Cell Transplantation 21 (6) Abstracts

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Cell Transplantation, Vol. 21, pp. 1065–1074, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X612477
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review
Lessons From Genetically Altered Mesenchymal Stem Cells (MSCs): Candidates for Improved MSC-Directed Myocardial Repair

Maria P. Alfaro* and Pampee P. Young*†

*Department of Pathology, Vanderbilt University Medical Center, Nashville, TN, USA
†Department of Veterans Affairs Medical Center, Nashville, TN, USA

The regenerative and reparative potential of mesenchymal stem cells (MSCs) make them attractive candidates for numerous cell-directed therapies. The variant degree of tissue repair by transplanted MSCs has been assessed in several published reports. There are many gaps in the knowledge of MSC biology and the underlying reasons for their disparate effectiveness in tissue repair. This review examines successful preclinical models of MSC-directed repair, particularly of myocardial repair, in an attempt to shed light into the events dictating MSC therapeutic efficacy. The reparative advantage of genetically altered MSCs will be described. This overview will elucidate possible molecular mechanisms that can influence MSC engraftment, differentiation, self-renewal, and ultimately increase wound repair.

Key words: Mesenchymal stem cells (MSCs); Self-renewal; Transplantation; Cell biology

Received March 14, 2011; final acceptance June 13, 2011. Online prepub date: November 11, 2011.
Address correspondence to Pampee P. Young, Department of Pathology, Vanderbilt University School of Medicine, 1161 21st Ave. South, C2217A MCN, Nashville, TN 37232, USA. Fax: 615-343-7023; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1075–1085, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640538
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Hair Follicle Stem Cells Derived From Single Rat Vibrissa Via Organ Culture Reconstitute Hair Follicles In Vivo

Shoubing Zhang,*†1 Huimin Hu,*†1 Huishan Zhang,*†1 Shuang Liu,*† Shu Liu,*† Ying Zhang,*† Xiaohua Lei,* Lina Ning,* Yujing Cao,* and Enkui Duan*

*State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China
†Graduate University of the Chinese Academy of Sciences, Beijing, China

Hair follicle stem cells (HFSCs) are potentially useful for the treatment of skin injuries and diseases. To achieve clinical application, a prerequisite must be accomplished: harvesting enough HFSCs from limited skin biopsy. The commonly used sorting approach for isolating HFSCs, however, suffers from its intrinsic disadvantages, such as requirement of large-scale skin biopsy. Here, we report an efficient organ culture method to isolate and expand rat HFSCs from limited skin biopsy and these HFSCs could reconstitute the epidermis and the hair follicles (HFs). Seventy-three percent of cultured HFs formed hair follicle stem cell colonies from the bulge, and a single hair follicle provided all the HFSCs used in this research, demonstrating the high efficiency of this method. Quantitative RT-PCR and immunofluorescent staining results revealed that these stem cells obtained from the bulge highly expressed basal layer markers K14 and alpha-6 integrin, epithelial stem cell marker P63, and bulge stem cell marker K15. After long-term culture in vitro, GFP-labeled hair follicle stem cells formed new hair follicles, epidermis, and sebaceous glands following xenotransplantation into the back of nude mice. This study indicated that multipotent hair follicle stem cells could be efficiently harvested through organ culture from limited skin material—even a single hair follicle—and reconstitute hair follicles in vivo after long-term expansion culture, providing the basis for future clinical applications.

Key words: Hair follicle stem cell (HFSC); Organ culture; Bulge; Hair follicle reconstitution

Received January 6, 2011; final acceptance June 30, 2011. Online prepub date: April 20, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Enkui Duan, State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1087–1093, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636876
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Isolation of Myogenic Stem Cells From Cultures of Cryopreserved Human Skeletal Muscle

Bo Zheng,*1 Chien-Wen Chen,*†1 Guangheng Li,* Seth D. Thompson,* Minakshi Poddar,* Bruno Pe´ault,*‡§¶ and Johnny Huard*†

*Stem Cell Research Center, Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine and Children’s Hospital of Pittsburgh of UPMC, Pittsburgh, PA, USA
†Department of Bioengineering, University of Pittsburgh Swanson School of Engineering, Pittsburgh, PA, USA
‡Department of Pediatrics, Children’s Hospital of Pittsburgh of UPMC, Pittsburgh, PA, USA
§David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA, USA
¶Center for Vascular Science and Center for Regenerative Medicine, University of Edinburgh, Edinburgh, UK

We demonstrate that subpopulations of adult human skeletal muscle-derived stem cells, myogenic endothelial cells (MECs), and perivascular stem cells (PSCs) can be simultaneously purified by fluorescence-activated cell sorting (FACS) from cryopreserved human primary skeletal muscle cell cultures (cryo-hPSMCs). For FACS isolation, we utilized a combination of cell lineage markers: the myogenic cell marker CD56, the endothelial cell marker UEA-1 receptor (UEA-1R), and the perivascular cell marker CD146. MECs expressing all three cell lineage markers (CD56+UEA-1R+CD146+/CD45) and PSCs expressing only CD146 (CD146+/CD45CD56UEA-1R) were isolated by FACS. To evaluate their myogenic capacities, the sorted cells, with and without expansion in culture, were transplanted into the cardiotoxin-injured skeletal muscles of immunodeficient mice. The purified MECs exhibited the highest regenerative capacity in the injured mouse muscles among all cell fractions tested, while PSCs remained superior to myoblasts and the unpurified primary skeletal muscle cells. Our findings show that both MECs and PSCs retain their high myogenic potentials after in vitro expansion, cryopreservation, and FACS sorting. The current study demonstrates that myogenic stem cells are prospectively isolatable from long-term cryopreserved primary skeletal muscle cell cultures. We emphasize the potential application of this new approach to extract therapeutic stem cells from human muscle cells cryogenically banked for clinical purposes.

Key words: Myogenesis; Human skeletal muscle; Myogenic endothelial cells (MECs); Perivascular stem cells (PSCs); Cell therapy

Received July 12, 2010; final acceptance July 10, 2011. Online prepub date: March 21, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Johnny Huard, Ph.D., Stem Cell Research Center, Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, 206 Bridgeside Point II, 450 Technology Drive, Pittsburgh, PA, USA 15219, USA. Tel: (412) 648-2798; Fax: (412) 648-4066; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1095–1107, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X623880
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Pretreatment of Endothelial Progenitor Cells With Osteopontin Enhances Cell Therapy for Peripheral Vascular Disease

E. E. Vaughan,*1 A. Liew,*1 K. Mashayekhi,* P. Dockery,† J. McDermott,* B. Kealy,* A. Flynn,* A. Duffy,‡ C. Coleman,* A. O’Regan,§ F. P. Barry,* and T. O’Brien*

*Regenerative Medicine Institute (REMEDI), National Centre for Biomedical Engineering Science (NCBES), National University Ireland Galway (NUIG), Galway, Ireland
†Department of Anatomy, National University Ireland Galway (NUIG), Galway, Ireland
‡Medtronic, Galway, Ireland
§Department of Respiratory Medicine, University College Hospital Galway, Galway, Ireland

Tissue necrosis resulting from critical limb ischemia (CLI) leads to amputation in a significant number of patients. Autologous cell therapy using angiogenic cells such as endothelial progenitor cells (EPCs) holds promise as a treatment for CLI but a limitation of this treatment is that the underlying disease etiology that resulted in CLI may also contribute to dysfunction of the therapeutic EPCs. This study aimed to elucidate the mechanism of EPC dysfunction using diabetes mellitus as a model and to determine whether correction of this defect in dysfunctional EPCs ex vivo would improve the outcome after cell transplantation in the murine hind limb ischemia model. EPC dysfunction was confirmed in a homogenous population of patients ith type 1 diabetes mellitus and a microarray study was preformed to identify dysregulated genes. Notably, the secreted proangiogenic protein osteopontin (OPN) was significantly downregulated in diabetic EPCs. Furthermore, OPN-deficient mice showed impaired recovery following hind limb ischemia, suggesting a critical role for OPN in postnatal neovascularization. EPCs isolated from OPN KO mice showed decreased ability to adhere to endothelial cells as well as impaired angiogenic potential. However, this dysfunction was reversed upon exposure to recombinant OPN, suggesting that OPN may act in an autocrine manner on EPCs. Indeed, exposure of OPN knockout (KO) EPCs to OPN was sufficient to induce the secretion of angiogenic proteins (IL-6, TGF-α, and FGF-α). We also demonstrated that vascular regeneration following hind limb ischemia in OPN KO mice was significantly improved upon injection of EPCs preexposed to OPN. We concluded that OPN acts in an autocrine manner on EPCs to induce the secretion of angiogenic proteins, thereby playing a critical role in EPC-mediated neovascularization. Modification of cells by exposure to OPN may improve the efficacy of autologous EPC transplantation via the enhanced secretion of angiogenic proteins.

Key words: Endothelial progenitor cell (EPC); Stem cell; Diabetes mellitus; Osteopontin; Vascular disease; Ischemia

Received September 29, 2010; final acceptance June 10, 2011. Online prepub date: February 2, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Prof. Timothy O’Brien, Regenerative Medicine Institute (REMEDI), National Centre for Biomedical Engineering Science (NCBES), National University Ireland Galway (NUIG), University Road, Galway, Ireland. Tel: +353 91-495107; Fax: +353 91-495547; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1109–1120, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X603657
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Percutaneous Adventitial Delivery of Allogeneic Bone Marrow-Derived Stem Cells Via Infarct-Related Artery Improves Long-Term Ventricular Function in Acute Myocardial Infarction

Satish Medicetty,*1 Dominik Wiktor,†1 Nicholas Lehman,* Amy Raber,* Zoran B. Popovic,† Robert Deans,* Anthony E. Ting,*2 and Marc S. Penn†2

*Regenerative Medicine Department, Athersys, Inc., Cleveland, OH, USA

†Skirball Laboratory for Cardiovascular Cellular Therapeutics, Departments of Stem Cell Biology and Regenerative Medicine and Cardiovascular Medicine, Cleveland Clinic, Cleveland, OH, USA

Acute myocardial infarction (AMI) results in ischemic damage and death of cardiomyocytes and loss of vasculature. Stem cell therapy has emerged as a potentially promising strategy for maximizing cardiac function following ischemic injury. Issues of cell source, delivery, and quantification of response have challenged development of clinically viable strategies. In this study we investigate the effects of a well-defined bone marrow-derived allogeneic cell product delivered by catheter directly to the myocardium via the infarct-related vessel on global and regional measures of left ventricular (LV) function in a porcine model of anterior wall myocardial infarction. Multipotent adult progenitor cells (MAPCs) were derived and expanded from the bone marrow of a donor Yorkshire pig. Anterior wall myocardial infarction (AMI) was induced by 90 min of mid-LAD occlusion using a balloon catheter. Two days after AMI was induced, either vehicle (Plasma Lyte-A, n = 7), low-dose (20 million, n = 6), or high-dose (200 million, n = 6) MAPCs were delivered directly to the myocardium via the infarct-related vessel using a transarterial microsyringe catheter-based delivery system. Echocardiography was used to measure LV function as a function of time after AMI. Animals that received low-dose cell treatment showed significant improvement in regional and global LV function and remodeling compared to the high-dose or control animals. Direct myocardial delivery of allogeneic MAPCs 2 days following AMI through the vessel wall of the infarct-related vessel is safe and results in delivery of cells throughout the infarct zone and improved cardiac function despite lack of long-term cell survival. These data further support the hypothesis of cell-based myocardial tissue repair by a paracrine mechanism and suggest a clinically translatable strategy for delivering cells at any time after AMI to modulate cardiac remodeling and function.

Key words: Acute myocardial infarction (AMI); Multipotent adult progenitor cells (MAPCs); Adventitial delivery

Received June 29, 2010; final acceptance July 13, 2011. Online prepub date: October 14, 2011.
1These authors provided equal contribution to this work.
2These authors provided equal contribution to this work.
Address correspondence to Marc S. Penn, M.D., Ph.D., at his current address: Summa Cardiovascular Institute, Summa Health System, 525 E. Market St., Akron, OH 44309, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1121–1135, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627381
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Magnetic Enhancement of Cell Retention, Engraftment, and Functional Benefit After Intracoronary Delivery of Cardiac-Derived Stem Cells in a Rat Model of Ischemia/Reperfusion

Ke Cheng,* Konstantinos Malliaras,* Tao-Sheng Li,*† Baiming Sun,* Christiane Houde,* Giselle Galang,* Jeremy Smith,* Noriko Matsushita,* and Eduardo Marbán*

*Cedars-Sinai Heart Institute, Los Angeles, CA, USA
†Department of Stem Cell Biology, Nagasaki University Graduate School of Biomedical Science, Nagasaki, Japan

The efficiency of stem cell transplantation is limited by low cell retention. Intracoronary (IC) delivery is convenient and widely used but exhibits particularly low cell retention rates. We sought to improve IC cell retention by magnetic targeting. Rat cardiosphere-derived cells labeled with iron microspheres were injected into the left ventricular cavity of syngeneic rats during brief aortic clamping. Placement of a 1.3 Tesla magnet ~1 cm above the heart during and after cell injection enhanced cell retention at 24 h by 5.2–6.4-fold when 1, 3, or 5 × 105 cells were infused, without elevation of serum troponin I (sTnI) levels. Higher cell doses (1 or 2 × 106 cells) did raise sTnI levels, due to microvascular obstruction; in this range, magnetic enhancement did not improve cell retention. To assess efficacy, 5 × 105 iron-labeled, GFP-expressing cells were infused into rat hearts after 45 min ischemia/20 min reperfusion of the left anterior coronary artery, with and without a superimposed magnet. By quantitative PCR and optical imaging, magnetic targeting increased cardiac retention of transplanted cells at 24 h, and decreased migration into the lungs. The enhanced cell engraftment persisted for at least 3 weeks, at which time left ventricular remodeling was attenuated, and therapeutic benefit (ejection fraction) was higher, in the magnetic targeting group. Histology revealed more GFP+ cardiomyocytes, Ki67+ cardiomyocytes and GFP/ckit+ cells, and fewer TUNEL+ cells, in hearts from the magnetic targeting group. In a rat model of ischemia/reperfusion injury, magnetically enhanced intracoronary cell delivery is safe and improves cell therapy outcomes.

Key words: Cardiac stem cells; Targeted cell delivery; Myocardial infarction

Received October 15, 2010; final acceptance August 6, 2011. Online prepub date: March 8, 2012.
Address correspondence to Eduardo Marba´n, M.D., Ph.D., Cedars-Sinai Heart Institute, 8700 Beverly Blvd, 1090 Davis Research Building, Los Angeles, CA 90048, USA. Tel: 310 423 7557; Fax: 310 423 7637; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Ke Cheng, Ph.D., Cedars-Sinai Heart Institute, 8700 Beverly Blvd., 1090 Davis Research Building, Los Angeles, CA 90048, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1137–1148, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627435
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Focused Magnetic Stem Cell Targeting to the Retina Using Superparamagnetic Iron Oxide Nanoparticles

Anat Yanai,* Urs O. Häfeli,† Andrew L. Metcalfe,* Peter Soema,† Lois Addo,* Cheryl Y. Gregory-Evans,* Kelvin Po,* Xianghong Shan,* Orson L. Moritz,* and Kevin Gregory-Evans*

*Department of Ophthalmology & Visual Science, Faculty of Medicine, University of British Columbia, Vancouver, Canada
†Faculty of Pharmaceutical Sciences, University of British Columbia,Vancouver, Canada

Developing new ways of delivering cells to diseased tissue will be a key factor in translating cell therapeutics research into clinical use. Magnetically targeting cells enables delivery of significant numbers of cells to key areas of specific organs. To demonstrate feasibility in neurological tissue, we targeted cells magnetically to the upper hemisphere of the rodent retina. Rat mesenchymal stem cells (MSCs) were magnetized using superparamagnetic iron oxide nanoparticles (SPIONs). In vitro studies suggested that magnetization with fluidMAG-D was well tolerated, that cells remained viable, and they retained their differentiation capabilities. FluidMAG-D-labeled MSCs were injected intravitreally or via the tail vein of the S334ter-4 transgenic rat model of retinal degeneration with or without placing a gold-plated neodymium disc magnet within the orbit, but outside the eye. Retinal flatmount and cryosection imaging demonstrated that after intravitreal injection cells localized to the inner retina in a tightly confined area corresponding to the position of the orbital magnet. After intravenous injection, similar retinal localization was achieved and remarkably was associated with a tenfold increase in magnetic MSC delivery to the retina. Cryosections demonstrated that cells had migrated into both the inner and outer retina. Magnetic MSC treatment with orbital magnet also resulted in significantly higher retinal concentrations of anti-inflammatory molecules interleukin-10 and hepatocyte growth factor. This suggested that intravenous MSC therapy also resulted in significant therapeutic benefit in the dystrophic retina. With minimal risk of collateral damage, these results suggest that magnetic cell delivery is the best approach for controlled delivery of cells to the outer retina—the focus for disease in age-related macular degeneration and retinitis pigmentosa.

Key words: Magnetic targeting; Retina; Superparamagnetic iron oxide nanoparticles (SPIONs)

Received January 12, 2011; final acceptance July 14, 2011. Online prepub date: March 8, 2012.
Address correspondence to Kevin Gregory-Evans, M.D., Ph.D., FRCS, FRCOphth., Centre for Macular Research, Department Ophthalmology and Visual Sciences, University of British Columbia, 2550 Willow Street, Vancouver, BC, Canada V5Z 3N9. Tel: +1-604-671-0419; Fax: +1-604-875-4663; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1149–1159, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X593154
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Effects of Myeloablation, Peripheral Chimerism, and Whole-Body Irradiation on the Entry of Bone Marrow-Derived Cells Into the Brain

Antoine Lampron, Martine Lessard, and Serge Rivest

Laboratory of Endocrinology and Genomics, CHUL Research Center and Department of Molecular Medicine, Faculty of Medicine, Laval University, Québec, Canada

Understanding how bone marrow-derived cells (BMDCs) enter the central nervous system (CNS) is critical for the development of therapies for brain-related disorders using hematopoietic stem cells. We investigated the brain damages and blood–brain barrier (BBB) modification following either whole-body irradiation or a myeloablative chemotherapy regimen in mice, and the capacity for these treatments to induce the entry of BMDCs into the CNS. Neither treatment had a lasting effect on brain integrity and both were equally efficient at achieving myeloablation. Injection of bone marrow cells from green fluorescent protein (GFP) transgenic mice was able to completely repopulate the hematopoietic niche in the circulation and in hematopoietic organs (thymus and spleen). However, GFP+ cells only entered the brain following whole-body irradiation. We conclude that myeloablation, damages to the brain integrity, or the BBB and peripheral chimerism are not responsible for the entry of BMDCs into the CNS following irradiation.

Key words: Microglia; Central nervous sysem (CNS); Neuroimmunology; Hematopoietic stem cell; Irradiation; Chemotherapy; Bone marrow-derived cells; Innate immunity

Received February 22, 2011; final acceptance June 8, 2011. Online prepub date: September 22, 2011.
Address correspondence to Dr. Serge Rivest, Laboratory of Endocrinology and Genomics, CHUL Research Center and Department of Molecular Medicine, Faculty of Medicine, Laval University, Québec, Canada. Tel: (418) 654-2296; Fax: (418) 654-2761; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1161–1175, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627444
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Highly Malignant Behavior of a Murine Oligodendrocyte Precursor Cell Line Following Transplantation Into the Demyelinated and Nondemyelinated Central Nervous System

Florian Hansmann,*†1 Kidsadagon Pringproa,*†1 Reiner Ulrich,* Yanyong Sun,*† Vanessa Herder,*† Mihaela Kreutzer,*† Wolfgang Baumgärtner,*† and Konstantin Wewetzer*†‡

*Department of Pathology, University of Veterinary Medicine Hannover, Hannover, Germany
†Centre for Systems Neuroscience Hannover, Hannover, Germany
‡Centre of Anatomy, Department of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany

Understanding the basic mechanisms that control CNS remyelination is of direct clinical relevance. Suitable model systems include the analysis of naturally occurring and genetically generated mouse mutants and the transplantation of oligodendrocyte precursor cells (OPCs) following experimental demyelination. However, aforementioned studies were exclusively carried out in rats and little is known about the in vivo behavior of transplanted murine OPCs. Therefore in the present study, we (i) established a model of ethidium bromide-induced demyelination of the caudal cerebellar peduncle (CCP) in the adult mouse and (ii) studied the distribution and marker expression of the murine OPC line BO-1 expressing the enhanced green fluorescent protein (eGFP) 10 and 17 days after stereotaxic implantation. Injection of ethidium bromide (0.025%) in the CCP resulted in a severe loss of myelin, marked astrogliosis, and mild to moderate axonal alterations. Transplanted cells formed an invasive and liquorogenic metastasizing tumor, classified as murine giant cell glioblastoma. Transplanted BO-1 cells displayed substantially reduced CNPase expression as compared to their in vitro phenotype, low levels of MBP and GFAP, prominent upregulation of NG2, PDGFRα, nuclear p53, and an unaltered expression of signal transducer and activator of transcription (STAT)-3. Summarized environmental signaling in the brain stem was not sufficient to trigger oligodendrocytic differentiation of BO-1 cells and seemed to block CNPase expression. Moreover, the lack of the remyelinating capacity was associated with tumor formation indicating that BO-1 cells may serve as a versatile experimental model to study tumorigenesis of glial tumors.

Key words: Caudal cerebellar peduncle; Murine giant cell glioblastoma; Oligodendrocyte precursor cell; Signal transducer and activator of transcription-3; Transplantation

Received October 28, 2010; final acceptance June 8, 2011. Online prepub date: March 8, 2012.
1These authors provided equal contribution and are considered as first authors.
Address correspondence to Prof. Dr. Wolfgang Baumgärtner, Ph.D., Dipl. ECVP, Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany. Tel: +49-511-953-8620; Fax: +49-511-953-8675; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1177–1197, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X612503
E-ISSN 1555-3892
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The Effect of Growth Factors and Soluble Nogo-66 Receptor Protein on Transplanted Neural Stem/Progenitor Survival and Axonal Regeneration After Complete Transection of Rat Spinal Cord

Xiaodong Guo,*† Tasneem Zahir,‡ Andrea Mothe,† Molly S. Shoichet,‡ Cindi M. Morshead,§ Yusuke Katayama,‡ and Charles H. Tator†

*Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China
†Toronto Western Research Institute, Toronto Western Hospital and University of Toronto, Toronto, Ontario, Canada
‡Department of Chemical Engineering & Applied Chemistry, Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada
§Department of Surgery and Institute of Medical Sciences, Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada

Adult central mammalian axons show minimal regeneration after spinal cord injury due to loss of oligodendrocytes, demyelination of surviving axons, absence of growth-promoting molecules, and inhibitors of axonal outgrowth. In the present study, we attempted to address these impediments to regeneration by using a combinatory strategy to enhance cell survival and regeneration after complete spinal cord transection (SCT) in adult rats. The strategy comprised: 1) adult rat brain-derived neural stem/progenitor cells (NSPCs) preseeded on laminin-coated chitosan channels; 2) extramedullary chitosan channels to promote axonal regrowth and reduce the barrier caused by scarring; 3) local delivery of a novel rat soluble Nogo-66 receptor protein [NgR(310)ecto-Fc, referred to as NgR] to block the inhibitory effect of myelin-based inhibitors; and 4) local delivery of basic fibroblast growth factor, epidermal growth factor, and platelet-derived growth factor to enhance survival and promote differentiation of transplanted cells. Compared with our previous studies where brain-derived NSPCs preseeded in extramedullary chitosan channels were implanted in the same SCT model but without growth factors and NgR, the present channel–growth factor combination produced greater numbers of surviving NSPCs after SCT. Also, the growth factors promoted preferential differentiation of NSPCs toward oligodendrocytes, while NgR significantly decreased astrocytic differentiation of NSPCs. NgR alone or in combination with NSPCs significantly enhanced the total number of myelinated fibers in the bridge and increased the area of the bridging tissue between the cord stumps. The combination of NgR, growth factors, and NSPCs had synergistic effect on bridge formation. However, only a small number of descending corticospinal tract axons grew into the central portions of the bridges as shown by anterograde tracing of the corticospinal tract with BDA. The majority of the regenerated axons in the channels originated from local host neurons adjacent to the tissue bridges. In conclusion, we showed that growth factors increased survival of transplanted NSPCs whereas NgR enhanced axonal regeneration, but the combination did not have additive effects on functional recovery or regeneration.

Key words: Spinal cord injury (SCI); Neural stem/progenitor cells (NSPCs); Nogo; Nogo-66 receptor; Chitosan channel; Axonal regeneration

Received October 7, 2009; final acceptance July 19, 2011. Online prepub date: January 10, 2012.
Address correspondence to Charles H. Tator, Ph.D., M.D., Toronto Western Research Institute, Toronto Western Hospital and University of Toronto, Room 12-435, McLaughlin Wing, 399 Bathurst St., Toronto, Ontario, Canada M5T 2S8. Tel: (416) 603-5889; Fax: (416) 603-5298; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1199–1211, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X589609
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Determination of the Therapeutic Time Window for Human Umbilical Cord Blood Mononuclear Cell Transplantation Following Experimental Stroke in Rats

Johannes Boltze,*†1 Uwe R. Schmidt,*1 Doreen M. Reich,* Alexander Kranz,*† Klaus G. Reymann,‡§ Maria Strassburger,‡ Donald Lobsien,¶ Daniel-Christoph Wagner,*† Annette Förschler,¶# and Wolf-Rüdiger Schäbitz**

*Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany
†Translational Centre for Regenerative Medicine, University of Leipzig, Leipzig, Germany
‡Leibniz Institute for Neurobiology, Project Group Neuropharmacology, Magdeburg, Germany
§German Centre for Neurodegenerative Diseases (DZNE), Magdeburg, Germany
¶Department for Neuroradiology, University of Leipzig, Leipzig, Germany
#Department of Neuroradiology, University Hospital rechts der Isar, Technical University of Munich, Munich, Germany
**Department of Neurology, Evangelisches Krankenhaus Bielefeld, Bielefeld, Germany

Experimental treatment strategies using human umbilical cord blood mononuclear cells (hUCB MNCs) represent a promising option for alternative stroke therapies. An important point for clinical translation of such treatment approaches is knowledge on the therapeutic time window. Although expected to be wider than for thrombolysis, the exact time window for hUCB MNC therapy is not known. Our study aimed to determine the time window of intravenous hUCB MNC administration after middle cerebral artery occlusion (MCAO). Male spontaneously hypertensive rats underwent MCAO and were randomly assigned to hUCB MNC administration at 4, 24, 72, and 120 or 14 days. Influence of cell treatment was observed by magnetic resonance imaging on days 1, 8, and 29 following MCAO and by assessment of functional neurological recovery. On day 30, brains were screened for glial scar development and presence of hUCB MNCs. Further, influence of hUCB MNCs on necrosis and apoptosis in postischemic neural tissue was investigated in hippocampal slices cultures. Transplantation within a 72-h time window resulted in an early improvement of functional recovery, paralleled by a reduction of brain atrophy and diminished glial scarring. Cell transplantation 120 h post-MCAO only induced minor functional recovery without changes in the brain atrophy rate and glial reactivity. Later transplantation (14 days) did not show any benefit. No evidence for intracerebrally localized hUCB MNCs was found in any treatment group. In vitro hUCB MNCs were able to significantly reduce postischemic neural necrosis and apoptosis. Our results for the first time indicate a time window of therapeutic hUCB MNC application of at least 72 h. The time window is limited, but wider than compared to conventional pharmacological approaches. The data furthermore confirms that differentiation and integration of administered cells is not a prerequisite for poststroke functional improvement and lesion size reduction.

Key words: Ischemic stroke; Human umbilical cord blood (hUCB); Time window; Behavioral phenotyping; Magnetic resonance imaging

Received November 26, 2010; final acceptance June 12, 2011. Online prepub date: December 13, 2011.
1These authors provided equal contribution to this work.
Address correspondence to Johannes Boltze, M.D., Ph.D., Fraunhofer-Institute for Cell Therapy and Immunology, Perlickstr.1, D-04103 Leipzig, Germany. Tel: +49-341-97-25814; Fax: +49-341-97-25829; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1213–1223, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X600948
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

CDNF Protects the Nigrostriatal Dopamine System and Promotes Recovery After MPTP Treatment in Mice

Mikko Airavaara,* Brandon K. Harvey,* Merja H. Voutilainen,† Hui Shen,* Jenny Chou,* Päivi Lindholm,‡ Maria Lindahl,‡ Raimo K. Tuominen,† Mart Saarma,‡ Barry Hoffer,* and Yun Wang*

*Intramural Research Program, National Institute on Drug Abuse, NIH, Baltimore, MD, USA
†Faculty of Pharmacy, Department of Pharmacology and Toxicology, University of Helsinki, Helsinki, Finland
‡Institute of Biotechnology, University of Helsinki, Helsinki, Finland

Cerebral dopamine neurotrophic factor (CDNF) is a recently discovered protein, which belongs to the evolutionarily conserved CDNF/MANF family of neurotrophic factors. The degeneration of dopamine neurons following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment is well characterized, and efficacy in this model is considered a standard criterion for development of parkinsonian therapies. MPTP is a neurotoxin, which produces parkinsonian symptoms in humans and in C57/Bl6 mice. To date, there are no reports about the effects of CDNF on dopamine neuron survival or function in the MPTP rodent model, a critical gap. Therefore, we studied whether CDNF has neuroprotective and neurorestorative properties for the nigrostriatal dopamine system after MPTP injections in C57/Bl6 mice. We found that bilateral striatal CDNF injections, given 20 h before MPTP, improved horizontal and vertical motor behavior. CDNF pretreatment increased tyrosine hydroxylase (TH) immunoreactivity in the striatum and in the substantia nigra pars reticulata (SNpr), as well as the number of TH-positive cells in substantia nigra pars compacta (SNpc). Posttreatment with CDNF, given 1 week after MPTP injections, increased horizontal and vertical motor behavior of mice, as well as dopamine fiber densities in the striatum and the number of TH-positive cells in SNpc. CDNF did not alter any of the analyzed dopaminergic biomarkers or locomotor behavior in MPTP-untreated animals. We conclude that striatal CDNF administration is both neuroprotective and neurorestorative for the TH-positive cells in the nigrostriatal dopamine system in the MPTP model, which supports the development of CDNF-based treatment for Parkinson’s disease.

Key words: Cerebral dopamine neurotrophic factor (CDNF); Mesencephalic astrocyte-derived neurotrophic factor (MANF); 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP); Parkinson’s disease

Received June 1, 2011; final acceptance June 30, 2011. Online prepub date: September 22, 2011.
Address correspondence to Yun Wang, Intramural Research Program, National Institute on Drug Abuse, 251 Bayview Blvd., Baltimore, MD 21224, USA. Tel: 443 812-6825; Fax: 443 740-2840; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1125–1133, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X600966
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Radiofrequency Energy Loop Primes Cardiac, Neuronal, and Skeletal Muscle Differentiation in Mouse Embryonic Stem Cells: A New Tool for Improving Tissue Regeneration

Margherita Maioli,*† Salvatore Rinaldi,‡§ Sara Santaniello,*† Alessandro Castagna,‡ Gianfranco Pigliaru,*† Sara Gualini,*† 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
§Medical School of Occupational Medicine, University of Florence, Florence, Italy
¶Cardiovascular Department, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy
#Bioscience Institute, Falciano, Republic of San Marino

Radiofrequency (RF) waves from Wi-Fi (wireless fidelity) technologies have become ubiquitous, with Internet access spreading into homes, and public areas. The human body harbors multipotent stem cells with various grading of potentiality. Whether stem cells may be affected by Wi-Fi RF energy remains unknown. We exposed mouse embryonic stem (ES) cells to a Radio Electric Asymmetric Conveyer (REAC), an innovative device delivering Wi-Fi RF of 2.4 GHz with its conveyer electrodes immersed into the culture medium. Cell responses were investigated by real-time PCR, Western blot, and confocal microscopy. Single RF burst duration, radiated power, electric and magnetic fields, specific absorption rate, and current density in culture medium were monitored. REAC stimulation primed transcription of genes involved in cardiac (GATA4, Nkx-2.5, and prodynorphin), skeletal muscle (myoD) and neuronal (neurogenin1) commitment, while downregulating the self renewal/pluripotency-associated genes Sox2, Oct4, and Nanog. REAC exposure enhanced the expression of cardiac, skeletal, and neuronal lineage-restricted marker proteins. The number of spontaneously beating ES-derived myocardial cells was also increased. In conclusion, REAC stimulation provided a “physical milieu” optimizing stem cell expression of pluripotentiality and the attainment of three major target lineages for regenerative medicine, without using chemical agonists or vector-mediated gene delivery.

Key words: Radiofrequency waves; Wifeless fidelity (Wi-Fi); Stem cells; Tissue regeneration

Received May 30, 2011; final acceptance July 31, 2011. Online prepub date: September 22, 2011.
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. 21, pp. 1135–1144, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627363
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bioreactor for Blood Product Production|

Michael R. Doran,* Ian Alexander Aird,† Flavia Marturana,† Nicholas Timmins,† Kerry Atkinson,‡ and Lars K. Nielsen†

*Stem Cell Therapies Laboratory, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia
†Bioengineering Laboratory, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Queensland, Australia
‡Adult Stem Cell Laboratory, Mater Medical Research Institute, Brisbane, Queensland, Australia

The feasibility of ex vivo blood production is limited by both biological and engineering challenges. From an engineering perspective, these challenges include the significant volumes required to generate even a single unit of a blood product, as well as the correspondingly high protein consumption required for such large volume cultures. Membrane bioreactors, such as hollow fiber bioreactors (HFBRs), enable cell densities approximately 100-fold greater than traditional culture systems and therefore may enable a significant reduction in culture working volumes. As cultured cells, and larger molecules, are retained within a fraction of the system volume, via a semipermeable membrane it may be possible to reduce protein consumption by limiting supplementation to only this fraction. Typically, HFBRs are complex perfusion systems having total volumes incompatible with bench scale screening and optimization of stem cell-based cultures. In this article we describe the use of a simplified HFBR system to assess the feasibility of this technology to produce blood products from umbilical cord blood-derived CD34+ hematopoietic stem progenitor cells (HSPCs). Unlike conventional HFBR systems used for protein manufacture, where cells are cultured in the extracapillary space, we have cultured cells in the intracapillary space, which is likely more compatible with the large-scale production of blood cell suspension cultures. Using this platform we direct HSPCs down the myeloid lineage, while targeting a 100-fold increase in cell density and the use of protein-free bulk medium. Our results demonstrate the potential of this system to deliver high cell densities, even in the absence of protein supplementation of the bulk medium.

Key words: Blood product; Bioreactor; Hematopoietic stem cell; Neutrophil (culture)

Received December 1, 2010; final acceptance June 30, 2011. Online prepub date: March 8, 2012.
Address correspondence to Dr. Michael R. Doran, B.Sc., B.Eng., Ph.D., Stem Cell Therapies Laboratory, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland Australia 4059. Tel: +61 (0)7 3138 0311; Fax: +61 (0)7 3138 6030; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1145–1160, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X600939
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Development and Application of Purified Tissue Dissociation Enzyme Mixtures for Human Hepatocyte Isolation

Roberto Gramignoli,*1 Michael L. Green,† Veysel Tahan,* Kenneth Dorko,* Kristen J. Skvorak,* Fabio Marongiu,‡ Wenchen Zao,§ Raman Venkataramanan,§ Ewa C. S. Ellis,¶ David Geller,# Andrew G. Breite,† Francis E. Dwulet,† Robert C. McCarthy,† and Stephen C. Strom*

*Department of Pathology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA
†VitaCyte LLC, Indianapolis, IN, USA
‡Department of Sciences and Biomedical Technologies, Experimental Pathology Section, University of Cagliari, Cagliari, Italy
§Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA, USA
¶Department of Clinical Science, Intervention and Technology (CLINTEC), Division of Transplantation Surgery, Liver Cell Lab, Karolinska University Hospital Huddinge, Stockholm, Sweden
#Department of Transplant Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA, USA

Human hepatocyte transplantation is gaining acceptance for the treatment of liver diseases. However, the reagents used to isolate hepatocytes from liver tissue are not standardized and show lot-to-lot variability in enzyme activity and endotoxin contamination. For clinical application, highly purified reagents are preferable to crude digest preparations. A purified tissue dissociating enzyme (TDE) preparation (CIzyme™ purified enzymes) was developed based on the enzyme compositions found in a superior lot of collagenase previously used by our group for human hepatocyte isolation. The performance of this enzyme preparation was compared to collagenase type XI on 110 liver cases by assessing hepatocyte yield, viability, and seven other functional assays that included plating efficiency, basal and induced CYP450 activities, phase II conjugation activity, and ammonia metabolism. No statistically significant difference was observed between these TDEs when they were used to isolate hepatocytes from liver resections or organ donor tissue on 54 hepatocyte isolations with type XI enzyme and 56 isolations using CIzyme™. These results show that a highly purified and defined TDE preparation can be formulated that provides excellent performance with respect to viability, yield, and functional activity of the isolated cells. In addition to reproducible formulation, these purified enzyme products have only 2–3% of the endotoxin of crude enzyme preparations. These results show that purified enzymes such as CIzyme™ will be a safe and effective for the isolation of human hepatocytes for clinical transplants.

Key words: Collagenase; Hepatocyte isolation; Human hepatocytes

Received September 30, 2010; final acceptance July 20, 2011. Online prepub date: November 11, 2011.
1Current address: Department of Laboratory Medicine, Karolinska Institutet and Hospital, Huddinge, Stockholm, Sweden.
Address correspondence to Stephen Strom, Ph.D., Professor, at his current address: Department of Laboratory Medicine, Karolinska Institutet and Hospital, Huddinge, SE 141 86, Stockholm, Sweden. E-mail: stephen.strom@ki.s


Cell Transplantation, Vol. 21, pp. 1261–1267, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X600984
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Remote Processing of Pancreas Can Restore Normal Glucose Homeostasis in Autologous Islet Transplantation After Traumatic Whipple Pancreatectomy: Technical Considerations

Aisha Khan,* Rahul M. Jindal,†‡§ Craig Shriver,§ Stephen R. Guy,¶ Amy E. Vertrees,§ Xiaojing Wang,* Xiumin Xu,* Joel Szust,* and Camillo Ricordi*

*Diabetes Research Institute and Cell Transplant Center, University of Miami, Miami, FL, USA
†Organ Transplant Service, Walter Reed Army Medical Center, Washington, DC, USA
‡Department of Medicine, George Washington University, Washington, DC, USA
§Department of Surgery, Walter Reed Army Medical Center, Washington, DC, USA
¶Drexel University, Philadelphia, PA, USA

An emergency autologous islet transplant after a traumatic Whipple operation and subsequent total pancreatectomy was performed for a 21-year-old patient who was wounded with multiple abdominal gunshot wounds. After Whipple pancreatectomy, the remnant pancreas (63.5 g), along with other damaged organs, was removed by the surgeons at Walter Reed Army Medical Center (WRAMC) and shipped to Diabetes Research Institute (DRI) for islet isolation. The pancreas was preserved in UW solution for 9.25 h prior to islet isolation. Upon arrival, the organ was visually inspected; the pancreatic head was missing, the rest of the pancreas was damaged and full of blood; the tail looked normal. A 16-gauge catheter was inserted into the main duct and directed towards tail of the pancreas after the dissection of main duct in the midbody of the pancreas. The pancreas was distended with collagenase solution (Roche MTF) through the catheter. During 10 min of intraductal delivery of enzyme, the gland was distended uniformly. No leakage of the solution was observed. The pancreas was transferred to a Ricordi chamber for automated mechanical and enzymatic digestion. Islets were purified using a COBE 2991 cell processor. Islet equivalents (IEQ; 221,250) of 40% purity and 90% viability were recovered during the isolation, which were shipped back to WRAMC and infused by intraportal injection into the patient. Immediate islet function was demonstrated by the rapid elevation of serum C peptide followed by insulin independence with near normal oral glucose tolerance test (OGTT) 1 and 2 months later. It is possible to restore near normal glucose tolerance with autologous islet transplantation after total pancreatectomy even with suboptimal number of islets while confirming that islets processed at a remote site are suitable for transplantation.

Key words: Autologous pancreatic islet cell transplant; Traumatic Whipple operation

Received November 10, 2010; final acceptance June 30, 2011. Online prepub date: September 22, 2011.
Address correspondence to Rahul M. Jindal, M.D., Ph.D., M.B.A., Organ Transplant Service, Walter Reed Army Medical Center, Washington, DC, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1269–1283, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627408
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Exendin-4 Increases the Expression of Hypoxia-Inducible Factor-1α in Rat Islets and Preserves the Endocrine Cell Volume of Both Free and Macroencapsulated Islet Grafts

Xiaohui Jia,*†1 Amit Sharma,*‡1 Makiko Kumagai-Braesch,* Annika M. Wernerson,§ Anne K. Sörenby,* Shinji Yamamoto,* Feng Wang,† and Annika B. Tibell*

*CLINTEC, Division of Transplantation Surgery, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden
†CLINTEC, Division of Surgery, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden
‡Division of Transplantation Surgery, Virginia Commonwealth University, Richmond, VA, USA
§CLINTEC, Division of Renal Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden

In this study, we evaluated the effects of exendin-4 on free and encapsulated islet grafts in a rodent model. We also investigated the role of a transcription factor, hypoxia-inducible factor-1 (HIF-1), in mediating the beneficial effects of exendin-4. Diabetic athymic mice were transplanted with free rat islets under the kidney capsule or with macroencapsulated rat islets SC with or without exendin-4, islet preculture (exendin-4 0.1 nM for 20 h), and/or recipient treatment (IP 100 ng/day, day 0–7). The mice were followed for 4 weeks and the graft function and β-cell volume were evaluated. The effects of exendin-4 on islet HIF-1α mRNA and protein expression and on ATP content in a rat insulinoma cell line (INS-1E) were also examined. Preculture with exendin-4 followed by recipient treatment improved the outcome of both free (73% graft function vs. 26% in controls, p = 0.03) and macroencapsulated islet grafts (100% vs. 25% in controls, p = 0.02). In macroencapsulated grafts, the exendin-4-treated group had significantly larger endocrine volume, less graft necrosis, and more blood vessels around the capsule. In rat islets cultured with exendin-4, HIF-1α mRNA and protein expression were significantly enhanced. ATP content was increased in exendin-4-treated INS-1E cells under hypoxic conditions. The improved functional outcome after transplantation of a marginal islet mass with a brief initial treatment with exendin-4 is related to a larger surviving endocrine cell volume. Exendin-4 may improve islet graft resistance to hypoxia during the peritransplant period by increasing the expression of HIF-1α.

Key words: Diabetes; Islet transplantation; Exendin-4; Macroencapsulation; Hypoxia-inducible factor-1α (HIF-1α)

Received September 27, 2010; final acceptance June 20, 2011. Online prepub date: March 8, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Makiko Kumagai-Braesch, CLINTEC, Division of Transplantation Surgery, F82, Karolinska University Hospital, Huddinge, S141 86, Stockholm, Sweden. Tel: +46-858582711; Fax: +46-858581390; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1285–1296, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X637371
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

LMP-420, a Small Molecular Inhibitor of TNF-α, Prolongs Islet Allograft Survival by Induction of Suppressor of Cytokine Signaling-1: Synergistic Effect With Cyclosporin-A

Nataraju Angaswamy,* Naohiko Fukami,* Venkataswarup Tiriveedhi,* George J. Cianciolo,† and T. Mohanakumar*‡

*Department of Surgery, Washington University School of Medicine, St. Louis, MO, USA
†Department of Pathology, Duke University Medical Center, Durham, NC, USA
‡Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA

Inflammatory insults following islet transplantation (ITx) hinders engraftment and long-term function of the transplanted (Tx) islets. Using a murine model of ITx, we determined the role of LMP-420, a novel TNF-α inhibitor, both individually and in combination with the immunosuppressant cyclosporine A (CSA) in islet engraftment and survival. Diabetic C57BL/6 mice were Tx with 500 BALB/c islets under the kidney capsule. Four cohorts were used: LMP-420 only, CSA only, combination of LMP-420 and CSA (LMP+CSA), and control (n = 12 per cohort). Serial monitoring of blood glucose levels revealed that LMP+CSA (35 ± 5 days) prolonged stable blood insulin levels compared to control (6 ± 4 days). Immunohistology demonstrated that coadministration (LMP+CSA) results in a significant decrease in CD8+ T-cell infiltration (LMP+CSA: 31 ± 18 vs. control: 224 ± 51 cells, p < 0.001). Serum cytokine analysis revealed that LMP-420 administration resulted in an increase in the anti-inflammatory cytokine IL-10 (2.5-fold), and a decrease in TNF-α  (threefold) with no change in IL-2. However, coadministration resulted in a marked decrease in both IL-2 and TNF-α  (threefold) along with increase in IL-10 (threefold). Coadministration also demonstrated increase of antiapoptotic SOCS-1 and Mn-SOD expression and significant reduction of donor-specific antibodies (p < 0.005). In conclusion, LMP-420 administration with CSA results in the upregulation of anti-inflammatory and antiapoptotic mechanisms which facilitate islet allograft engraftment and survival.

Key words: LMP-420; Cyclosporin A; Islet transplantation; Type 1 diabetes mellitus; Tumor necrosis factor-α  (TNF-α)

Received Feburary 25, 2011; final acceptance July 5, 2011. Online prepub date: March 16, 2012.
Address correspondence to T. Mohanakumar, Ph.D., Department of Surgery, Washington University School of Medicine, Box 8109, 3328 CSRB, 660 S. Euclid Ave, St. Louis, MO 63110, USA. Tel: (314) 362-8463; Fax: (314) 747-1560; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1197–1304, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X 603620
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Metabolic Function of a Suboptimal Transplanted Islet Mass in Nonhuman Primates on Rapamycin Monotherapy

Scott A. Soleimanpour,*† Boaz Hirshberg,* David J. Bunnell,* Anne E. Sumner,‡ Marilyn Ader,§ Alan T. Remaley,¶ Kristina I. Rother,‡ Michael R. Rickels,† and David M. Harlan*#

*Islet and Autoimmunity Branch, NIDDK, NIH, Bethesda, MD, USA
†Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
‡Diabetes, Endocrinology, and Obesity Branch, NIDDK, NIH, Bethesda, MD, USA
§Department of Physiology and Biophysics, University of Southern California, Los Angeles, CA, USA
¶Department of Laboratory Medicine, NIH, Bethesda, MD, USA
#Diabetes Division, Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA

Although islet transplantation may restore insulin independence to individuals with type 1 diabetes mellitus, most have abnormal glucose tolerance. We asked whether the defective glucose tolerance is due to inadequate β-cell mass or to impaired insulin sensitivity. We performed metabolic studies on four cynomolgus primates before inducing diabetes with streptozotocin (STZ), then again 2–3 weeks after restoring insulin independence via intrahepatic islet transplantation utilizing a calcineurin inhibitor-free immunosuppressive regimen (induction with rabbit antithymocyte globulin and maintenance therapy with rapamycin). Engrafted β-cell mass was assessed by acute insulin and C-peptide responses to glucose (AIRglu and ACRglu) and arginine (AIRarg and ACRarg). Insulin sensitivity (SI) was determined in naive and transplanted primates from an intravenous glucose tolerance test using the minimal model. α-Cell function was determined by the acute glucagon response to arginine (AGRarg). Glucose tolerance (Kg) decreased from 4.1 ± 0.5%/min in naive primates to 1.8 ± 0.3%/min in transplanted primates (p < 0.01). Following transplantation, AIRglu was 28.7 ± 13.1 μU/ml compared to 169.9 ± 43.1 μU/ml (p < 0.03) in the naive condition, ACRglu was 14.5 ± 6.0 ng/ml compared to 96.5 ± 17.0 ng/ml naive (p < 0.01), AIRarg was 29.1 ± 13.1 μU/ml compared to 91.4 ± 28.2 μU/ml naive (p < 0.05), and ACRarg was 1.11 ± 0.51 ng/ml compared to 2.79 ± 0.77 ng/ml naive (p < 0.05). SI did not differ from naive to posttransplant states. AGRarg was reduced in transplanted primates (349 ± 118 pg/ml) when compared to both naive (827 ± 354 pg/ml) and post-STZ diabetic primates (1020 ± 440 pg/ml) (p < 0.01 for both comparisons). These data suggest that impaired glucose tolerance observed in islet transplant recipients is secondary to low functional β-cell mass and not to insulin resistance shortly after transplant. Furthermore, improved glycemic control achieved via islet transplantation over the diabetic state might be attained, in part, via reduced glucagon secretion.

Key words: Nonhuman primates; Islet transplantation; Metabolic function

Received March 4, 2011; final acceptance June 15, 2011. Online prepub date: November 11, 2011.
Address correspondence to Scott A. Soleimanpour, M.D., Instructor of Medicine, Institute for Diabetes, Obesity, and Metabolism, University of Pennsylvania School of Medicine, 3400 Civic Center Blvd., Bldg. 421, Translational Research Center, Room 12-165, Philadelphia, PA 19104, USA. Tel: (215) 898-0209; Fax: (215) 898-5408; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1305–1320, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X 627534
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Microvesicles Derived From Endothelial Progenitor Cells Enhance Neoangiogenesis of Human Pancreatic Islets

Vincenzo Cantaluppi,*†1 Luigi Biancone,*†1 Federico Figliolini,* Silvia Beltramo,* Davide Medica,* Maria Chiara Deregibus,* Francesco Galimi,‡ Renato Romagnoli,§ Mauro Salizzoni,§ Ciro Tetta,¶ Giuseppe Paolo Segoloni,† and Giovanni Camussi*†

*Research Center for Experimental Medicine (CeRMS), University of Torino, Torino, Italy
†Nephrology, Dialysis and Kidney Transplantation Center, Department of Internal Medicine, University of Torino, Torino, Italy
‡Department of Biomedical Sciences/INBB, University of Sassari, Sassari, Italy
§Liver Transplantation Center, University of Torino, Torino, Italy
¶SisTER, Palazzo Pignano, Italy and Fresenius Medical Care, Bad Homburg, Germany

The efficacy of islet transplantation is limited by poor graft vascularization. We herein demonstrated that microvesicles (MVs) released from endothelial progenitor cells (EPCs) enhanced human islet vascularization. After incorporation into islet endothelium and β-cells, EPC-derived MVs favored insulin secretion, survival, and revascularization of islets transplanted in SCID mice. MVs induced in vitro islet endothelial cell proliferation, migration, resistance to apoptosis, and organization in vessel-like structures. Moreover, MVs partially overcame the antiangiogenic effect of rapamycin and inhibited endothelial–leukocyte interaction via L-selectin and CD40. MVs were previously shown to contain defined patterns of mRNAs. Here we demonstrated that MVs carried the proangiogenic miR-126 and miR-296 microRNAs (miRNAs). MVs pretreated with RNase or derived from Dicer knocked-down EPCs showed a reduced angiogenic effect. In addition, MVs overcame the antiangiogenic effect of the specific antagomiRs of miR-126 and miR-296, suggesting a relevant contribution of miRNAs delivered by MVs to islet endothelium. Microarray analysis of MV-stimulated islet endothelium indicated the upregulation of mRNAs coding for factors involved in endothelial proliferation, differentiation, and angiogenesis. In addition, MVs induced the activation of the PI3K-Akt and eNOS signaling pathways in islet endothelium. These results suggest that MVs activate an angiogenic program in islet endothelium that may sustain revascularization and β-cell function.

Key words: Microvesicles; Exosomes; Endothelial progenitors; Islet transplantation; Angiogenesis; MicroRNAs

Received December 22, 2010; final acceptance July 7, 2011. Online prepub date: March 22, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Prof. Giovanni Camussi, Cattedra di Nefrologia, Dipartimento di Medicina Interna, Ospedale Maggiore S. Giovanni Battista “Molinette,” Corso Dogliotti 14, 10126, Torino, Italy. Tel: +39-011-6336708; Fax: +39-011-6631184; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1321–1339, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X 612530
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Generation of Glucose-Responsive, Insulin-Producing Cells From Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells

Kamalaveni R. Prabakar,* Juan Domínguez-Bendala,*† R. Damaris Molano,* Antonello Pileggi,*† Susana Villate,* Camillo Ricordi,*† and Luca Inverardi*‡§

*Diabetes Research Institute, University of Miami Miller School of Medicine, Miami, FL, USA
†Department of Surgery, University of Miami Miller School of Medicine, Miami, FL, USA
‡Department of Medicine, University of Miami Miller School of Medicine, Miami, FL, USA
§Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, FL, USA

We sought to assess the potential of human cord blood-derived mesenchymal stem cells (CB-MSCs) to derive insulin-producing, glucose-responsive cells. We show here that differentiation protocols based on stepwise culture conditions initially described for human embryonic stem cells (hESCs) lead to differentiation of cord blood-derived precursors towards a pancreatic endocrine phenotype, as assessed by marker expression and in vitro glucose-regulated insulin secretion. Transplantation of these cells in immune-deficient animals shows human C-peptide production in response to a glucose challenge. These data suggest that human cord blood may be a promising source for regenerative medicine approaches for the treatment of diabetes mellitus.

Key words: Umbilical cord blood; Islets of Langerhans; Pancreas; Adult stem cells; Mesenchymal stem cells (MSCs)

Received December 2, 2010; final acceptance June 22, 2011. Online prepub date: December 21, 2011.
Address correspondence to Camillo Ricordi, M.D., Director, Diabetes Research Institute and Cell Transplant Center, University of Miami Miller School of Medicine, 1450 NW 10th Avenue, Miami, FL 33136, USA. Tel: 305 243-6913; Fax: 305 243-4404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it