Cell Transplantation 22(12) Abstracts

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Cell Transplantation, Vol. 22, pp. 2187-2201, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657882
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Interferon-β Delivery Via Human Neural Stem Cell Abates Glial Scar Formation in Spinal Cord Injury

Yusuke Nishimura,* Atsushi Natsume,* Motokazu Ito,* Masahito Hara,* Kazuya Motomura,* Ryuichi Fukuyama,† Naoyuki Sumiyoshi,† Ichio Aoki,‡ Tsuneo Saga,‡ Hong J. Lee,§ Toshihiko Wakabayashi,* and Seung U. Kim§¶

*Department of Neurosurgery, Nagoya University, Nagoya, Japan
†Division of Pathology, Konan Kosei Hospital, Aichi, Japan
‡MR Molecular Imaging Team, Molecular Imaging Center, National Institute of Radiological Sciences, Chiba, Japan
§Medical Research Institute, Chung-Ang University College of Medicine, Seoul, Korea
¶Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, Canada

Glial scar formation is the major impedance to axonal regrowth after spinal cord injury (SCI), and scar-modulating treatments have become a leading therapeutic goal for SCI treatment. In this study, human neural stem cells (NSCs) encoding interferon-β (INF-β) gene were administered intravenously to mice 1 week after SCI. Animals receiving NSCs encoding IFN-β exhibited significant neurobehavioral improvement, electrophysiological recovery, suppressed glial scar formation, and preservation of nerve fibers in lesioned spinal cord. Systemic evaluation of SCI gliosis lesion site with lesion-specific microdissection, genome-wide microarray, and MetaCore pathway analysis identified upregulation of toll-like receptor 4 (TLR4) in SCI gliosis lesion site, and this led us to focus on TLR4 signaling in reactive astrocytes. Examination of primary astrocytes from TLR4 knockout mice, and in vivo inhibition of TLR4, revealed that the effect of IFN-β on the suppression of glial scar formation in SCI requires TLR4 stimulation. These results suggest that IFN-β delivery via intravenous injection of NSCs following SCI inhibits glial scar formation in spinal cord through stimulation of TLR4 signaling.

Key words: Human neural stem cells (NSCs); Interferon-β (INF-β) gene; Spinal cord injury (SCI); Toll-like receptor 4 (TLR4)

Received May 31, 2012; final acceptance September 30, 2012. Online prepub date: October 12, 2012.
Address correspondence to Atsushi Natsume, M.D., Ph.D., Department of Neurosurgery, Nagoya University School of Medicine, Nagoya 466-8550, Japan. Tel: +81-52-744-2353; Fax: +81-52-744-2360; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Seung U. Kim, M.D., Ph.D., Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia Vancouver, BC V6T 2B5, Canada. Tel: +82-2-820-5652; Fax: +82-2-813-5387; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2203-2217, 2013
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DOI: http://dx.doi.org/10.3727/096368912X658872
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Combining Neurotrophin-Transduced Schwann Cells and Rolipram to Promote Functional Recovery From Subacute Spinal Cord Injury

Govinder Flora,*†1 Gravil Joseph,*†1 Samik Patel,*† Amanpreet Singh,*† Drew Bleicher,*† David J. Barakat,*† Jack Louro,*† Stephanie Fenton,*† Maneesh Garg,*† Mary Bartlett Bunge,*†‡§¶ and Damien D. Pearse*†‡¶

*The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, USA
†The Neuroscience Program, University of Miami Miller School of Medicine, Miami, FL, USA
‡Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, USA
§Department of Cell Biology, University of Miami Miller School of Medicine, Miami, FL, USA
¶The Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL, USA

Following spinal cord injury (SCI), both an inhibitory environment and lack of intrinsic growth capacity impede axonal regeneration. In a previous study, prevention of cyclic adenosine monophosphate (AMP) hydrolysis by the phosphodiesterase-4 inhibitor rolipram, in combination with Schwann cell (SC) grafts, promoted significant supraspinal and proprioceptive fiber growth and/or sparing and improved locomotion. In another study, transplanted SCs transduced to generate a bifunctional neurotrophin (D15A) led to significant increases in graft SCs and axons, including supraspinal and myelinated axons. Here we studied the growth and myelination of local and supraspinal axons and functional outcome following the combination of rolipram administration and neurotrophin-transduced SC implantation after SCI. Rolipram was administered subcutaneously for 4 weeks immediately after contusion at vertebral T8 (25.0-mm weight drop, MASCIS impactor). GFP or GFP-D15Atransduced SCs were injected into the injury epicenter 1 week after SCI. GFP-D15A SC grafts and GFP SC grafts with rolipram contained significantly more serotonergic fibers compared to GFP SCs. SC myelinated axons were increased significantly in GFP SC with rolipram-treated animals compared to animals receiving SCI alone. Rolipram administered with either GFP or GFP-D15A SCs significantly increased numbers of brain stem-derived axons below the lesion/implant area and improved hindlimb function. Compared to the single treatments, the combination led to the largest SC grafts, the highest numbers of serotonergic fibers in the grafts, and increased numbers of axons from the reticular formation below the lesion/implant area and provided the greatest improvement in hindlimb function. These findings demonstrate the therapeutic potential for a combination therapy involving the maintenance of cyclic AMP levels and neurotrophin-transduced SCs to repair the subacutely injured spinal cord.

Key words: Cyclic adenosine monophosphate (AMP); Viral vector; Supraspinal axons; Proprioceptive fiber growth; Myelination; Brain-derived neurotrophic factor (BDNF); Neurotrophin-3 (NT-3); Axonal growth

Received March 8, 2012; final acceptance October 30, 2012. Online prepub date: November 8, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Damien D. Pearse, Ph.D., Lois Pope LIFE Center, P.O. Box 016960, Mail locator R-48, Miami, FL 33101, USA. Tel: +1-(305)-243-7139; Fax: +1-(305)-898-3777; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2219-2236, 2013
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DOI: http://dx.doi.org/10.3727/096368912X661373
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Cotransplantation of Glial Restricted Precursor Cells and Schwann Cells Promotes Functional Recovery After Spinal Cord Injury

Jian-Guo Hu,*†1 Xiao-Fei Wang,*1 Ling-Xiao Deng,* Nai-Kui Liu,* Xiang Gao,* Jing-Hui Chen,* Feng-Cheng Zhou,‡ and Xiao-Ming Xu*‡

*Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery, Indiana University School of Medicine, Indianapolis, IN, USA
†Anhui Key Laboratory of Tissue Transplantation, The First Affiliated Hospital, Bengbu Medical College, Bengbu, P.R. China
‡Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN, USA

Oligodendrocyte (OL) replacement can be a promising strategy for spinal cord injury (SCI) repair. However, the poor posttransplantation survival and inhibitory properties to axonal regeneration are two major challenges that limit their use as donor cells for repair of CNS injuries. Therefore, strategies aimed at enhancing the survival of grafted oligodendrocytes as well as reducing their inhibitory properties, such as the use of more permissive oligodendrocyte progenitor cells (OPCs), also called glial restricted precursor cells (GRPs), should be highly prioritized. Schwann cell (SC) transplantation is a promising translational strategy to promote axonal regeneration after CNS injuries, partly due to their expression and secretion of multiple growth-promoting factors. Whether grafted SCs have any effect on the biological properties of grafted GRPs remains unclear. Here we report that either SCs or SC-conditioned medium (SCM) promoted the survival, proliferation, and migration of GRPs in vitro. When GRPs and SCs were cografted into the normal or injured spinal cord, robust survival, proliferation, and migration of grafted GRPs were observed. Importantly, grafted GRPs differentiated into mature oligodendrocytes and formed new myelin on axons caudal to the injury. Finally, cografts of GRPs and SCs promoted recovery of function following SCI. We conclude that cotransplantation of GRPs and SCs, the only two kinds of myelin-forming cells in the nervous system, act complementarily and synergistically to promote greater anatomical and functional recovery after SCI than when either cell type is used alone.

Key words: Glial restricted precursor cells (GRPs); Schwann cells (SCs); Transplantation; Spinal cord injury (SCI); Functional recovery

Received August 5, 2011; final acceptance July 30, 2012. Online prepub date: January 2, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Xiao-Ming Xu, Ph.D., Professor and Scientific Director, Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Indiana University School of Medicine, 950 W. Walnut Street, R-II Bldg, Room 402, Indianapolis, IN 46202, USA. Tel: +1-317-274-1036; Fax: +1-317-278-5849; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2237-2256, 2013
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DOI: http://dx.doi.org/10.3727/096368912X658809
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Transplantation of GABAergic Cells Derived From Bioreactor-Expanded Human Neural Precursor Cells Restores Motor and Cognitive Behavioral Deficits in a Rodent Model of Huntington’s Disease

Marcus C. McLeod,*1 Nao R. Kobayashi,*1,2 Arindom Sen,† Behnam A. Baghbaderani,† Damaso Sadi,* Ruperto Ulalia,* Leo A. Behie,† and Ivar Mendez*

*Cell Restoration Laboratory, Brain Repair Centre, Dalhousie University, Halifax, Nova Scotia, Canada
†Pharmaceutical Production Research Facility (PPRF), Schulich School of Engineering, University of Calgary, Calgary, Alberta, Canada

Huntington’s disease (HD) is a neurodegenerative disorder that is characterized by progressive dementia, choreiform involuntary movements, and emotional deterioration. Neuropathological features include the progressive degeneration of striatal g-aminobutyric acid (GABA) neurons. New therapeutic approaches, such as the transplantation of human neural precursor cells (hNPCs) to replace damaged or degenerated cells, are currently being investigated. The aim of this study was to investigate the potential for utilizing telencephalic hNPCs expanded in suspension bioreactors for cell restorative therapy in a rodent model of HD. hNPCs were expanded in a hydrodynamically controlled and homogeneous environment under serum-free conditions. In vitro analysis revealed that the bioreactor-expanded telencephalic (BET)-hNPCs could be differentiated into a highly enriched population of GABAergic neurons. Behavioral assessments of unilateral striatal quinolinic acid-lesioned rodents revealed a significant improvement in motor and memory deficits following transplantation with GABAergic cells differentiated from BET-hNPCs. Immunohistochemical analysis revealed that transplanted BET-hNPCs retained a GABAergic neuronal phenotype without aberrant transdifferentiation or tumor formation, indicating that BET-hNPCs are a safe source of cells for transplantation. This preclinical study has important implications as the transplantation of GABAergic cells derived from predifferentiated BET-hNPCs may be a safe and feasible cell replacement strategy to promote behavioral recovery in HD.

Key words: Huntington’s disease (HD); Human neural stem/precursor cells; Suspension bioreactor; Neural cell transplantation; g-Aminobutyric acid (GABA); Behavioral recovery

Received February 27, 2011; final acceptance October 13, 2012. Online prepub date: November 1, 2012.
1These authors provided equal contribution to this work.
2Current address: Centre for Neural Engineering, The University of Melbourne, Victoria, 3010, Australia.
Address correspondence to Dr. Ivar Mendez, M.D., Ph.D., at his current address: University of Saskatchewan and Saskatoon Health Region, Department of Surgery and Royal University Hospital, Suite 2646, 103 Hospital Drive, Saskatoon, SK S7N 0W8, Canada. Tel: (306) 966-8641; Fax: (306) 966-8026; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2257-2266, 2013
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DOI: http://dx.doi.org/10.3727/096368912X662435
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Targeted Cell Reprogramming Produces Analgesic Chromaffin-Like Cells From Human Mesenchymal Stem Cells

T. Qu,* G. Shi,* K. Ma,* H. N. Yang,† W. M. Duan,‡ and G. D. Pappas*

*Department of Psychiatry, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA
†Department of Neural Medicine, The Second Hospital of Shandong University, Jinan, Shandong, China
‡Department of Anatomy, Capital Medical University, Youanmenwai, Fengtai District, Beijing, China

Transplantation of allogeneic adrenal chromaffin cells demonstrated the promise of favorable outcomes for pain relief in patients. However, there is a very limited availability of suitable human adrenal gland tissues, genetically well-matched donors in particular, to serve as grafts. Xenogeneic materials, such as porcine and bovine adrenal chromaffin cells, present problems; for instance, immune rejection and possible pathogenic contamination are potential issues. To overcome these challenges, we have tested the novel approach of cell reprogramming to reprogram human bone marrow (BM)-derived mesenchymal stem cells (hMSCs) using cellular extracts of porcine chromaffin cells. We produced a new type of cell, chromaffin-like cells, generated from the reprogrammed hMSCs, which displayed a significant increase in expression of human preproenkephalin (hPPE), a precursor for enkephalin opioid peptides, compared to the inherent expression of hPPE in naive hMSCs. The resultant chromaffin-like cells not only expressed the key molecular markers of adrenal chromaffin cells, such as tyrosine hydroxylase (TH) and methionine enkephalin (Met-enkephalin), but also secreted opioid peptide Met-enkephalin in culture. In addition, intrathecal injection of chromaffin-like cells in rats produced significant analgesic effects without using immunosuppressants. These results suggest that analgesic chromaffin-like cells can be produced from an individual’s own tissue-derived stem cells by targeted cell reprogramming and also that these chromaffin-like cells may serve as potential autografts for chronic pain management.

Key words: Mesenchymal stem cells; Adrenal chromaffin cells; Cell reprogramming; Pain management; Autologous stem cells

Received March 29, 2012; final acceptance November 10, 2012. Online prepub date: February 4, 2013.
Address correspondence to Tingyu Qu, M.D., Ph.D., Department of Psychiatry, College of Medicine, University of Illinois at Chicago (UIC), 1601 West Taylor Street, Chicago, IL 60612, USA. Tel: +1-(312) 355-1786; Fax: +1-(312) 413-1177; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2267-2277, 2013
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DOI: http://dx.doi.org/10.3727/096368911X582769
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Allogeneic Mesenchymal Stem Cell Transplantation in Severe and Refractory Systemic Lupus Erythematosus: 4 Years of Experience

Dandan Wang,* Huayong Zhang,* Jun Liang,* Xia Li,* Xuebing Feng,* Hong Wang,* Bingzhu Hua,* Bujun Liu,* Liwei Lu,† Gary S. Gilkeson,‡ Richard M. Silver,‡ Wanjun Chen,§ Songtao Shi,¶ and Lingyun Sun*

*Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, Jiangsu, P. R. China
†Department of Pathology and Microbiology, Center of Infection and Immunology, Hong Kong University, Hong Kong, China
‡Division of Rheumatology, Medical University of South Carolina, Charleston, SC, USA
§Mucosal Immunology Unit, Oral Infection and Immunity Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD, USA
¶Center for Craniofacial Molecular Biology, University of Sothern California School of Dentistry, Los Angeles, CA, USA

Mesenchymal stem cells (MSCs) are multipotential nonhematopoietic progenitors and are capable of differentiating into several tissues of mesenchymal origin. We have shown that bone marrow-derived MSCs from both SLE patients and lupus-prone MRL/lpr mice are defective structurally and functionally. Here we observe the long-term safety and efficacy of allogeneic MSC transplantation (MSCT) in treatment-resistant SLE patients. Eighty-seven patients with persistently active SLE who were refractory to standard treatment or had life-threatening visceral involvement were enrolled. Allogeneic bone marrow or umbilical cord-derived MSCs were harvested and infused intravenously (1 × 106 cells/kg of body weight). Primary outcomes were rates of survival, disease remission and relapse, as well as transplantation-related adverse events. Secondary outcomes included SLE disease activity index (SLEDAI) and serologic features. During the 4-year follow-up and with a mean follow-up period of 27 months, the overall rate of survival was 94% (82/87). Complete clinical remission rate was 28% at 1 year (23/83), 31% at 2 years (12/39), 42% at 3 years (5/12), and 50% at 4 years (3/6). Rates of relapse were 12% (10/83) at 1 year, 18% (7/39) at 2 years, 17% (2/12) at 3 years, and 17% (1/6) at 4 years. The overall rate of relapse was 23% (20/87). Disease activity declined as revealed by significant changes in the SLEDAI score, levels of serum autoantibodies, albumin, and complements. A total of five patients (6%) died after MSCT from non-treatment-related events in the 4-year follow-up, and no transplantation-related adverse event was observed. Allogeneic MSCT resulted in the induction of clinical remission and improvement in organ dysfunction in drug-resistant SLE patients.

Key words: Mesenchymal stem cells (MSCs); Lupus erythematosus; Transplantation; Clinical remission

Received April 14, 2012; final acceptance October 16, 2012. Online prepub date: October 31, 2012.
Address correspondence to Lingyun Sun, Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, P. R. China. Tel: +86-25-8310-5219; Fax: +86-25-8310-5209; 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. 2279-2290, 2013
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DOI: http://dx.doi.org/10.3727/096368912X658692
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Allogenic Mesenchymal Stem Cell Transplantation Ameliorates Nephritis in Lupus Mice Via Inhibition of B-Cell Activation

Xiaolei Ma,* Nan Che,* Zhifeng Gu,* Jing Huang,* Dandan Wang,* Jun Liang,* Yayi Hou,* Gary Gilkeson,† Liwei Lu, ‡ and Lingyun Sun*

*Department of Immunology and Rheumatology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, China
†Department of Rheumatology, Medical University of South Carolina, Charleston, SC, USA
‡Department of Pathology and Center of Infection and Immunology, The University of Hong Kong, Hong Kong, China

Recent evidence indicates that bone marrow-derived mesenchymal stem cells (BM-MSCs) possess immunosuppressive properties both in vitro and in vivo. We have previously demonstrated that transplantation of human MSCs can significantly improve the autoimmune conditions in MRL/lpr mice. The current study aimed to determine the mechanisms by which murine BM-MSC transplantation (MSCT) ameliorates nephritis in MRL/lpr mice. In this study, we found that MSCT can significantly prolong the survival of MRL/lpr mice. Eight weeks after transplantation, MSCT-treated mice showed significantly smaller spleens than control animals, with fewer marginal zones (MZs), T1, T2, activated B-cells, and plasma cells. Moreover, serum levels of B-cell activating factor (BAFF) and IL-10 in MSCT-treated mice decreased significantly compared to those in the control group, while levels of serum TGF-β were increased. Notably, decreased BAFF expression in both spleen and kidney was accompanied by decreased production of anti-dsDNA autoantibodies and proteinuria in MSCT-treated mice. Since BAFF is mainly expressed by T-cells and dendritic cells, we incubated BM-MSCs and DCs together and found that the production of BAFF by DCs was suppressed by MSCs. Thus, our findings suggest that MSCT may suppress the excessive activation of B-cells via inhibition of BAFF production in MRL/lpr mice.

Key words: Systemic lupus erythematosus; Mesenchymal stem cells; Transplantation; B-cell activating factor; B lymphocyte

Received February 20, 2012; final acceptance October 16, 2012. Online prepub date: October 31, 2012.
Address correspondence to Lingyun Sun, M.D., Ph.D., Department of Immunology and Rheumatology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, Jiangsu 210008, P. R. China. 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. 2291-2298, 2013
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DOI: http://dx.doi.org/10.3727/096368912X658818
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Feasibility of Delivering Mesenchymal Stem Cells Via Catheter to the Proximal End of the Lesion Artery in Patients With Stroke in the Territory of the Middle Cerebral Artery

Yongjun Jiang,1 Wusheng Zhu,1 Juehua Zhu, Li Wu, Gelin Xu, and Xinfeng Liu

Department of Neurology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China

Stem cell-based therapy shows great potential in stroke patients. Intra-artery infusion exhibits greater biological distribution compared to intravenous delivery. In addition, umbilical cord mesenchymal stem cells (UCMSCs) have several advantages compared with other types of stem cells. The aim of this study was to evaluate the safety and efficacy of UCMSCs delivered by a catheter to a near lesion site for treatment of an infarction in the middle cerebral artery territory. Four patients with stroke (three with ischemic and one with hemorrhagic stroke) in the middle cerebral artery territory were recruited in this study. One single dose of 2 × 107 UCMSCs was infused within 20 min via catheterization in the M1 segment of the middle cerebral artery. The safety and efficacy of this approach were assessed during the in-hospital and 6-month follow-up evaluation. The cell delivery was successfully performed in all of the patients, and no major accidents (stroke or death) were observed. Moreover, no fever or rash was reported. After cellular therapy, two of the three ischemic stroke patients demonstrated improved muscle strength. The improvement of the modified Rankin scale was observed in two patients, both of whom suffered from ischemic stroke at 90 and 180 days after the stem cell therapy. The hemorrhagic stroke patient failed to demonstrate improved muscle strength and did not amend his daily activities. Intra-artery delivery of UCMSCs via catheterization was a feasible and safe approach and may improve the neurological function of ischemic stroke patients with the middle cerebral artery territory infarcts.

Key words: Stem cells; Stroke; Middle cerebral artery; Intra-artery

Received June 18, 2012; final acceptance October 16, 2012. Online prepub date: November 1, 2012.
1These authors provided equal contribution to this study.
Address correspondence to Xinfeng Liu or Gelin Xu, Department of Neurology, Jinling Hospital, Nanjing University School of Medicine, 305 Zhongshan East Road, Nanjing 21002, China. Tel: +86 25 8527 2631; Fax +86 25 8480 5169; 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. 2299-2309, 2013
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DOI: http://dx.doi.org/10.3727/096368912X658016
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Catheter-Based Endomyocardial Delivery of Mesenchymal Precursor Cells Using 3D Echo Guidance Improves Cardiac Function in a Chronic Myocardial Injury Ovine Model

Yanping Cheng,* Genghua Yi,* Gerard B. Conditt,* Alexander Sheehy,† Frank D. Kolodgie,‡ Armando Tellez,* Igor Polyakov,† Anguo Gu,* Michael S. Aboodi,* David Wallace-Bradley,* Michael Schuster,§ Timothy Martens,§ Silviu Itescu,§ Greg L. Kaluza,* Shubhayu Basu,† Renu Virmani,‡ Juan F. Granada,* and Warren Sherman*†

*The Jack Skirball Center for Cardiovascular Research, Cardiovascular Research Foundation, Orangeburg, NY, USA
†Abbott Vascular, Santa Clara, CA, USA
‡CVPath Institute, Inc., Gaithersburg, MD, USA
§Angioblast Systems, Inc., New York, NY, USA
†Columbia University College of Physicians and Surgeons, Columbia University, New York, NY, USA

The administration of bone marrow-derived stem cells may provide a new treatment option for patients with heart failure. Transcatheter cell injection may require multi-imaging modalities to optimize delivery. This study sought to evaluate whether endomyocardial injection of mesenchymal precursor cells (MPCs) could be guided by real-time 3D echocardiography (RT3DE) in treating chronic, postinfarction (MI) left ventricular (LV) dysfunction in sheep. Four weeks after induction of an anterior wall myocardial infarction in 39 sheep, allogeneic MPCs in doses of either 25 × 106 (n = 10), 75 × 106 (n = 9), or 225 × 106 (n = 10) cells or nonconditioned control media (n = 10) were administered intramyocardially into infarct and border zone areas using a catheter designed for combined fluoroscopic and RT3DE-guided injections. LV function was assessed before and after injection. Infarct dimension and vascular density were evaluated histologically. RT3DE-guided injection procedures were safe. Compared to controls, the highest dose MPC treatment led to increments in ejection fraction (3 ± 3% in 225M MPCs vs. –5 ± 4% in the control group, p < 0.01) and wall thickening in both infarct (4 ± 4% in 225M MPCs vs. −3 ± 6% in the control group, p = 0.02) and border zones (4 ± 6% in 225M MPCs vs. −8 ± 9% in the control group, p = 0.01). Histology analysis demonstrated significantly higher arteriole density in the infarct and border zones in the highest dose MPC-treated animals compared to the lower dose or control groups. Endomyocardial implantation of MPCs under RT3DE guidance was safe and without observed logistical obstacles. Significant increases in LV performance (ejection fraction and wall thickening) and neovascularization resulted from this technique, and so this technique has important implications for treating patients with postischemic LV dysfunction.

Key words: Stem cells; Catheters; Echocardiography; Heart failure; Myocardial infarction (MI)

Received April 16, 2012; final acceptance September 30, 2012. Online prepub date: November 25, 2012.
Address correspondence to Warren Sherman, M.D., Columbia University Medical Center, 161 Fort Washington, New York, NY 10032, USA. Tel: +1 212 343 3612; Fax: +1 646 619 4329; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2311-2321, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657981
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Donor-Matched Functional and Molecular Characterization of Canine Mesenchymal Stem Cells Derived From Different Origins

Sun-A Ock,*† Geun-ho Maeng,* Yeon-Mi Lee,* Tae-Ho Kim,* B. Mohana Kumar,*‡ Sung-Lim Lee,*‡ and Gyu-Jin Rho*‡

*Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Jinju, Republic of Korea
†Animal Biotechnology Division, National Institute of Animal Science, RDA, Kwonsun-Gu, Suwon, Republic of Korea
‡Research Institute of Life Sciences, Gyeongsang National University, Jinju, Republic of Korea

Canine mesenchymal stem cells (cMSCs) have generated a great interest as a promising source for cell-based therapies. To understand the basic biological properties of cMSCs derived from bone marrow (cBM-MSCs), adipose tissue (cA-MSCs), and dermal skin (cDS-MSCs) from a single donor, the present study compared their alkaline phosphatase (AP) activity, expression of CD markers and stem cell transcription factors, differentiation ability into osteogenic, adipogenic, and chondrogenic lineages, in vivo ectopic bone formation, chromosomal stability, cell cycle status, telomere length, and telomerase activity. Expressions of AP activity and transcription factors (Oct3/4, Nanog, and Sox2) were either absent or extremely weak in all cMSCs. CD marker profile (CD45, CD90+, and CD105+) and differentiation capacity were exhibited by all cMSCs, although cAMSCs had enhanced cytochemical staining associated with expression of lineage-specific markers. In vivo bone formation of cMSCs was performed with demineralized bone matrix (DBM) by transplanting into the subcutaneous spaces of 9-week-old BALB/c-nu mice, followed by radiographic and histological analysis after 1 and 2 months. cA-MSCs and cDS-MSCs, in contrast to the in vitro observations, also displayed higher in vivo osteogenic abilities than cBM-MSCs. Ploidy analysis showed that cells were diploid and contained no noticeable chromosomal abnormalities. Furthermore, a relatively low percentage of cells was found at the G1 phase in all cMSCs, especially in DS-MSCs. Regardless of the different tissue sources, cMSCs from a single donor showed no differences in telomere lengths (~18–19 kbp) but exhibited varied telomerase activity. The above results suggest that tissue-specific cMSCs derived from a single donor possess slight differences in stem cell properties.

Key words: Canine; Mesenchymal stem cells; Transcription factors; Telomere length; Telomerase reverse transcriptase (TERT) expression; In vivo ectopic bone formation

Received July 27, 2011; final acceptance October 9, 2012. Online prepub date: October 12, 2012.
Address correspondence to Gyu-Jin Rho, D.V.M., Ph.D., Department of Theriogenology and Biotechnology, College of Veterinary Medicine/Research Institute of Life Sciences, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea. Tel: +82-55-772-2347; Fax: +82-55-772-2349; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2323-2333, 2013
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DOI: http://dx.doi.org/10.3727/096368912X662417
E-ISSN 1555-3892
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Systematic Analysis of Donor and Isolation Factor’s Impact on Human Islet Yield and Size Distribution

Y. Wang,1 K. K. Danielson,1 A. Ropski, T. Harvat, B. Barbaro, D. Paushter, M. Qi, and J. Oberholzer

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

Islet transplantation is a promising therapy for T1DM. Key factors influencing islet yield have been identified with conflicting results. In this study, we analyzed 276 isolations to identify variables for islet yield and, additionally, islet size and size distribution. Pearson correlation analyses demonstrated that BMI had a positive correlation with pancreas size, actual islet count (AIC), and islet equivalent (IEQ)/g (all p ≤ 0.009), while CIT had a negative correlation with AIC and IEQ/g (all p ≤ 0.003). In mixed linear regression, BMI also had a positive correlation with islet size but only for shorter digestion times (≤15 min); there was no association between BMI and islet size for longer digestion times (>15 min). CIT was not associated with islet size. Donor age, sex, and preservation solutions were shown to have no correlation with islet yields or size distribution. Pancreas size had a positive correlation with AIC and a negative association with IEQ/g; it also had positive association with islet size but only for females, not males. Overdigestion was positively associated with islet counts; however, there was also a greater proportion of smaller islets when digestion rate was >74% (p = 0.005). Of the three collagenases analyzed, Sigma V had the lowest digestion rate (mean = 65%), approximately 5% or 10% lower than Roche Liberase HI (p = 0.04) and Serva NB1 (p = 0.0003), respectively; however, the Sigma V group showed better islet size preservation. Yet, the enzymes resulted in similar IEQ/g digested tissue. Of the isolated islets, 70.2% were smaller than 150 μm and contributed only 20.4% to the total IEQ, while 7.4% of the islets were larger than 250 μm but contributed 42.4% to the total IEQ. In summary, BMI, pancreas size, and CIT are useful variables for predicting islet yield, but selection of enzyme and balancing digestion time and rate are also important.

Key words: Pancreatic islet of Langerhans; Islet isolation; Human islet transplantation; Actual islet count (AIC); Islet equivalent (IEQ)/g; Islet size; Size distribution

Received February 1, 2012; final acceptance October 10, 2012. Online prepub date: January 28, 2013.
1These authors provided equal contribution to this work.
Address correspondence to José Oberholzer, M.D., Department of Transplant/Surgery, University of Illinois at Chicago, 840 South Wood Street, Clinical Sciences Building, Suite 502, Chicago, IL 60612, USA. Tel: +1 312 996 6771; 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. 2335-2344, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X662381
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Technique of Endoscopic Biopsy of Islet Allografts Transplanted Into the Gastric Submucosal Space in Pigs

Minoru Fujita,*† Kevin M. McGrath,‡ Rita Bottino,*§ Eefje M. Dons,*¶ Cassandra Long,* Goutham Kumar,* Burcin Ekser,*# Gabriel J. Echeverri,** Jiro Hata,†† Ken Haruma,† David K. C. Cooper,* and Hidetaka Hara*

*Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA, USA
†Division of Gastroenterology, Department of Internal Medicine, Kawasaki Medical School, Kurashiki, Japan
‡Division of Gastroenterology, Hepatology and Nutrition, University of Pittsburgh, Pittsburgh, PA, USA
§Division of Immunogenetics, Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, USA
¶Department of Surgery, Erasmus Medical Center Rotterdam, Rotterdam, The Netherlands
#Department of Surgery, Transplantation and Advanced Technologies, Vascular Surgery and Organ Transplant Unit, University Hospital of Catania, Catania, Italy
**Transplantation Unit, Fundacion Clinica Valle del Lili, ICESI University School of Medicine, Cali, Colombia
††Division of Endoscopy and Ultrasonography, Department of Clinical Pathology and Laboratory Medicine, Kawasaki Medical School, Kurashiki, Japan

Currently, islet cells are transplanted into the liver via portal vein infusion. One disadvantage of this approach is that it is not possible to adequately biopsy the islets in the liver to assess for rejection. Islet transplantation (Tx) into the gastric submucosal space (GSMS) can be performed endoscopically and has the potential advantage of histological evaluation by endoscopic biopsy. The aim of this study was to determine whether a representative allograft sample could be obtained endoscopically. We performed islet Tx into the GSMS in nonimmunosuppressed pigs using simple endoscopic submucosal injection. Islets were transplanted at four sites. Endoscopic ultrasonography and biopsy of the transplanted islets at two sites by modified endoscopic submucosal dissection were carried out successfully in all pigs 5 days after islet Tx. Tissue obtained at both biopsy and necropsy (including full-thickness sections of the gastric wall around the sites of the remaining islets and biopsies) were examined by histology and immunohistochemistry to confirm the presence of the islet grafts and any features of rejection. Representative allograft sampling was successfully obtained from all biopsy sites. All biopsies included islets with insulin-positive staining. There was significant CD3+ and CD68+ cell infiltration in the islet masses obtained at biopsy and from sections taken at necropsy, with similar histopathological features. Endoscopic biopsy of islet allografts in the GSMS is feasible, provides accurate histopathological data, and would provide a significant advance if translated into clinical practice.

Key words: Endoscopy; Gastric submucosal space (GSMS); Biopsy of islets; Pig; Transplantation of islets

Received October 17, 2011; final acceptance October 18, 2012. Online prepub date: January 18, 2013.
Address correspondence to Hidetaka Hara, M.D., Ph.D., Research Assistant Professor, Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Starzl Biomedical Science Tower, W1558, 200 Lothrop Street, Pittsburgh, PA 15261, USA. Tel: +1-412-383-6960; Fax: +1-412-624-1172; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2345-2354, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657972
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Transplantation of Stem Cells Obtained From Murine Dental Pulp Improves Pancreatic Damage, Renal Function, and Painful Diabetic Neuropathy in Diabetic Type 1 Mouse Model

Elisalva Teixeira Guimarães,*† Gabriela da Silva Cruz,*‡ Tiago Farias de Almeida,* Bruno Solano de Freitas Souza,‡ Carla Martins Kaneto,‡ Juliana Fraga Vasconcelos,* Washington Luis Conrado dos Santos,* Ricardo Ribeiro-dos-Santos,*‡ Cristiane Flora Villarreal,*§ and Milena Botelho Pereira Soares*‡

*Centro de Pesquisas Gonçalo Moniz, FIOCRUZ, Bahia, Brazil
†Universidade do Estado da Bahia, Salvador, Bahia, Brazil
‡Centro de Biotecnologia e Terapia Celular, Hospital São Rafael, Salvador, Bahia, Brazil
§Universidade Federal da Bahia, Salvador, Bahia, Brazil

Diabetes mellitus (DM) is one of the most common and serious chronic diseases in the world. Here, we investigated the effects of mouse dental pulp stem cell (mDPSC) transplantation in a streptozotocin (STZ)-induced diabetes type 1 model. C57BL/6 mice were treated intraperitoneally with 80 mg/kg of STZ and transplanted with 1 × 106 mDPSCs or injected with saline, by an endovenous route, after diabetes onset. Blood and urine glucose levels were reduced in hyperglycemic mice treated with mDPSCs when compared to saline-treated controls. This correlated with an increase in pancreatic islets and insulin production 30 days after mDPSC therapy. Moreover, urea and proteinuria levels normalized after mDPSC transplantation in diabetic mice, indicating an improvement of renal function. This was confirmed by a histopathological analysis of kidney sections. We observed the loss of the epithelial brush border and proximal tubule dilatation only in saline-treated diabetic mice, which is indicative of acute renal lesion. STZ-induced thermal hyperalgesia was also reduced after cell therapy. Three days after transplantation, mDPSC-treated diabetic mice exhibited nociceptive thresholds similar to that of nondiabetic mice, an effect maintained throughout the 90-day evaluation period. Immunofluorescence analyses of the pancreas revealed the presence of GFP+ cells in, or surrounding, pancreatic islets. Our results demonstrate that mDPSCs may contribute to pancreatic β-cell renewal, prevent renal damage in diabetic animals, and produce a powerful and long-lasting antinociceptive effect on behavioral neuropathic pain. Our results suggest stem cell therapy as an option for the control of diabetes complications such as intractable diabetic neuropathic pain.

Key words: Stem cells; Dental pulp; Streptozotocin (STZ); Neuropathic pain; Hyperglycemia

Received January 19, 2011; final acceptance October 8, 2012. Online prepub date: October 12, 2012.
Address correspondence to Milena Botelho Pereira Soares, Centro de Pesquisas Gonçalo Moniz. Rua Waldemar Falcão, 121 - Candeal - Salvador, BA, Brazil, 40296-710. Tel: +55-71-3176-2292, ext. 260/272; Fax: +55-71-3176-2272; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2355-2366, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658737
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Low-Dose Rapamycin Unmasks the Protective Potential of Targeting Intragraft NF-kB for Islet Transplants

Nathan W. Zammit, Bernice M. Tan, Stacey N. Walters, David Liuwantara, Jeanette E. Villanueva, Elisabeth K. Malle, and Shane T. Grey

Gene Therapy and Autoimmunity Group, Immunology Program, Garvan Institute, Darlinghurst, NSW, Australia

Islet grafts can contribute to their own destruction via the elaboration of proinflammatory genes, many of which are transcriptionally regulated by nuclear factor κ-light-chain-enhancer of activated B-cells (NF-κB). Thus, NF-κB constitutes an enticing gene therapy candidate to improve the success of islet transplantation. To test this hypothesis in vivo, we blocked NF-κB in BALB/c (H2d) to C57/BL6 (H2b) mouse islet allografts by genetically engineering islets to express the NF-κB superrepressor, IκBα. Here we show by microarray and RTqPCR that islets exhibit an intrinsic early immediate proinflammatory response, with the most highly upregulated proinflammatory genes comprising the chemokines Cxcl1, Cxcl2, Cxcl10, and Ccl2; the cytokines Tnf-α and Il-6; and the adhesion molecule Icam1. Overexpression of IκBα inhibited the expression of these genes by 50–95% in islets and MIN6 β-cells in vitro, by inhibiting NF-κB-dependent gene transcription. Histological and RTqPCR analysis at postoperative day (POD) 10 revealed that IκBα-transduced islet allografts exhibited improved islet architecture and strong insulin-labeling with decreased Ccl2 and Il-6 mRNA levels compared to the GFP-transduced control grafts. Despite these protective effects, NF-κB-blocked islet allografts were promptly rejected in our MHC-mismatched mouse model. However, IκBα-expressing grafts did harbor localized “pockets” of Foxp3+ mononuclear cells not evident in the control grafts. This result suggested that the effect of the NF-κB blockade might synergize with regulatory T-cell-sparing rapamycin. Indeed, combining intragraft IκBα expression with low-dose rapamycin increased the mean survival time of islet allografts from 20 to 81 days, with 20% of the grafts surviving for greater than 100 days. In conclusion, rapamycin unmasks the protective potential of intragraft NF-κB blockade, which can, in some cases, permit permanent allograft survival without continuous systemic immunosuppression.

Key words: Inhibitor of κB protein (IκBα); Islet transplantation; Inflammation; Nuclear factor κ-light-chain-enhancer of activated B-cells (NF-κB); Rapamycin

Received July 10, 2012; final acceptance September 28, 2012. Online prepub date: October 31, 2012.
Address correspondence to Shane T. Grey, Gene Therapy and Autoimmunity Group, Immunology Program, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, NSW 2010, Australia. Tel: +011-61-2-9295-8104; Mobile: +0413-443-729; Fax: +011-61-2-9295-8404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2367-2380, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658881
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

A Critical Role for the TLR4/TRIF Pathway in Allogeneic Hematopoietic Cell Rejection by Innate Immune Cells

Hong Xu,*1 Jun Yan,†1 Ziqiang Zhu,* Lala-Rukh Hussain,* Yiming Huang,* Chuanlin Ding,† Larry D. Bozulic,* Yujie Wen,* and Suzanne T. Ildstad*

*Institute for Cellular Therapeutics, University of Louisville, Louisville, KY, USA
†James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA

We show for the first time that signaling through the TLR4/TRIF pathway plays a critical role in allogeneic bone marrow cell (BMC) rejection. This appears to be unique to BMCs as organ allografts are rejected mainly via MyD88 signaling. Using T- or T-/B-cell-deficient mice, we found that BMC allorejection occurred early before T-cell activation and was T- and B-cell independent, suggesting an effector role for innate immune cells in BMC rejection. We further demonstrated the innate immune signaling in BMC allorejection by showing superior engraftment in mice deficient in TRIF or TLR4 but not in MyD88 or TLR3. The restored cytotoxicity in TRIF-deficient recipients transferred with wild-type F4/80+ or NK1.1+ cells suggests TRIF signaling dependence on macrophages or NK cells in early BMC rejection. Production of the proinflammatory cytokine IL-6 and TRIF relevant chemokine MCP-1 was significantly increased early after bone marrow transplantation. In vivo specific depletion of macrophages or NK innate immune cells in combination with anti-CD154/rapamycin resulted in additive-enhanced allogeneic engraftment. The requirement for irradiation was completely eliminated when both macrophages and NK cells were depleted in combination with anti-CD154/rapamycin to target T- and B-cells, supporting the hypothesis that two barriers involving innate and adaptive immunity exist in mediating the rejection of allogeneic BMCs. In summary, our results clearly demonstrate a previously unappreciated role for innate immunity in BMC allorejection via signaling through a unique MyD88-independent TLR4/TRIF mechanism. These findings may have direct clinical impact on strategies for conditioning recipients for stem cell transplantation.

Key words: TLR4; TRIF; Bone marrow transplantation; Innate immunity

Received May 31, 2012; final acceptance October 11, 2012. Online prepub date: November 8, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Suzanne T. Ildstad, M.D., Director, Institute for Cellular Therapeutics, Jewish Hospital Distinguished Professor of Transplantation, Professor of Surgery, University of Louisville, 570 South Preston Street, Suite 404, Louisville, KY 40202-1760, USA. Tel: +1-502-852-2080; Fax: 1-502-852-2079; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2381-2392, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658863
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Angiogenesis Following Cell Injection Is Induced by an Excess Inflammatory Response Coordinated by Bone Marrow Cells

Hidemi Hattori,* Yoshiko Amano,* Yoshiko Habu-Ogawa,† Takahiro Ando,‡ Bonpei Takase,§ and Masayuki Ishihara*

*Division of Biomedical Engineering, Research Institute, National Defense Medical College, Saitama, Japan
†Department of Immunology and Microbiology, National Defense Medical College, Saitama, Japan
‡Department of Electronics and Electrical Engineering, Keio University, Yokohama, Kanagawa, Japan
§Department of Intensive Care Medicine, National Defense Medical College, Saitama, Japan

The aim of this study was to identify novel angiogenic mechanisms underlying the regenerative process. To that end, interactions between adipose tissue-derived stromal cells (ASCs) and bone marrow cells (BMCs) were initially investigated using real-time fluorescence optical imaging. To monitor cell behavior in mice, we injected green fluorescent protein-positive (GFP+) BMCs into the tail vein and injected PKH26-labeled ASCs behind the ears. Angiogenesis and inflammation were observed at these sites via an optical imaging probe. Injected GFP+ BMCs migrated from the blood vessels into the tissues surrounding the ASC injection sites. Many of the migrating GFP+ BMCs discovered at the ASC injection sites were inflammatory cells, including Gr-1+, CD11b+, and F4/80+ cells. ASCs cocultured with inflammatory cells secreted increased levels of chemokines such as macrophage inflammatory protein (MIP)-1α, MIP-1β, keratinocyte-derived chemokines, and monocyte chemotactic protein 1. Similarly, these ASCs secreted increased levels of angiogenic growth factors such as hepatocyte growth factor and vascular endothelial growth factor. However, when anti-CXC chemokine receptor type 4 antibody was injected at regular intervals, the migration of GFP+ BMCs (especially Gr-1+ and CD11b+ cells) to ASC injection sites was inhibited, as was angiogenesis. The collective influence of the injected ASCs and BMC-derived inflammatory cells promoted acute inflammation and angiogenesis. Together, the results suggest that the outcome of cell-based angiogenic therapy is influenced not only by the injected cells but also by the effect of intrinsic inflammatory cells.

Key words: Adipose tissue-derived stromal cells; Angiogenesis; Imaging; Inflammation; CXCR4

Received January 9, 2012; final acceptance October 19, 2012. Online prepub date: November 8, 2012.
Address correspondence to Hidemi Hattori, Ph.D., Division of Biomedical Engineering, Research Institute, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan. Tel: +81-42-995-1618; Fax: +81-42-991-1611; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2393-2408, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658854
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

BMP2 Is Superior to BMP4 for Promoting Human Muscle-Derived Stem Cell-Mediated Bone Regeneration in a Critical-Sized Calvarial Defect Model

Xueqin Gao,*1 Arvydas Usas,*1 Aiping Lu,* Ying Tang,*† Bing Wang,*† Chien-Wen Chen,* Hongshuai Li,* Jessica C. Tebbets,* James H. Cummins,* and Johnny Huard*

*Stem Cell Research Center, Growth and Developmental Laboratory, Department of Orthopaedic Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA
†Molecular Therapy Laboratory, Department of Orthopaedic Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

Muscle-derived cells have been successfully isolated using a variety of different methods and have been shown to possess multilineage differentiation capacities, including an ability to differentiate into articular cartilage and bone in vivo; however, the characterization of human muscle-derived stem cells (hMDSCs) and their bone regenerative capacities have not been fully investigated. Genetic modification of these cells may enhance their osteogenic capacity, which could potentially be applied to bone regenerative therapies. We found that hMDSCs, isolated by the preplate technique, consistently expressed the myogenic marker CD56, the pericyte/endothelial cell marker CD146, and the mesenchymal stem cell markers CD73, CD90, CD105, and CD44 but did not express the hematopoietic stem cell marker CD45, and they could undergo osteogenic, chondrogenic, adipogenic, and myogenic differentiation in vitro. In order to investigate the osteoinductive potential of hMDSCs, we constructed a retroviral vector expressing BMP4 and GFP and a lentiviral vector expressing BMP2. The BMP4-expressing hMDSCs were able to undergo osteogenic differentiation in vitro and exhibited enhanced mineralization compared to nontransduced cells; however, when transplanted into a calvarial defect, they failed to regenerate bone. Local administration of BMP4 protein and cell pretreatment with N-acetylcysteine (NAC), which improves cell survival, did not enhance the osteogenic capacity of the retro-BMP4-transduced cells. In contrast, lenti-BMP2-transduced hMDSCs not only exhibited enhanced in vitro osteogenic differentiation but also induced robust bone formation and nearly completely healed a critical-sized calvarial defect in CD-1 nude mice 6 weeks following transplantation. Herovici’s staining of the regenerated bone demonstrated that the bone matrix contained a large amount of type I collagen. Our findings indicated that the hMDSCs are likely mesenchymal stem cells of muscle origin and that BMP2 is more efficient than BMP4 in promoting the bone

regenerative capacity of the hMDSCs in vivo.

Key words: Human muscle-derived stem cells (hMDSCs); Lentiviral vector; Bone morphogenetic protein 2 (BMP2); Bone morphogenetic protein 4 (BMP4); Retroviral vector; Calvarial defect

Received March 16, 2012; final acceptance October 18, 2012. Online prepub date: November 1, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Johnny Huard, Stem Cell Research Center, Growth and Developmental Laboratory, Department of Orthopaedic Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15219, USA. Tel: +1-412-648-2789; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 2409-2420, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658746
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Effects of Exercise Interventions in Graft-Versus-Host Disease Models

Carmen Fiuza-Luces,* África González-Murillo,† Luisa Soares-Miranda,‡ Jesús Martínez Palacio,§ Isabel Colmenero,† Fernando Casco,† Gustavo Melén,† María Morán,¶#  Alejandro Lucia,*1 and Manuel Ramírez†1

*Universidad Europea and Instituto de Investigación (i+12), Madrid, Spain
†Hospital Infantil Universitario Niño Jesús, Madrid, Spain
‡Research Centre in Physical Activity, Health and Leisure, Faculty of Sport, University of Porto, Porto, Portugal
§Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT), Madrid, Spain
¶Laboratorio de Enfermedades Raras: Mitocondriales y Neuromusculares, Instituto de Investigación Hospital Universitario, Madrid, Spain
#Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), U723, Valencia, Spain

Graft-versus-host-disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT), which is associated with high morbimortality and decreased patients’ physical capacity. We evaluated the effects of an 11-week moderate-intensity exercise (treadmill) training program performed after allo-HSCT in a murine acute (aGVHD) and chronic GVHD model (cGVHD). Female mice (aged 8–12 weeks) were randomly assigned to the exercise or the control group. They completed a maximal treadmill test before allo-HSCT (with donor bone marrow cells and splenocytes) and after the 11-week period, during which we evaluated clinical severity scores and survival (Kaplan–Meier method). Before allo-HSCT and at days +21, +52 and +83 (upon sacrifice), we collected blood samples for immune cell reconstitution and cytokine analysis. The main results were that (i) in aGVHD, exercise improved maximal physical capacity over the 11-week period compared with pre-allo-HSCT conditions (p < 0.001 for the between-group comparison) and benefited total clinical score evolution (p = 0.05 for the group × time interaction effect), without altering immune reconstitution; (ii) in cGVHD, exercise training resulted in a lesser deterioration of physical capacity after 11 weeks (p = 0.023). Our results highlight the potential beneficial effects of exercise as coadjuvant intervention against GVHD, especially in the acute form of the disease.

Key words: Physical activity; Animal models; Inflammation; Immunity; Survival

Received June 13, 2012; final acceptance August 30, 2012. Online prepub date: October 31, 2012.
1Alejandro Lucia and Manuel Ramírez share senior authorship.
Address correspondence to Alejandro Lucia, Universidad Europea de Madrid, 28670 Villaviciosa de Odón, Madrid, Spain. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it