Endorem, 3) Resovist, or 4) remained unlabeled as control. Labeling efficiency, marker retention, and long-term detectability in MRI until p7 were evaluated. Further, proliferation capacity and trilineage differentiation as indicators for potential impact on stromal cell characteristics were assessed. MSCs of both species were successfully labeled with all three SPIO products. A high, exclusively intracellular, iron uptake was achieved by Molday ION Rhodamine B only. Labeling with Resovist led to prominent extracellular iron presence; labeling with Endorem was less efficient. During MRI, all labeled cells showed strong hypointense signals, contrary to unlabeled controls. Resovist induced the largest areas of hypointense signals, followed by Molday ION Rhodamine B and Endorem. MRI signal detectability decreased from p4 to p7. Proliferation, adipogenic, and osteogenic differentiation potential were not reduced by cell labeling compared to unlabeled cells. Chondrogenic differentiation capacity decreased with increasing amount of iron associated with the cells. Among the three products, Resovist and Molday were identified as promising labeling agents. While Resovist achieved superior results in most of the assessed parameters, Molday ION Rhodamine B ensured intracellular iron uptake without extracellular SPIO complexes and consistent hypointense signals on MRI.

Key words: Multipotent stromal cells (MSCs); Cell tracking; Superparamagnetic iron oxide (SPIO) particles; Magnetic resonance imaging

Received May 23, 2013; final acceptance November 21, 2013. Online prepub date: December 10, 2013.
Address correspondence to Henriette Jülke, Translational Centre for Regenerative Medicine, University of Leipzig, Philipp-Rosenthal-Str. 5, 04103 Leipzig, Germany. Tel: +49 341 9739610; Fax: +49 341 9739609; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1127-1138, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681702
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Efficient Engraftment of Human Induced Pluripotent Stem Cell-Derived Hepatocyte-Like Cells in uPA/SCID Mice by Overexpression of FNK, a Bcl-xLMutant Gene

Yasuhito Nagamoto,*† Kazuo Takayama,*†‡ Katsuhisa Tashiro,§ Chise Tateno,¶ Fuminori Sakurai,* Masashi Tachibana,* Kenji Kawabata,§ Kazuo Ikeda,# Yasuhito Tanaka,** and HiroyukiMizuguchi*†‡††

*Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan
†Laboratory of Hepatocyte Regulation, National Institute of Biomedical Innovation, Osaka, Japan
iPS Cell-based Research Project on Hepatic Toxicity and Metabolism, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan
§Laboratory of Stem Cell Regulation, National Institute of Biomedical Innovation, Osaka, Japan
PhoenixBio Co. Ltd., Hiroshima, Japan
#Department of Anatomy and Regenerative Biology, Graduate School of Medicine, Osaka City University, Osaka, Japan
**Department of Virology and Liver Unit, Graduate School of Medical Sciences, Nagoya City University, Nagoya, Japan
††The Center for Advanced Medical Engineering and Informatics, Osaka University, Osaka, Japan

Human liver chimeric mice are expected to be applied for drug toxicity tests and human hepatitis virus research. Human induced pluripotent stem cell-derived hepatocyte-like cells (iPSC-HLCs) are a highly attractive donor source for the generation of human liver chimeric mice because they can be produced on a large scale and established from an individual. Although these cells have been successfully used to generate human liver chimeric mice, there is still room for improvement in the repopulation efficiency. To enhance the repopulation efficacy, the human iPSC-HLCs were transduced with an adenovirus vector (Ad-FNK) expressing FNK, a hyperactive mutant gene from Bcl-xL, which was expected to inhibit apoptosis in the process of integration into liver parenchyma. We then transplanted Ad-FNK-transduced human iPSC-HLCs into urokinase-type plasminogen activator-transgenic severe combined immunodeficiency (uPA/SCID) mice (FNK mice) and evaluated the repopulation efficacy. The antiapoptoticeffects of the human iPSC-HLCs were enhanced by FNK overexpression in vitro. Human albumin levels in the transplanted mice were significantly increased by transplantation of Ad-FNK-transduced human iPSC-HLCs (about 24,000 ng/ml).Immunohistochemical analysis with an anti-human aAT antibody revealed greater repopulation efficacy in the livers of FNK mice than control mice. Interestingly, the expression levels of human hepatocyte-related genes in the human iPSC-HLCs of FNK mice were much higher than those in the human iPSC-HLCs before transplantation. We succeeded in improving the repopulation efficacy of human liver chimeric mice generated by transplanting the Ad-FNK-transduced human iPSC-HLCs into uPA/SCID mice. Our method using ectopic expression of FNK was useful for generating human chimeric mice with high chimerism.

Key words: Induced pluripotent stem cells (iPSCs); Hepatocyte; B-cell lymphoma extra large (Bcl-xL); FNK; Transplantation

Received November 17, 2013; final acceptance April 24, 2014. Online prepub date: May 6, 2014.
Address correspondence to Dr. Hiroyuki Mizuguchi, Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan. Tel: +81-6-6879-8185; Fax: +81-6-6879-8186; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1139-1153, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X680082
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Characterization of Liver-Specific Functions of Human Fetal Hepatocytes in Culture

Cinzia Maria Chinnici,* Francesca Timoneri,* Giandomenico Amico,* Giada Pietrosi,† Giovanni Vizzini,† Marco Spada,‡ Duilio Pagano,‡ Bruno Gridelli,*‡ and Pier Giulio Conaldi*

*Fondazione Ri.MED, Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, IRCCS-ISMETT, Palermo, Italy
Hepatology Unit, Department of Medicine, IRCCS-ISMETT, Palermo, Italy
‡Department of Surgery, IRCCS-ISMETT, Palermo, Italy

This study was designed to assess liver-specific functions of human fetal liver cells proposed as a potential source for hepatocyte transplantation. Fetal liver cells were isolated from livers of different gestational ages (16–22 weeks), and the functions of cell preparations were evaluated by establishing primary cultures. We observed that 20- to 22-week-gestation fetal liver cell cultures contained a predominance of cells with hepatocytic traits that did not divide in vitro but were functionally competent. Fetal hepatocytes performed liver-specific functions at levels comparable to those of their adult counterpart. Moreover, exposure to dexamethasone in combination with oncostatin M promptly induced further maturation of the cells through the acquisition of additional functions (i.e., ability to store glycogen and uptake of indocyanine green). In some cases, particularly in cultures obtained from fetuses of earlier gestational ages (16–18 weeks gestation), cells with mature hepatocytic traits proved to be sporadic, and the primary cultures were mainly populated by clusters of proliferating cells. Consequently, the values of liver-specific functions detected in these cultures were low. We observed that a low cell density culture system rapidly prompted loss of the maturehepatocytic phenotype with downregulations of all the liver-specific functions. We found that human fetal liver cells can be cryopreserved without significant loss of viability and function and evaluated up to 1 year in storage in liquid nitrogen. They might, therefore, be suitable for cell banking and allow for the transplantation of large numbers of cells, thus improving clinical outcomes. Overall, our results indicate that fetal hepatocytes could be used as a cell source for hepatocyte transplantation. Fetal liver cells have been used so far to treat end-stage liver disease. Additional studies are needed to include these cells in cell-based therapies aimed to treat liver failure and inborn errors of metabolism.

Key words: Human fetal hepatocytes; Primary cultures; Liver function; Cell transplant; Cryopreservation

Received June 7, 2013; final acceptance March 7, 2014. Online prepub date: March 24, 2014.
Address correspondence to Cinzia Maria Chinnici, Ph.D., Fondazione Ri.MED, Via E. Tricomi 5, 90127 Palermo, Italy. Tel: +39 091 21 92 496; Fax: +39 091 21 92 422; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1155-1165, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681027
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Preemptive Tolerogenic Delivery of Donor Antigens for Permanent Allogeneic Islet Graft Protection

Shusen Wang,*† Xiaomin Zhang,* Lei Zhang,‡ Jane Bryant,‡ Taba Kheradmand,‡ Bernhard J. Hering,§ Stephen D. Miller,¶ and Xunrong Luo*‡¶

*Comprehensive Transplant Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
†Organ Transplant Center and Key Laboratory for Critical Care Medicine of the Ministry of Health, Tianjin First Center Hospital, Tianjin, China
‡Division of Nephrology and Hypertension, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
§Schulze Diabetes Institute, Department of Surgery, University of Minnesota, Minneapolis, MN, USA
¶Department of Microbiology and Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

We have previously developed a robust regimen for tolerance induction in murine models of islet cell transplantation using pre- and posttransplant infusions of donor splenocytes (SPs) treated with a chemical cross-linker ethylcarbodiimide (ECDI). However, the requirement for large numbers of fresh donor SPs for ECDI coupling impairs its clinical feasibility, and additionally, the compatibility of this tolerance regimen with commonly used immunosuppressive drugs is largely unknown. In the current study, we demonstrate that equivalent tolerance efficacy for islet cell transplantation can be successfully achieved not only with a significantly lower dose of ECDI-SPs than originally established but also with culture-expanded donor B-cells or with soluble donor antigens in the form of donor cell lysate, which is ECDI coupled to recipient SPs. We further demonstrate that tolerance induced by donor ECDI-SPs is dependent on a favorable apoptotic-to-necrotic cell ratio post-ECDI coupling and is not affected by a transient course of conventional immunosuppressive drugs including tacrolimus and mycophenolate mofetil. While splenic antigen-presenting cells of the recipient play an important role in mediating the tolerogenic effects of donor ECDI-SPs, splenectomizedrecipients can be readily tolerized and appear to employ liver Kupffer cells for uptaking and processing of the ECDI-SPs. We conclude that infusion of donor ECDI-SPs is a versatile tolerance strategy that has a high potential for adaptation to clinically feasible regimens for tolerance trials for human islet cell transplantation.

Key words: Islet cell transplantation; Ethylene carbodiimide (ECDI); Tolerance; Apoptosis; Necrosis; Immunosuppressive drugs; Kupffer cells

Received November 24, 2013; final acceptance April 1, 2014. Online prepub date: April 22, 2014.
Address correspondence to Xunrong Luo, M.D., Ph.D., Division of Nephrology and Hypertension, Department of Medicine, Northwestern University FSM, 303 E. Chicago Ave., Tarry Building 4-751, Chicago, IL 60611, USA. Tel: +1-312-908-8147; Fax: +1-312-503-0622; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1167-1182, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679327
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Depletion of Alloreactive T-Cells by Anti-CD137–Saporin Immunotoxin

Sang C. Lee,*†1 Kwang W. Seo,*‡1 Hye J. Kim,* Sang W. Kang,§ Hye-Jeong Choi,¶ Ansuk Kim,# Byoung S. Kwon,** Hong R. Cho,*†† and Byungsuk Kwon*§

*Biomedical Research Center, Ulsan University Hospital, College of Medicine, University of Ulsan, Ulsan, Republic of Korea
†Personalized Medicine System R&D Center, Bio-support Co., Ltd., Anyang, Republic of Korea
‡Department of Internal Medicine, Ulsan University Hospital, College of Medicine, University of Ulsan, Ulsan, Republic of Korea
§School of Biological Sciences, University of Ulsan, Ulsan, Republic of Korea
¶Department of Pathology, Ulsan University Hospital, College of Medicine, University of Ulsan, Ulsan, Republic of Korea
#Department of Anesthesiology and Pain Medicine, Ulsan University Hospital, College of Medicine, University of Ulsan, Ulsan, Republic of Korea
**Division of Cell and Immunobiology and Research and Development Center for Cancer Therapeutics, National Cancer Center, Ulsan, Republic of Korea
††Department of Surgery, Ulsan University Hospital, College of Medicine, University of Ulsan, Ulsan, Republic of Korea

Depletion of alloreactive T-lymphocytes from allogeneic bone marrow transplants may prevent graft-versus-host disease (GVHD) without impairing donor cell engraftment, immunity, and the graft-versus-leukemia (GVL) effect. Alloreactive T-cells may be identified by their expression, upon activation, of CD137, a costimulatory receptor and putative surrogate marker for antigen-specific effector T-cells. In this context, we tested the use of anti-CD137–saporin immunotoxin to selectively deplete mouse and human alloreactive T-cells. Anti-CD137 antibodies were internalized by cells within 4 h of binding to the cell surface CD137, and anti-CD137–saporin immunotoxin effectively killed polyclonally activated T-cells or antigen-stimulated T-cells. Transfer of donor T-cells after allodepletion with anti-CD137–saporin immunotoxin failed to induce any evident expression of GVHD; however, a significant GVL effect was observed. Targeting of CD137 with an immunotoxin was also effective in killing polyclonallyactivated or alloreactive human T-cells. Our results indicate that anti-CD137–saporin immunotoxin may be used to deplete alloreactive T-cells prior to bone marrow transplantation and thereby prevent GVHD and the relapse of leukemia.

Key words: CD137; Graft-versus-host disease (GVHD); Graft-versus-leukemia (GVL) effect; Allodepletion; Immunotoxin

Received April 4, 2013; final acceptance February 7, 2014. Online prepub date: March 3, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Byungsuk Kwon, School of Biological Sciences, University of Ulsan, Ulsan, Korea. Tel: +82-52-259-1547; Fax: +82-52-259-2740; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Hong R. Cho, Department of Surgery, Ulsan University Hospital, Ulsan, Korea. Tel: +82-52-250-7100; Fax: +82-52-259-2740; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it