Cell Transplantation 22(7) Abstracts

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Cell Transplantation, Vol. 22, pp. 1101-1111, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653219
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Standardized Transportation of Human Islets: An Islet Cell Resource Center Study of More Than 2,000 Shipments

John S. Kaddis,* Matthew S. Hanson,† James Cravens,* Dajun Qian,* Barbara Olack,* Martha Antler,* Klearchos K. Papas,‡ Itzia Iglesias,§ Barbara Barbaro,¶ Luis Fernandez,† Alvin C. Powers,#** and Joyce C. Niland*

*Department of Information Sciences, City of Hope, Duarte, CA, USA
†Department of Surgery, University of Wisconsin, Madison, WI, USA
‡Department of Surgery, University of Minnesota, Minneapolis, MN, USA
§Department of Diabetes and Metabolic Diseases Research, City of Hope, Duarte, CA, USA
¶Division of Transplant Surgery, University of Illinois at Chicago, Chicago, IL, USA
#Division of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA
**VA Tennessee Valley Healthcare System, Nashville, TN, USA

Preservation of cell quality during shipment of human pancreatic islets for use in laboratory research is a crucial, but neglected, topic. Mammalian cells, including islets, have been shown to be adversely affected by temperature changes in vitro and in vivo, yet protocols that control for thermal fluctuations during cell transport are lacking. To evaluate an optimal method of shipping human islets, an initial assessment of transportation conditions was conducted using standardized materials and operating procedures in 48 shipments sent to a central location by eight pancreas-processing laboratories using a single commercial airline transporter. Optimization of preliminary conditions was conducted, and human islet quality was then evaluated in 2,338 shipments pre- and postimplementation of a finalized transportation container and standard operating procedures. The initial assessment revealed that the outside temperature ranged from a mean of -4.6 ± 10.3°C to 20.9 ± 4.8°C. Within-container temperature drops to or below 15°C occurred in 16 shipments (36%), while the temperature was found to be stabilized between 15°C and 29°C in 29 shipments (64%). Implementation of an optimized transportation container and operating procedure reduced the number of within-container temperature drops (£15°C) to 13% (n = 37 of 289 winter shipments), improved the number desirably maintained between 15°C and 29°C to 86% (n = 250), but also increased the number reaching or exceeding 29°C to 1% (n = 2; overall p < 0.0001). Additionally, postreceipt quality ratings of excellent to good improved pre- versus postimplantation of the standardized protocol, adjusting for preshipment purity/viability levels (p < 0.0001). Our results show that extreme temperature fluctuations during transport of human islets, occurring when using a commercial airline transporter for long distance shipping, can be controlled using standardized containers, materials, and operating procedures. This cost-effective and pragmatic standardized protocol for the transportation of human islets can potentially be adapted for use with other mammalian cell systems and is available online at http://iidp.coh.org/sops.aspx.

Key words: Human islets; Shipping protocol; Temperature control; Diabetes; Insulin

Received July 13, 2011; final acceptance June 4, 2012. Online prepub date: August 10, 2012.
Address correspondence to Joyce C. Niland, Ph.D., Chair, Department of Information Sciences, City of Hope National Medical Center, 1500 East Duarte Road, Duarte, CA 91010-3000, USA. Tel: +1-626-256-4673, ext. 63032; Fax: +1-626-301-8802; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1113-1121, 2013
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DOI: http://dx.doi.org/10.3727/096368912X657332
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Histidine-Tryptophan-Ketoglutarate and University of Wisconsin Solution Demonstrate Equal Effectiveness in the Preservation of Human Pancreata Intended for Islet Isolation: A Large-Scale, Single-Center Experience

Daniel H. Paushter, Meirigeng Qi, Kirstie K. Danielson, Tricia A. Harvat, Katie Kinzer, Barbara Barbaro, Sonny Patel, Sarah Z. Hassan, Jose Oberholzer, and Yong Wang

Department of Transplant/Surgery, University of Illinois at Chicago, Chicago, IL, USA

We previously reported a small-scale study on the efficacy of histidine-tryptophan-ketoglutarate (HTK) solution versus University of Wisconsin (UW) solution on pancreas preservation for islet isolation. In this large-scale, retrospective analysis (n = 252), we extend our initial description of the impact of HTK on islet isolation outcomes and include pancreatic digestion efficacy, purification outcomes, and islet size distribution. Multivariable linear regression analysis, adjusted for donor age, sex, BMI, cold ischemia time, and enzyme, demonstrated similar results for the HTK group (n = 95) and the UW group (n = 157), including postpurification islet yields (HTK: 289,702 IEQ vs. UW: 283,036 IEQ; p = 0.76), percentage of digested pancreatic tissue (HTK: 66.9% vs. UW: 64.1%; p = 0.18), and islet loss from postdigestion to postpurification (HTK: 24,972 IEQ vs. UW: 39,551 IEQ; p = 0.38). Changes in islet size between the postdigestion and postpurification stages were comparable within each islet size category for HTK and UW (p = 0.14–0.99). Tissue volume distribution across purification fractions and islet purity in the top fractions were similar between the groups; however, the HTK group had significantly higher islet purity in the middle fractions (p = 0.003–0.008). Islet viability and stimulation indices were also similar between the HTK and the UW groups. In addition, we analyzed a small sample of patients transplanted either with HTK (n = 7) or UW (n = 8) preserved islets and found no significant differences in posttransplant HbA1c, beta-score, and frequency of insulin independence. This study demonstrates that HTK and UW solutions offer comparable pancreas preservation for islet transplantation. More in vivo islet outcome data are needed for a complete analysis of the effects of HTK on islet transplantation.

Key words: Histidine-tryptophan-ketoglutarate (HTK); University of Wisconsin (UW); Organ preservation; Human pancreas; Islet isolation; Islet transplantation; Cellular edema

Received November 15, 2011; final acceptance July 15, 2012. Online prepub date: October 2, 2012.
Address correspondence to Yong Wang, Department of Transplant/Surgery, University of Illinois at Chicago, 840 South Wood Street, Clinical Science Building, Suite 502, Chicago, IL 60612, USA. Tel: +1-312-996-0851; Fax: +1-312-413-3483; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1123-1135, 2013
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DOI: http://dx.doi.org/10.3727/096368912X657440
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Macroporous Three-Dimensional PDMS Scaffolds for Extrahepatic Islet Transplantation

Eileen Pedraza,*†1 Ann-Christina Brady,*‡1 Christopher A. Fraker,*† R. Damaris Molano,* Steven Sukert,* Dora M. Berman,*‡ Norma S. Kenyon,*†‡§ Antonello Pileggi,*†‡§ Camillo Ricordi,*†‡¶ and Cherie L. Stabler*†‡

*Diabetes Research Institute, University of Miami, Miami, FL, USA
†Department of Biomedical Engineering, College of Engineering, University of Miami, Miami, FL, USA
‡DeWitt Daughtry Department of Surgery, Leonard M. Miller School of Medicine, University of Miami, Miami, FL, USA
§Department of Microbiology and Immunology, Leonard M. Miller School of Medicine, University of Miami, Miami, FL, USA
¶Department of Medicine, Leonard M. Miller School of Medicine, University of Miami, Miami, FL, USA

Clinical islet transplantation has demonstrated success in treating type 1 diabetes. A current limitation is the intrahepatic portal vein transplant site, which is prone to mechanical stress and inflammation. Transplantation of pancreatic islets into alternative sites is preferable, but challenging, as it may require a three-dimensional vehicle to confer mechanical protection and to confine islets to a well-defined, retrievable space where islet neovascularization can occur. We have fabricated biostable, macroporous scaffolds from poly(dimethylsiloxane) (PDMS) and investigated islet retention and distribution, metabolic function, and glucose-dependent insulin secretion within these scaffolds. Islets from multiple sources, including rodents, nonhuman primates, and humans, were tested in vitro. We observed high islet retention and distribution within PDMS scaffolds, with retention of small islets (<100 μm) improved through the postloading addition of fibrin gel. Islets loaded within PDMS scaffolds exhibited viability and function comparable to standard culture conditions when incubated under normal oxygen tensions, but displayed improved viability compared to standard two-dimensional culture controls under low oxygen tensions. In vivo efficacy of scaffolds to support islet grafts was evaluated after transplantation in the omental pouch of chemically induced diabetic syngeneic rats, which promptly achieved normoglycemia. Collectively, these results are promising in that they indicate the potential for transplanting islets into a clinically relevant, extrahepatic site that provides spatial distribution of islets as well as intradevice vascularization.

Key words: Islet transplantation; Scaffold; Extrahepatic sites; Omentum; Poly(dimethylsiloxane) (PDMS); Diabetes; Human islets; Rat islets; Nonhuman primate islets

Received October 4, 2011; final acceptance June 7, 2012. Online prepub date: October 2, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Cherie Stabler, Ph.D., 1450 NW 10th Avenue, Diabetes Research Institute, University of Miami, Miami, FL 33136, USA. Tel: +1-305-243-9768; Fax: +1-305-243-4404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1137-1146, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657486
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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The TheraCyteDevice Protects Against Islet Allograft Rejection in Immunized Hosts

Makiko Kumagai-Braesch,* Stella Jacobson,† Hiroki Mori,* Xiaohui Jia,* Tohru Takahashi,* Annika Wernerson,‡ Malin Flodström-Tullberg,† and Annika Tibell*

*Division of Transplantation Surgery, CLINTEC, Karolinska Institutet, Stockholm, Sweden
†Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden
‡Division of Renal Medicine, CLINTEC, Karolinska Institutet, Stockholm, Sweden

Clinically, many candidates for islet transplantation are already immunized, which increases their risk of graft rejection. Encapsulation of pancreatic islets using the TheraCyte™ device has been shown to protect against allograft rejection in nonimmunized recipients. However, the capacity of the TheraCyte™ device to prevent rejection in immunized recipients has not yet been studied. In this study, the protective capacity of the TheraCyte™ device was evaluated in an allogeneic rat model. Lewis rats were used as islet donors, and nonimmunized (control) and alloimmunized, diabetic Wistar–Furth (WF) rats were used as recipients. Graft survival was shorter in immunized recipients than in nonimmunized recipients (mean survival, 5.3 ± 2.7 and 9.3 ± 1.6 days, respectively, p < 0.01) when nonencapsulated islets were transplanted under the kidney capsule. When islets were transplanted into the TheraCyte™ device, graft function was maintained during the 6-month study period in both immunized and nonimmunized rats. In oral glucose tolerance tests performed at 1 month after transplantation, both groups had similar insulin and blood glucose levels indicating similar metabolic functions. Volume densities and absolute volumes of tissue inside the devices 6 months after transplantation were also comparable between the two groups, indicating that both groups maintained similar amounts of endocrine tissue. A higher number of IFN-γ-producing CD8+ T-cells were detected in immunized WF rats compared to control WF rats transplanted with encapsulated islets. This suggests that donor-specific alloreactivity in recipient rats was sustained throughout the study period. This study suggests that the TheraCyte™ device protects islet allografts also in immunized recipients. Our results further highlight the potential for using macroencapsulation to avoid immunosuppressive therapy in clinical islet transplantation.

Key words: Islet transplantation; Immunized recipients; TheraCyte™ device; Macroencapsulation; Alloantibodies; IFN-γ ELISpot

Received November 11, 2010; final acceptance June 3, 2012; Online prepub date: October 3, 2012.
Address correspondence to Makiko Kumagai-Braesch, Division of Transplantation Surgery, CLINTEC, Karolinska Institutet, Karolinska University Hospital Huddinge F82, 141 86, Stockholm, Sweden. Tel: +46 8 585 89520; Fax: +46 8 711 5202; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1147-1155, 2013
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DOI: http://dx.doi.org/10.3727/096368912X657233
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Administration of a Negative Vaccination Induces Hyporesponsiveness to Islet Allografts

M. M. Sklavos,*† G. M. Coudriet,*† M. Delmastro,*† S. Bertera,*† J. T. Coneybeer,*† J. He,*† M. Trucco,*† and J. D. Piganelli*†

*Division of Immunogenetics, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
†Rangos Research Center, Children’s Hospital of Pittsburgh, Pittsburgh, PA, USA

As a result of less than optimal outcomes the use of islet allografts as a standard insulin replacement therapy is limited to adults with a history of extreme glucose dysregulation and hypoglycemia unawareness. In this study, we examined the use of prophylactic immunotherapy to prevent islet allograft rejection in the absence of antirejection drugs. Our protocol to achieve allograft acceptance used a negative vaccination strategy that is comprised of apoptotic donor cells delivered in Incomplete Freund’s Adjuvant (IFA) 1 week prior to islet transplantation. The goal of this new protocol is to elicit hyporesponsiveness to alloantigen prior to islet transplantation. First, we examined our protocol without islet allograft transplants and determined that the negative vaccination was not globally immunosuppressive or immunostimulatory. Islet allograft experiments using fully MHC-mismatched islet donors and recipients demonstrated that the negative vaccination strategy induced long-term islet allograft acceptance. Upon rechallenge with alloantigen, the negative vaccination protocol successfully achieved hyporesponsiveness. In addition, the microenvironment at the site of the tolerant allograft revealed a decrease in proinflammatory mediators (IFN-γ, TNF-α) and an increase in the anti-inflammatory mediator IL-10, as well as increased expression of the master regulator of T-regulatory cells, FOXP3. Our data suggest that pretreating allograft recipients with apoptotic donor alloantigen delivered in IFA induced long-term islet allograft acceptance and glycemic control by introducing alloantigen to the recipient immune system in a nonimmunostimulatory manner prior to transplant.

Key words: Islet transplantation; Hyporesponsiveness; Negative vaccination; Allograft acceptance

Received February 2, 2011; final acceptance June 5, 2012. Online prepub date: October 1, 2012.

Address correspondence to Jon Piganelli, Ph.D., Children’s Hospital of Pittsburgh Rangos Research Center, 4401 Penn Avenue,

Pittsburgh, PA 15224, USA. Tel: +1 (412) 692-7498; Fax: +1 (412) 692-8131; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1157-1169, 2013
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DOI: http://dx.doi.org/10.3727/096368912X657350
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Inhibition of Recall Responses Through Complementary Therapies Targeting CD8+ T-Cell- and Alloantibody-Dependent Allocytotoxicity in Sensitized Transplant Recipients

Jason M. Zimmerer,* Phillip H. Horne,*† Lori A. Fiessinger,* Mason G. Fisher,* Kartika Jayashankar,* Sierra F. Garcia,* Mahmoud Abdel-Rasoul,‡ Nico van Rooijen,§ and Ginny L. Bumgardner*

*Department of Surgery, Comprehensive Transplant Center, The Ohio State University Medical Center, Columbus, OH, USA
†Integrated Biomedical Science Graduate Program, College of Medicine, The Ohio State University Medical Center, Columbus, OH, USA
‡Center for Biostatistics, The Ohio State University, Columbus, OH, USA
§Department of Molecular Cell Biology, Vrije University Medical Center, Amsterdam, The Netherlands

Allospecific T memory cell responses in transplant recipients arise from environmental exposure to previous transplantation or cross-reactive heterologous immunity. Unfortunately, these memory responses pose a significant barrier to the survival of transplanted tissue. We have previously reported that concurrent inhibition of CD154 and LFA-1 suppresses primary CD8-dependent rejection responses that are not controlled by conventional immunosuppressive strategies. We hypothesized that CD154- and LFA-1-mediated inhibition, by targeting activation as well as effector functions, may also be efficacious for the control of alloreactive CD8+ T-cell responses in sensitized hosts. We found that treatment with anti-LFA-1 mAb alone enhanced transplant survival and reduced CD8‑mediated cytotoxicity in sensitized CD4 KO recipients. However, treatment with anti‑CD154 mAb alone did not have an effect. Notably, when both CD4- and CD8-dependent rejection pathways are operative (wild-type sensitized recipients), LFA-1 significantly inhibited CD8-mediated in vivo allocytotoxicity but did not correspond with enhanced hepatocyte survival. We hypothesized that this was due to alloantibody‑mediated rejection. When anti-LFA-1 mAb treatment was combined with macrophage depletion, which we have previously reported impairs alloantibody‑mediated parenchymal cell damage, in vivo cytotoxic effector function was significantly decreased and was accompanied by significant enhancement of hepatocyte survival in sensitized wild-type recipients. Therefore, LFA-1 is a potent therapeutic target for reduction of CD8-mediated cytotoxicity in sensitized transplant recipients and can be combined with other treatments that target non‑CD8-mediated recall alloimmunity.

Key words: Alloantibody; CD154; CD8+ T-cells; Hepatocellular transplantation; LFA-1; Sensitized recipients

Received October 3, 2011; final acceptance July 10, 2012. Online prepub date: October 11, 2012.
Address correspondence to Ginny L. Bumgardner, M.D., Ph.D., F.A.C.S., The Ohio State University Medical Center, Department of Surgery, Division of Transplant, 395 W. 12th Avenue, 166 Doan Tower, Columbus, OH 43210-1250, USA. Tel: +1-614-293-6177; Fax: +1-614-293-4541; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1171-1183, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657431
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Effects of MSC Coadministration and Route of Delivery on Cord Blood Hematopoietic Stem Cell Engraftment

S. Carrancio,*†‡ C. Romo,*†‡ T. Ramos,*† N. Lopez-Holgado,*† S. Muntion,*† H. J. Prins,§ A. C. Martens,§ J. G. Briñón,¶ J. F. San Miguel,*†‡ M. C. del Cañizo,*†‡ and F. Sanchez-Guijo*†‡

*Servicio de Hematología, IBSAL-Hospital Universitario de Salamanca, Salamanca, Spain
†Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León and Red Nacional de Terapia Celular (Tercel, ISCIII), Salamanca, Spain
‡Centro de Investigación del Cáncer-IBMCC (Universidad de Salamanca-CSIC), Salamanca, Spain
§Department of Immunology and Department of Cell Biology, University Medical Center Utrecht, Utrecht, The Netherlands
¶Departamento de Biologia Celular y Patologia, Universidad de Salamanca, Salamanca, Spain

Hematopoietic stem cell transplantation (HSCT) using umbilical cord blood (UCB) progenitors is increasingly being used. One of the problems that may arise after UCB transplantation is an impaired engraftment. Either intrabone (IB) injection of hematopoietic progenitors or mesenchymal stem cell (MSC) coadministration has been proposed among the strategies to improve engraftment. In the current study, we have assessed the effects of both approaches. Thus, NOD/SCID recipients were transplanted with human UCB CD34+ cells administered either intravenously (IV) or IB, receiving or not bone marrow (BM)-derived MSCs also IV or IB (in the right femur). Human HSC engraftment was measured 3 and 6 weeks after transplantation. Injected MSCs were tracked weekly by bioluminescence. Also, lodgment within the BM niche was assessed at the latter time point by immunofluorescence. Our study shows regarding HSC engraftment that the number of BM human CD45+ cells detected 3 weeks after transplantation was significantly higher in mice cotransplanted with human MSCs. Moreover, these mice had a higher myeloid (CD13+) engraftment and a faster B-cell (CD19+) chimerism. At the late time point evaluated (6 weeks), human engraftment was higher in the group in which both strategies were employed (IB injection of HSC and MSC coadministration). When assessing human MSC administration route, we were able to track MSCs only in the injected femurs, whereas they lost their signal in the contralateral bones. These human MSCs were mainly located around blood vessels in the subendosteal region. In summary, our study shows that MSC coadministration can enhance HSC engraftment in our xenogenic transplantation model, as well as IB administration of the CD34+ cells does. The combination of both strategies seems to be synergistic. Interestingly, MSCs were detected only where they were IB injected contributing to the vascular niche.

Key words: Mesenchymal stem cells (MSCs); Cordon blood transplantation; Bone marrow microenvironment; Hematopoietic engraftment

Received October 11, 2011; final acceptance June 13, 2012. Online prepub date: October 2, 2012.
Address correspondence to Prof. F. Sánchez-Guijo, Servicio de Hematología, Hospital Universitario de Salamanca, Paseo de San Vicente 58-182, 37007 Salamanca, Spain. Tel: +34923291384; Fax: +34923294624; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1185-1199, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657288
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Proteomic Profiling of Secreted Proteins for the Hematopoietic Support of Interleukin-Stimulated Human Umbilical Vein Endothelial Cells

Gürkan Bal,* Julian Kamhieh-Milz,* Viktor Sterzer,* Muhammad Al-Samman,* Janusz Debski,† Oliver Klein,‡ Sundrela Kamhieh-Milz,* Sucharit Bhakdi,§ and Abdulgabar Salama*

*Institute for Transfusion Medicine, Charité Universitätsmedizin Berlin, Berlin, Germany
†Institute of Biochemistry and Biophysics, Polish Academy of Science, Warsaw, Poland
‡Berlin-Brandenburg Center for Regenerative Therapies, Charité Universitätsmedizin Berlin, Berlin, Germany
§Institute of Medical Microbiology and Hygiene, Johannes Gutenberg-University Mainz, Mainz, Germany

Human umbilical cord vein endothelial cells (HUVECs) secrete a number of factors that greatly impact the proliferation and differentiation of hematopoietic stem and progenitor cells (HSPCs). These factors remain largely unknown. Here, we report on the most comprehensive proteomic profiling of the HUVEC secretome and identified 827 different secreted proteins. Two hundred and thirty-one proteins were found in all conditions, whereas 369 proteins were identified only under proinflammatory conditions following IL-1β, IL-3, and IL-6 stimulation. Thirteen proteins including complement factor b (CFb) were identified only under IL-1β and IL-3 conditions and may potentially represent HSPC proliferation factors. The combination of bioinformatics and gene ontology annotations indicates the role of the complement system and its activation. Furthermore, CFb was found to be transcriptionally strongly upregulated. Addition of complement component 5b-9 (C5b-9) monoclonal antibody to the stem cell expansion assay was capable of significantly reducing their proliferation. This study suggests a complement-mediated cross-talk between endothelial cells and HSPCs under proinflammatory conditions.

Key words: Endothelial cells (ECs); Hematopoietic stem and progenitor cells (HSPCs); Expansion; Proteomics; Interleukin stimulated

Received August 5, 2011; final acceptance May 26, 2012. Online prepub date: October 1, 2012.
Address correspondence to Dr. Gürkan Bal, Institute for Transfusion Medicine, Campus Virchow-Klinikum, Charité Universitaetsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, Germany. Tel: +49-30-450 553139; Fax: +49-30-450 565904; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1201-1211, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657378
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Therapeutic Superiority for Cartilage Repair by CD271-Positive Marrow Stromal Cell Transplantation

Yutaka Mifune,*† Tomoyuki Matsumoto,*† Satoshi Murasawa,* Atsuhiko Kawamoto,* Ryosuke Kuroda,† Taro Shoji,*† Tomoya Kuroda,*† Tomoaki Fukui,*† Yohei Kawakami,*† Masahiro Kurosaka,† and Takayuki Asahara*‡

*Stem Cell Translational Research, Kobe Institute of Biomedical Research and Innovation, Kobe, Japan
†Department of Orthopedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan
‡Department of Regenerative Medicine Science, Tokai University School of Medicine, Kanagawa, Japan

Recent reports indicated that human isolated CD271+ bone marrow mesenchymal stromal cells (BM-MSCs) have a greater expansion and potential for multipotent differentiation including chondrogenesis than classical plastic adherent (PA) BM-MSCs in vitro. Therefore, we set up a hypothesis that CD271+ MSCs may have a greater chondrogenic potential than PA-MSCs in vitro and in vivo. We investigated the superiority of CD271+ MSCs on chondrogenesis using in vitro expansion and pellet culture system and in vivo rat model of cartilage defect when compared to PA-MSCs. In the in vitro study, CD271+ MSCs showed higher expansion potential and produced larger pellets with higher expressions of chondrogenic genes when compared to the control groups. During the culture, CD271 expression decreased, which resulted in decreased chondrogenesis. In the in vivo study, immunohistochemical staining demonstrated differentiated human chondrocytes identified as double-stained cells with human-specific collagen type 2 and human leukocyte antigen-ABC in CD271+ and PA groups. The number of double-stained cells was significantly higher in the CD271+ group than PA group. Real-time RT-PCR analysis of tissue RNA isolated from the chondral defect site for human-specific chondrogenic markers demonstrated a significantly higher expression in CD271+ group than PA group. Macroscopic examination of chondral defect sites at week 8 revealed glossy white and well-integrated repaired tissues in the CD271+ and PA groups, but not in the PBS group. The average histological score in the CD271+ group was significantly greater than in the other groups. Apoptosis analysis at the cell transplanted site with TUNEL staining showed that the CD271+ group had significantly fewer apoptotic chondrocytes compared with the PA group. These results indicate that CD271+ MSCs have a greater chondrogenic potential than PA-MSCs in both in vitro and in vivo conditions.

Key words: CD271; Mesenchymal stem cell (MSC); Cartilage repair; Bone marrow (BM)

Received September 3, 2009; final acceptance June 2, 2012. Online prepub date: October 3, 2012.
Address correspondence to Takayuki Asahara, M.D., Ph.D. or Tomoyuki Matsumoto, M.D., Ph.D., Stem Cell Translational Research, Kobe Institute of Biomedical Research and Innovation/RIKEN Center for Developmental Biology, 2-2 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan. Tel: +81-78-304-5772; Fax: +81-78-304-5263; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1213-1226, 2013
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DOI: http://dx.doi.org/10.3727/096368912X657224
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Low Molecular Weight Dextran Sulfate Binds to Human Myoblasts and Improves Their Survival After Transplantation in Mice

Thomas Laumonier,* Amandine Pradier,† Pierre Hoffmeyer,* Vincent Kindler,†1 and Jacques Menetrey*1

*Orthopaedic Surgery Service, Geneva University Hospitals and Faculty of Medicine, Geneva, Switzerland
†Hematology Service, Geneva University Hospitals and Faculty of Medicine, Geneva, Switzerland

Myoblast transplantation represents a promising therapeutic strategy in the treatment of several genetic muscular disorders including Duchenne muscular dystrophy. Nevertheless, such an approach is impaired by the rapid death, limited migration, and rejection of transplanted myoblasts by the host. Low molecular weight dextran sulfate (DXS), a sulfated polysaccharide, has been reported to act as a cytoprotectant for various cell types. Therefore, we investigated whether DXS could act as a “myoblast protectant” either in vitro or in vivo after transplantation in immunodeficient mice. In vitro, DXS bound human myoblasts in a dose dependent manner and significantly inhibited staurosporine-mediated apoptosis and necrosis. DXS pretreatment also protected human myoblasts from natural killer cell-mediated cytotoxicity. When human myoblasts engineered to express the renilla luciferase transgene were transplanted in immunodeficient mice, bioluminescence imaging analysis revealed that the proportion of surviving myoblasts 1 and 3 days after transplantation was two times higher when cells were preincubated with DXS compared to control (77.9 ± 10.1% vs. 39.4 ± 4.9%, p = 0.0009 and 38.1 ± 8.5% vs. 15.1 ± 3.4%, p = 0.01, respectively). Taken together, we provide evidence that DXS acts as a myoblast protectant in vitro and is able in vivo to prevent the early death of transplanted myoblasts.

Key words: Myoblast transplantation; Cell survival; Dextran sulfate (DXS); Bioluminescence; Mice

Received August 12, 2011; final acceptance May 26, 2012. Online prepub date: October 1, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Thomas Laumonier, Ph.D., Orthopaedic Surgery Service Geneva University Hospitals and Faculty of Medicine, 4, Rue Gabrielle Perret-Gentil, 1211 Geneva 14, Switzerland. Tel: +41 22 379 5393; Fax: +41 22 379 5402; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1227-1235, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657297
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Radio Electric Conveyed Fields Directly Reprogram Human Dermal Skin Fibroblasts Toward Cardiac, Neuronal, and Skeletal Muscle-Like Lineages

Margherita Maioli,*† Salvatore Rinaldi,‡ Sara Santaniello,*† Alessandro Castagna,‡ Gianfranco Pigliaru,*† Sara Gualini,*† Claudia Cavallini,†§ Vania Fontani,‡ and Carlo Ventura†§

*Department of Biomedical Sciences, University of Sassari, Sassari, Italy
†Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Bologna, Italy
‡Rinaldi Fontani Institute, Florence, Italy
§Cardiovascular Department, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy

Somatic cells can be directly reprogrammed to alternative differentiated fates without first becoming stem/progenitor cells. Nevertheless, the initial need for viral-mediated gene delivery renders this strategy unsafe in humans. Here, we provide evidence that exposure of human skin fibroblasts to a Radio Electric Asymmetric Conveyer (REAC), an innovative device delivering radio electric conveyed fields at a radiofrequency of 2.4 GHz, afforded remarkable commitment toward cardiac, neuronal, and skeletal muscle lineages. REAC induced the transcription of tissue-restricted genes, including Mef2c, Tbx5, GATA4, Nkx2.5, and prodynorphin for cardiac reprogramming, as well as myoD, and neurogenin 1 for skeletal myogenesis and neurogenesis, respectively. Conversely, REAC treatment elicited a biphasic effect on a number of stemness-related genes, leading to early transcriptional increase of Oct4, Sox2, cMyc, Nanog, and Klf4 within 6–20 h, followed by a downregulation at later times. The REAC action bypassed a persistent reprogramming toward an induced pluripotent stem cell-like state and involved the transcriptional induction of the NADPH oxidase subunit Nox4. Our results show for the first time the feasibility of using a physical stimulus to afford the expression of pluripotentiality in human adult somatic cells up to the attainment of three major target lineages for regenerative medicine.

Key words: Direct reprogramming; Human fibroblasts; Radio electric conveyed fields (RECF)

Received November 24, 2011; final acceptance May 31, 2012. Online prepub date: October 2, 2012.

Address correspondence to Prof. Carlo Ventura, Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures

and Biosystems, Strada Maggiore 42, 40125 Bologna, Italy. Tel/fax: +39-051-340339; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1237-1247, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657422
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Postnatal Transplantation of Interneuronal Precursor Cells Decreases Anxiety-Like Behavior in Adult Mice

M. F. Valente,* S. Romariz,* M. E. Calcagnotto,† L. Ruiz,* L. E. Mello,* R. Frussa-Filho,‡ and B. M. Longo*

*Department of Physiology, Universidade Federal de São Paulo, São Paulo, Brazil
†Department of Biochemistry, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil
‡Department of Pharmacology, Universidade Federal de São Paulo, São Paulo, Brazil

The GABAergic system is critically involved in the modulation of anxiety levels, and dysfunction of GABAergic neurotransmission appears to be involved in the development of generalized anxiety disorder. Precursor cells from the medial ganglionic eminence (MGE) have the ability to migrate and differentiate into inhibitory GABAergic interneurons after being transplanted into the mouse brain. Thus, transplantation of interneuronal precursor cells derived from the MGE into a postnatal brain could modify the neuronal circuitry, increasing GABAergic tone and decreasing anxiety-like behavior in animals. Our aim was to verify the in vivo effects of transplanted MGE cells by evaluating anxiety-like behavior in mice. MGE cells from 14-day green fluorescent protein (GFP) embryos were transplanted into newborn mice. At 15, 30, and 60 days posttransplant, the animals were tested for anxiety behavior with the elevated plus maze (EPM) test. Our results show that transplanted cells from MGE were able to migrate to different regions of the brain parenchyma and to differentiate into inhibitory interneurons. The neuronal precursor cell transplanted animals had decreased levels of anxiety, indicating a specific function of these cells in vivo. We suggested that transplantation of MGE-derived neuronal precursors into neonate brain could strengthen the inhibitory function of the GABAergic neuronal circuitry related to anxiety-like behavior in mice.

Key words: GABAergic neuron (GABA); Medial ganglionic eminence (MGE); Green fluorescent protein (GFP); Cell differentiation; Anxiolysis

Received August 11, 2011; final acceptance June 22, 2012. Online prepub date: October 1, 2012.
Address correspondence to Beatriz Monteiro Longo, Neurofisiologia, Depto. Fisiologia - UNIFESP - SP, R. Botucatu, 862 5 andar, V. Clementino - São Paulo - SP, Brazil. Tel: (5511) 5579-2033 or 5576-4513; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1249-1261, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X656153
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Embryonic-Derived Olfactory Ensheathing Cells Remyelinate Focal Areas of Spinal Cord Demyelination More Efficiently Than Neonatal or Adult-Derived Cells

David J. C. Coutts,*1 Charlotte E. Humphries,†1 Chao Zhao,* Giles W. Plant,† and Robin J. M. Franklin*

*Wellcome Trust-MRC Cambridge Stem Cell Institute and Department of Veterinary Medicine, University of Cambridge, Cambridge, UK
†Eileen Bond Spinal Cord Research Center, School of Anatomy and Human Biology, The University of Western Australia, Perth, Western Australia, Australia

Transplanted olfactory ensheathing cells (OECs) contribute to functional recovery in a range of CNS injuries by several mechanisms, one of which is potentially their ability to form myelin sheaths. OECs sourced from donors of different ages have been shown to remyelinate in several in vitro and in vivo models. However, the optimal donor age for OEC associated remyelination is unclear. This project directly compared the remyelinating potential of p75 purified OEC transplants from three donor ages. OECs were sourced from the olfactory bulbs of embryonic, neonatal, and adult rats and purified by immunopanning, and their remyelinating potential was directly compared by transplantation into the same adult rat toxin-induced model of spinal cord demyelination. Remyelination efficiency 3 weeks after transplantation was assessed morphologically and by immunostaining. Our results indicate that all donor ages remyelinate; however, this process is most efficiently achieved by embryonic-derived OECs.

Key words: Remyelination; Olfactory ensheathing cells (OECs); Transplantation; Age

Received November 29, 2011; final acceptance June 5, 2012. Online prepub date: October 2, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Robin J. M. Franklin, Wellcome Trust-MRC Cambridge Stem Cell Institute and Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Giles W. Plant, Stanford Partnership for Spinal Cord Injury and Repair, Stanford Institute for Neuroinnovation and Translational Neurosciences, Department of Neurosurgery, Lorry I Lokey Stem Cell Building, 265 Campus Drive, Stanford, CA 94305-5454, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1263-1279, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657242
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Stem Cell Grafting Improves Both Motor and Cognitive Impairments in a Genetic Model of Parkinson’s Disease, the Aphakia (ak) Mouse

Jisook Moon,*† Hyun-Seob Lee,† Jun Mo Kang,† Junpil Park,* Amanda Leung,* Sunghoi Hong,*1 Sangmi Chung,* and Kwang‑Soo Kim*

*Molecular Neurobiology Laboratory, McLean Hospital/Harvard Medical School, Belmont, MA, USA
†Department of Bioengineering, College of Life Science, CHA University, Seoul, Korea

Stem cell-based cell replacement of lost midbrain dopamine (mDA) neurons is a potential therapy for Parkinson’s disease (PD). Toward this goal, it is critical to optimize various aspects of cell transplantation and to assess functional recovery through behavioral tests in validated animal model(s) of PD. At present, cell transplantation studies are being done almost exclusively in neurotoxin-based animal models, because few genetic models of PD exhibit robust mDA neuronal loss. Here we used a genetic model of PD, the aphakia mouse, which demonstrates selective degeneration of mDA neurons in the substantia nigra. We systematically investigated the functional effects of transplanting embryonic stem cell-derived cells at different stages of in vitro differentiation: embryoid body (EB), neural progenitor (NP), and neuronal differentiated (ND) stages. We found that transplantation of NP cells yielded the best outcomes for both survival and behavioral improvement, while transplantation of EB and ND cells resulted in high teratoma-like tumor formation and poor survival, respectively. In behavioral paradigms specific to basal ganglia, the NP cells group prominently improved motor behavioral defects 1 and 2 months posttransplantation. Furthermore, we found that NP cell transplantation also improved cognitive impairments of aphakia mice, as examined by the passive avoidance task. Importantly, these graft-induced functional improvements well correlated with survival of tyrosine hydroxylase-positive DA neurons. Taken together, we propose that the aphakia mouse can serve as a novel and useful platform for cell transplantation studies to assess both neurological and cognitive improvements and that NP stage cells represent an optimal stage for transplantation.

Key words: Parkinson’s disease (PD); Dopaminergic neurons; Embryonic stem cells (ESCs); 6-Hydroxydopamine (6-OHDA); Aphakia (ak) mouse; Tyrosine hydroxylase (TH); l-3,4-Dihydroxyphenylalanine (l-DOPA)

Received July 9, 2011; final acceptance May 18, 2012. Online prepub date: October 2, 2012.
1Current address: Department of Biomedical Science, Korea University, Seoul, Korea.
Address correspondence to Kwang-Soo Kim, Ph.D., McLean Hospital/Harvard Medical School, 115 Mill Street, Belmont, MA 02178, USA. Tel: +1-617-855-2024; Fax: +1-617-855-3479; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Jisook Moon, Ph.D., Department of Bioengineering, CHA University, 606-16 Yeoksam-1 dong, Gangnam-gu, Seoul, South Korea. Tel: +82-2-3468-3196; Fax: +82-2-538-4102; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1281-1293, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X654984
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Survival and Functional Restoration of Human Fetal Ventral Mesencephalon Following Transplantation in a Rat Model of Parkinson’s Disease

Anika Rath,*1 Alexander Klein,†1 Anna Papazoglou,* Jan Pruszak,‡ Joanna Garcia,*§ Martin Krause,* Jaroslaw Maciaczyk,* Stephen B. Dunnett,† and Guido Nikkhah*

*Department of Stereotactic and Functional Neurosurgery, Neurocentre, University of Freiburg, Freiburg, Germany
†Brain Repair Group, School of Biosciences, Cardiff University, Cardiff, Wales, UK
‡Institute for Anatomy and Cell Biology, University of Freiburg, Freiburg, Germany
§Department of Neurology, Neurocentre, University of Freiburg, Freiburg, Germany

Cell replacement therapy by intracerebral transplantation of fetal dopaminergic neurons has become a promising therapeutic option for patients suffering from Parkinson’s disease during the last decades. However, limited availability of human fetal tissue as well as ethical issues, lack of alternative nonfetal donor cells, and the absence of standardized transplantation protocols have prevented neurorestorative therapies from becoming a routine procedure in patients suffering from neurodegenerative diseases. Improvement of graft survival, surgery techniques, and identification of the optimal target area are imperative for further optimization of this novel treatment. In the present study, human primary fetal ventral mesencephalon-derived tissue from 7- to 9-week-old human fetuses was transplanted into 6-hydroxydopamine-lesioned adult Sprague–Dawley rats. Graft survival, fiber outgrowth, and drug-induced rotational behavior up to 14 weeks posttransplantation were compared between different intrastriatal transplantation techniques (full single cell suspension vs. partial tissue pieces suspension injected by glass capillary or metal cannula) and the intranigral glass capillary injection of a full (single cell) suspension. The results demonstrate a higher survival rate of dopamine neurons, a greater reduction in amphetamine-induced rotations (overcompensation), and more extensive fiber outgrowth for the intrastriatally transplanted partial (tissue pieces) suspension compared to all other groups. Apomorphine-induced rotational bias was significantly reduced in all groups including the intranigral group. The data confirm that human ventral mesencephalon-derived cells serve as a viable cell source, survive in a xenografting paradigm, and functionally integrate into the host tissue. In contrast to rat donor cells, keeping the original (fetal) neuronal network by preparing only a partial suspension containing tissue pieces seems to be beneficial for human cells, although a metal cannula that causes greater tissue trauma to the host is required for injection. In addition, homotopic intranigral grafts may represent a complimentary grafting approach to the “classical” ectopic intrastriatal target site in PD.

Key words: Neurorestoration; Single cell suspension; Tissue pieces; Metal cannula; Glass capillary

Received July 8, 2011; final acceptance February 20, 2012. Online prepub date: September 7, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Guido Nikkhah, Department of Stereotactic and Functional Neurosurgery, Neurocentre, University of Freiburg, Breisacher Str. 64, 79106 Freiburg, Germany. Tel: +49 761 27050630; Fax: +49 761 27050100; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1295-1305, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658683
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

PET Imaging of Serotonin Transporters With 4-[18F]-ADAM in a Parkinsonian Rat Model

Shao-Ju Weng,*† Chyng-Yann Shiue,‡ Wen-Sheng Huang,§ Cheng-Yi Cheng,‡ San-Yuan Huang,¶ I-Hsun Li,# Chih-Chieh Tao,† Ta-Kai Chou,‡ Mei-Hsiu Liao,** Yung-Ping Chang,†† and Kuo-Hsing Ma†

*Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan
†Department of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan
‡Department of Nuclear Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
§Departments of Nuclear Medicine and Medical Research, Changhua Christian Hospital, Changhua, Taiwan
¶Department of Psychiatry, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan
#School of Pharmacy, National Defense Medical Center, Taipei, Taiwan
**Institute of Nuclear Energy Research, Taoyaun, Taiwan
††Department of Pharmacy, Songshan Armed Forces General Hospital, Taipei, Taiwan

This study was undertaken to address the effects of fetal mesencephalic tissue transplantation on the serotonin system in a rat model of Parkinson’s disease (PD) while also investigating the usefulness of 4-[18F]-ADAM (a serotonin transporter imaging agent) coupled with micro-PET for imaging serotonin transporters (SERTs). A PD model was induced by unilateral injection of 6-hydroxydopamine (6-OHDA) into the right medial forebrain bundle of the nigrostriatal pathway, while cell transplantation was performed via intrastriatal injection of mesencephalic brain tissue dissected from embryonic (E14) rats. The 4-[18F]-ADAM/micro-PET scanning was performed following both 6-OHDA lesioning and transplantation. Immunohistochemistry (IHC) studies were also performed following the final PET scan, and the results were compared to show a 17–43% decrease in the specific uptake ratio (SUR) and a 23–52% decrease in serotonin transporter immunoreactivity (SERT-ir) within various brain regions on the lesioned side. The number of methamphetamine-induced rotations also decreased significantly at the 4th week postgraft. In addition, striatal SUR and the SERT-ir levels were restored to 77% and 83% 5 weeks postgraft. These results suggest that Parkinson’s disease also affects the serotonergic system, while both the dopaminergic and serotonergic systems can be partially restored in a rat model of PD after E14 mesencephalic tissue transplantation. In addition, we have also determined that 4-[18F]-ADAM/micro-PET can be used to detect serotonergic neuron loss, monitor the progress of Parkinson’s disease, and oversee the effectiveness of therapy.

Key words: Parkinson’s disease (PD); Rat model; Mesencephalic tissue; Serotonin transporter (SERT); 4-[18F]-ADAM

Received February 24, 2011; final acceptance May 6, 2012. Online prepub date: October 31, 2012.
Address correspondence to Kuo-Hsing Ma, Ph.D., Department of Biology and Anatomy, National Defense Medical Center, Taipei, No. 161, Sec. 6, Min-Chuan E. Road, Neihu 114, Taipei, Taiwan. Tel: +886-2-87923100, ext. 18160; Fax: +886-2-87923159; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it