Cell Transplantation 21(5) Abstracts

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Cell Transplantation, Vol. 21, pp. 801–814, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X 627507
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Current Applications of Human Pluripotent Stem Cells: Possibilities and Challenges

Pai-Jiun Ho,* Men-Luh Yen,† Shaw-Fang Yet,‡ and B. Linju Yen*

*Regenerative Medicine Research Group, Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan
†Departmant of Primary Medicine and Department of Obstetrics/Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan
‡Cardiovascular Research Group, Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan

Stem cells are self-renewable cells with the differentiation capacity to develop into somatic cells with biological functions. This ability to sustain a renewable source of multi- and/or pluripotential differentiation has brought new hope to the field of regenerative medicine in terms of cell therapy and tissue engineering. Moreover, stem cells are invaluable tools as in vitro models for studying diverse fields, from basic scientific questions such as developmental processes and lineage commitment, to practical application including drug screening and testing. The stem cells with widest differentiation potential are pluripotent stem cells (PSCs), which are rare cells with the ability to generate somatic cells from all three germ layers. PSCs are considered the most optimal choice for therapeutic potential of stem cells, bringing new impetus to the field of regenerative medicine. In this article, we discuss the therapeutic potential of human PSCs (hPSCs) including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), reviewing the current preclinical and clinical data using these stem cells. We describe the classification of different sources of hPSCs, ongoing research, and currently encountered clinical obstacles of these novel and versatile human stem cells.

Key words: Pluripotent stem cells (PSCs); Human embryonic stem cells (hESCs); Induced pluripotent stem cells (iPSCs); Differentiation; Cell therapy; Clinical trial; Drug development

Received March 10, 2011; final acceptance May 28, 2011. Online prepub date: March 22, 2012.
Address correspondence to B. Linju Yen, Regenerative Medicine Research Group, Institute of Cellular and System Medicine, National Health Research Institutes, No. 35 Keyan Road , Zhunan, 350, Taiwan. Tel: +886-37-246-166, x37518; Fax: +886-37-587-408; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 815–825, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X601019
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Generation of Feeder-Free Pig Induced Pluripotent Stem Cells Without Pou5f1

Nuria Montserrat,* Lorena de Oñate,* Elena Garreta,* Federico González,* Antonio Adamo,* Cristina Eguizábal,* Sophia Häfner,* Rita Vassena,* and Juan Carlos Izpisua Belmonte*†

*Center for Regenerative Medicine in Barcelona, Dr. Aiguader, 88, 08003, Barcelona, Spain
†Gene Expression Laboratory, The Salk Institute for Biological Studies, La Jolla, CA, USA

The pig represents an ideal large-animal model, intermediate between rodents and humans, for the preclinical assessment of emerging cell therapies. As no validated pig embryonic stem (pES) cell lines have been derived so far, pig induced pluripotent stem cells (piPSCs) should offer an alternative source of undifferentiated cells to advance regenerative medicine research from bench to clinical trial. We report here for the first time the derivation of piPSCs from adult fibroblast with only three transcription factors: Sox2 (sex determining region Y-box 2), Klf4 (Krüppel-like factor 4), and c-Myc (avian myelocytomatosis viral oncogene homolog). We have been able to demonstrate that exogenous Pou5f1 (POU domain class 5 transcription factor 1; abbreviated as Octamer-4: Oct4) is dispensable to achieve and maintain pluripotency in the generation of piPSCs. To the best of our knowledge, this is also the first report of somatic reprogramming in any species without the overexpression, either directly or indirectly, of Oct4. Moreover, we were able to generate piPSCs without the use of feeder cells, approaching thus xeno-free conditions. Our work paves the way for the derivation of clinical grade piPSCs for regenerative medicine.

Key words: Cell culture; Induced pluripotent stem (iPS) cells; Embryonic stem cells (ESCs); Clinical translation

Address correspondence to Juan Carlos Izpisua Belmonte, Center for Regenerative Medicine in Barcelona, Dr. Aiguader, 88, 08003, Barcelona, Spain. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Gene Expression Laboratory, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 827–843, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X593163
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Cotransplantation of Human Embryonic Stem Cell-Derived Neural Progenitors and Schwann Cells in a Rat Spinal Cord Contusion Injury Model Elicits a Distinct Neurogenesis and Functional Recovery

Ali Niapour,*†‡ Fereshteh Karamali,* Shiva Nemati,§ Zahra Taghipour,*† Mohammad Mardani,† Mohammad Hossein Nasr-Esfahani,* and Hossein Baharvand§¶

*Department of Cell and Molecular Biology, Cell Science Research Center, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran
†Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Science, Isfahan, Iran
‡Department of Anatomical Sciences, Ardebil University of Medical Science, Ardebil, Iran
§Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
¶Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran

Cotransplantation of neural progenitors (NPs) with Schwann cells (SCs) might be a way to overcome low rate of neuronal differentiation of NPs following transplantation in spinal cord injury (SCI) and the improvement of locomotor recovery. In this study, we initially generated NPs from human embryonic stem cells (hESCs) and investigated their potential for neuronal differentiation and functional recovery when cocultured with SCs in vitro and cotransplanted in a rat acute model of contused SCI. Cocultivation results revealed that the presence of SCs provided a consistent status for hESC-NPs and recharged their neural differentiation toward a predominantly neuronal fate. Following transplantation, a significant functional recovery was observed in all engrafted groups (NPs, SCs, NPs + SCs) relative to the vehicle and control groups. We also observed that animals receiving cotransplants established a better state as assessed with the BBB functional test. Immunohistofluorescence evaluation 5 weeks after transplantation showed invigorated neuronal differentiation and limited proliferation in the cotransplanted group when compared to the individual hESC-NPgrafted group. These findings have demonstrated that the cotransplantation of SCs with hESC-NPs could offer a synergistic effect, promoting neuronal differentiation and functional recovery.

Key words: Rat Schwann cell; Human neural progenitor; Coculture; Cotransplantation; Differentiation; Spinal cord injury

Address correspondence to Hossein Baharvand, Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 19395-4644, Tehran, Iran. Tel: +98 21 22306485; Fax: +98 21 22310406; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Mohammad Hossein, Nasr-Esfahani Department of Cell and Molecular Biology, Cell Science Research Center, Royan Institute for Animal Biotechnology, P.O. Box 815896-8433, Isfahan, Iran. Tel: +98 311 26129003; Fax: +98 311 2602555l; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 845–856, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627417
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Therapeutic Benefit of Treatment of Stroke With Simvastatin and Human Umbilical Cord Blood Cells: Neurogenesis, Synaptic Plasticity, and Axon Growth

Xu Cui,* Michael Chopp,*† Amjad Shehadah,* Alex Zacharek,* Nicole Kuzmin-Nichols,‡ Cyndy Davis Sanberg,‡ Junhao Dai,* Chunling Zhang,* Yuji Ueno,* Cynthia Roberts,* and Jieli Chen*

*Department of Neurology, Henry Ford Hospital, Detroit, MI, USA
†Department of Physics, Oakland University, Rochester, MI, USA
‡Saneron CCEL Therapeutics, Inc., Tampa, FL, USA

The therapeutic efficacy of cell-based therapy after stroke can be enhanced by making the host brain tissue more receptive to the administered cells, which thereby facilitates brain plasticity. We hypothesized that simvastatin increases human umbilical cord blood cell (HUCBC) migration into the ischemic brain and promotes brain plasticity and neurological functional outcome after stroke. Rats were subjected to 2-h middle cerebral artery occlusion (MCAo) and administered subtherapeutic doses of simvastatin (0.5 mg/kg, gavaged daily for 7 days), HUCBCs (1 × 106, one time injection via tail vein), or combination simvastatin with HUCBCs starting at 24 h after stroke. Combination treatment of stroke showed an interactive effect in improvement of neurological outcome compared with simvastatin or HUCBC monotherapy groups. In addition, combination treatment significantly increased brain-derived neurotrophic factor/TrkB expression and the number of engrafted HUCBCs in the ischemic brain compared with HUCBC monotherapy. The number of engrafted HUCBCs was significantly correlated with functional outcome (modified neurological severity score). Combination treatment significantly increased neurogenesis and synaptic plasticity in the ischemic brain, and promoted neuroblast migration in cultured subventricular zone explants. Using primary cultured neurons (PCNs), we found that combination treatment enhanced neurite outgrowth compared with nontreatment control, simvastatin or HUCBC supernatant monotherapy. Inhibition of TrkB significantly attenuated combination treatment-induced neurite outgrowth. Our data indicate that combination simvastatin and HUCBC treatment of stroke increases BDNF/TrkB expression, enhances HUCBC migration into the ischemic brain, amplifies endogenous neurogenesis, synaptic plasticity and axonal growth, and thereby improves functional outcome after stroke.

Key words: Simvastatin; Human umbilical cord blood cells (HUCBCs); Neurogenesis; Synaptic plasticity; Stroke

Address correspondence to Jieli Chen, Department of Neurology, E&R Bldg., Room 3091, Henry Ford Hospital, 2799 W Grand Blvd., Detroit, MI 48202, USA. Tel: 313-916-1991; Fax: 313-916-1318; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 857–869, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X612486
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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An Effective and Safe Supplement for Stem Cells Expansion Ex Vivo: Cord Blood Serum

Hai-ying Ma,* Li Yao,† Yan-qiu Yu,* Li Li,* Ling Ma,* Wen-juan Wei,* Xiao-mei Lu,* Li-li Du,* and Yu-nan Jin*

*Department of Pathophysiology, College of Basic Medical Sciences, China Medical University, Shenyang, China
†Department of Nephrology, The First Hospital of China Medical University, Shenyang, China

Mesenchymal stem cells (MSCs) are potential and optimal stem cells in clinical cell therapy, and fetal bovine serum (FBS) is widely used for expansion of MSCs, despite the risks of infectious disease transmission and immunological reaction of the xenogenic origin. This study was designed to compare human four blood group cord blood serum (CBS) with FBS in culturing human placenta-derived mesenchymal stem cells (hPDMSCs), which were derived from four blood group donors. The expansion medium included: 10% FBS, 10% A-CBS, 10% B-CBS, 10% O-CBS, and 10% AB-CBS. Cumulative population doubling, generation time, fold expansion rates and differentiation capacity, cell cycle, and immunophenotype were also assessed. The results showed that fold expansion rate and cumulative population doubling of hPDMSCs significantly increased during long-term MSC expansion in CBS medium, but the generation time decreased compared with FBS. CBS might be an effective supplement for stem cells expand rapidly ex vivo. Cell cycle and differentiation assays showed that most of the hPDMSCs expanding in the presence of CBS were in stationary phase, which was the characteristic of stem cells, and they retained their ability to differentiate into chondrogenic and endothelial cells. By comparing these four blood groups of CBS, we found that there was no significant difference among different blood groups in culturing hPDMSCs, which were isolated from different blood group donors. So CBS may be an optimal replacement to avoid the risks of FBS application in expansion of stem cell for clinical cell therapy and tissue engineering.

Key words: Cord blood serum (CBS); Fetal bovine serum (FBS); Mesenchymal stem cells (MSCs); Cell expansion; Placenta

Address correspondence to Yanqiu Yu, Department of Pathophysiology, College of Basic Medical Sciences, China Medical University, Shenyang, 110001, China. Tel/fax: 86 24 23256666, ext. 5362; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 871–887, 2012
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DOI: http://dx.doi.org/10.3727/096368911X623871
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Manipulation of the Recipient Retinal Environment by Ectopic Expression of Neurotrophic Growth Factors Can Improve Transplanted Photoreceptor Integration and Survival

E. L. West,* R. A. Pearson,* Y. Duran,* A. Gonzalez-Cordero,* R. E. MacLaren,*†1 A. J. Smith,* J. C. Sowden,‡ and R. R. Ali*§

*Department of Genetics, University College London Institute of Ophthalmology, London, UK
†Vitreoretinal Service, Moorfields Eye Hospital, London, UK
‡Developmental Biology Unit, University College London Institute of Child Health, London, UK
§Molecular Immunology Unit, University College London Institute of Child Health, London, UK

Degeneration of the neural retina is the leading cause of untreatable blindness in the developed world. Stem cell replacement therapy offers a novel strategy for retinal repair. Postmitotic photoreceptor precursors derived from the early postnatal (P) retina are able to migrate and integrate into the adult mouse retina following transplantation into the subretinal space, but it is likely that a large number of these cells would be required to restore vision. The adult recipient retina presents a very different environment to that from which photoreceptor precursor donor cells isolated from the developing postnatal retina are derived. Here we considered the possibility that modulation of the recipient environment by ectopic expression of developmentally regulated growth factors, normally present during photoreceptor development, might enhance the migration and integration of transplanted cells into the adult neural retina. Adeno-associated viral (AAV) vectors were used to introduce three growth factors previously reported to play a role in photoreceptor development, IGF1, FGF2, and CNTF, into the adult retina, prior to transplantation of P4 cells derived from the Nrl.GFP+ve neural retina. At 3 weeks posttransplantation the number of integrated, differentiated photoreceptor cells present in AAV-mediated neurotrophic factor-treated eyes was assessed and compared to control treated contralateral eyes. We show, firstly, that it is possible to manipulate the recipient retinal microenvironment via rAAV-mediated gene transfer with respect to these developmentally relevant growth factors. Moreover, when combined with cell transplantation, AAV-mediated expression of IGF1 led to significantly increased levels of cell integration, while overexpression of FGF2 had no significant effect on integrated cell number. Conversely, expression of CNTF led to a significant decrease in cell integration and an exacerbated glial response that led to glial scarring. Together, these findings demonstrate the importance of the extrinsic environment of the recipient retina for photoreceptor cell transplantation and show for the first time that it is possible to manipulate this environment using viral vectors to influence photoreceptor transplantation efficiency.

Key words: Photoreceptor; Retina; Transplantation; Neurotrophic factors; Gene therapy; Stem cell

1Current address: Nuffield Laboratory of Ophthalmology, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, UK.
Address correspondence to Rachael A. Pearson, Department of Genetics, University College London Institute of Ophthalmology, 11-43 Bath Street, London, EC1V 9EL, UK. Tel: +44 (0)20 7608 6902; Fax: +44 (0)20 7608 6903; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Robin R. Ali, Department of Genetics, University College London Institute of Ophthalmology, 11-43 Bath Street, London, EC1V 9EL, UK. Tel: +44 (0)20 7608 6902; Fax: +44 (0)20 7608 6903; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 889–900, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X603639
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Evidence of Endoplasmic Reticulum Stress Mediating Cell Death in Transplanted Human Islets

Sarita Negi,*† Soon Hyang Park,*†1 Arif Jetha,*† Reid Aikin,*† Michel Tremblay,‡ and Steven Paraskevas*†

*Human Islet Transplantation Laboratory, McGill University Health Centre, Montreal, Quebec, Canada
†Departments of Surgery, McGill University, Montreal, Quebec, Canada
‡Goodman Cancer Center, McGill University, Montreal, Quebec, Canada

A key limitation to the success of islet transplantation is islet cell exhaustion and cell death during islet isolation and following transplantation. Endoplasmic reticulum (ER) stress has been identified as an important mechanism in the development of β-cell dysfunction, cell death, and diabetes. This study investigated the role of ER stress in islet loss during human islet isolation and posttransplantation in a diabetic athymic mouse model. Islets were isolated from human organ donor pancreata using intraductal enzymatic dissociation and continuous density gradient purification. ER stress mediators were assessed by Western blot and by RT-PCR. Caspase-3 activity was quantified by a bioluminescent peptide cleavage assay. Normal and streptozotocin-treated diabetic nude mice were transplanted with 2,000 IEQ of human islets under the kidney capsule and the grafts were harvested 3 or 28 days after transplantation. The grafts were analyzed for the presence for ER stress signals by immunohistochemistry. Isolated islets demonstrated higher levels of ER chaperone Bip, ER stress mediators eIF2α, ATF, spliced XBP-1, and CHOP, and also ER stress-associated apoptotic signals like JNK, caspase-3/7, and cleaved PARP. Donor pancreatic tissue did not show expression of any of these ER stress mediators. After transplantation, low expression of only protective ER stress mediators was evident in the grafts from the normal recipients. In contrast, both protective and apoptotic ER stress mediators were highly expressed in the grafts of hyperglycemic mice. ER stress mediators were induced during islet isolation and may contribute to islet apoptosis and cell death. Islet isolation activates ER stress and apoptotic pathways in isolated islets. Hyperglycemia may prolong this ER stress signal in engrafted islets, converting the protective aspects of the ER stress response to a proapoptotic response and thus contribute to deterioration of β-cell function and survival.

Key words: Islet; Human; Endoplasmic reticulum (ER) stress; Apoptosis; Transplantation

1Current affiliation: Biomedical Engineering, McGill University, Montreal, Quebec, Canada.
Address correspondence to Steven Paraskevas, M.D., Ph.D., S10.30, 687 Pine Avenue, W., McGill University Health Centre, Montreal, QC, H3A 1A1 Canada. Tel: (514) 934-1934, ext. 36522; Fax: (514) 843 1503; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 901–908, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X612468
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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HLA Class I Sensitization in Islet Transplant Recipients: Report From the Collaborative Islet Transplant Registry1

Bashoo Naziruddin,* Steve Wease,† Donald Stablein,† Franca B. Barton,† Thierry Berney,‡ Michael R. Rickels,§ and Rodolfo Alejandro¶

*Baylor Simmons Transplant Institute, Dallas, TX, USA
†The EMMES Corporation, Rockville, MD, USA
‡Service of Transplantation and Visceral Surgery, Geneva University Hospital and Medical School, Geneva, Switzerland
§Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
¶The Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL, USA

Pancreatic islet transplantation is a promising treatment option for patients severely affected with type 1 diabetes. This report from CITR presents pre- and posttransplant human leukocyte antigen (HLA) class I sensitization rates in islet-alone transplantation. Data came from 303 recipients transplanted with islet-alone between January 1999 and December 2008. HLA class I sensitization was determined by the presence of anti-HLA class I antibodies. Panel-reactive antibodies (PRA) from prior to islet infusion and at 6 months, and yearly posttransplant was correlated to measures of islet graft failure. The cumulative number of mismatched HLA alleles increased with each additional islet infusion from a median of 3 for one infusion to 9 for three infusions. Pretransplant PRA was not predictive of islet graft failure. However, development of PRA >20% posttransplant was associated with 3.6-fold (p < 0.001) increased hazard ratio for graft failure. Patients with complete graft loss who had discontinued immunosuppression had significantly higher rate of PRA ≥ 20% compared to those with functioning grafts who remained on immunosuppression. Exposure to repeat HLA class I mismatch at second or third islet infusions resulted in less frequent development of de novo HLA class I antibodies when compared to increased class I mismatch. The development of HLA class I antibodies while on immunosuppression is associated with subsequent islet graft failure. The risk of sensitization may be reduced by minimizing the number of islet donors used per recipient, and in the absence of donor-specific anti-HLA antibodies, repeating HLA class I mismatches with subsequent islet infusions.

Key words: Islet transplantation; HLA sensitization; HLA class I; Panel reactive antibodies

1The list of Collaborative Islet Transplant Registry (CITR) contributors is provided in the Acknowledgments.
Address correspondence to Bashoo Naziruddin, Ph.D., Islet Cell Laboratory, Transplant Services, Baylor University Medical Center, 3410 Worth Street, Dallas, TX 75246, USA. Tel: (214) 820-2662; Fax: (214) 820-7142; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 909–918, 2012
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DOI: http://dx.doi.org/10.3727/096368911X623934
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Attenuation of Murine Graft-Versus-Host Disease by a Tea Polyphenol

Jun Kanamune,* Yasuhiro Iwanaga,* Tatsuo Kina,† Hirofumi Noguchi,‡ Kazuaki Matsumura,§ Shinji Uemoto,¶ and Suong-Hyu Hyon#

*Transplant Unit, Kyoto University Hospital, Kyoto, Japan
†Department of Immunology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
‡Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, TX, USA
§Department of Material Science, JAIST, Ishikawa Prefecture, Japan
¶Division of Hepato-Biliary-Pancreatic and Transplant Surgery, Department of Surgery, Kyoto University, Kyoto, Japan
#Department of Simulation Medical Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan

Since donor T-cells’ allorecognition of host antigens is a prerequisite for the onset of graft-versus-host disease (GVHD), blocking their cellular signaling pathways can decrease the severity of GVHD. We hypothesized that epigallocatechin-3-gallate (EGCG), due to its strong affinity to macromolecules, would adhere to surface molecules of donor T cells, inhibit their allorecognition, and attenuate GVHD in the recipient. We tested the hypothesis by treating donor splenocytes with EGCG in both in vitro and in vivo murine GVHD models. EGCG treatment decreased the proliferation of donor cells in MLR cultures and secretion of IL-2 and INF-γ. It also reduced the epitope detection of CD3ε, CD4, and CD28 but did not downregulate the protein expression of these molecules, suggesting blockage of cell surface stimulatory signals. Similarly, EGCG treatment did not decrease mRNA expression for some of these molecules but decreased mitogeninduced cell proliferation, indicating that EGCG did not interfere the transcription of these genes but affected cell proliferation pathways. Furthermore, EGCG-treated donor splenocytes, when transplanted into immunocompromized recipient mice, decreased of proliferation, and the treatment extended the recipients’ survival at least during the early stage of GVHD. These results strongly suggest that EGCG attenuates GVHD by both blocking specific cell surface molecules and affecting the donor T-cell proliferation pathways.

Key words: Graft-versus-host disease (GVHD); Epigallocatechin-3-gallate (EGCG); T cells; Tea polyphenol

Address correspondence to Dr. Jun Kanamune, Transplant Unit, Kyoto University Hospital, 54 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan. Tel: +81-75-751-4322; Fax: +81-75-751-3665; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 919–931, 2012
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DOI: http://dx.doi.org/10.3727/096368911X623817
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Transplanted Fibroblasts Prevents Dysfunctional Repair in a Murine CXCR3-Deficient Scarring Model

Cecelia C. Yates, Diana Whaley, and Alan Wells

Department of Pathology and McGowan Institute for Regenerative Medicine, University of Pittsburgh and Pittsburgh VAMC, Pittsburgh, PA, USA

In skin, the regeneration of the ontogenically distinct mesenchymal and epithelial compartments must proceed in a coordinated manner orchestrated by extracellular signaling networks. We have recently found that the switch from regeneration to remodeling during repair is modulated by chemokines that bind CXCR3 receptor. If this signaling is disrupted wounds continue to be active, resulting in a chronic hypercellular and hypertrophic state characterized by an immature matrix composition. As healing is masterminded in large part by fibroblasts and their synthesis of the extracellular matrix, the question arose as to whether this ongoing scarring can be modulated by transplanted fibroblasts. We examined wounds in the CXCR3−/− mouse scarring model. These wounds exhibited a significant delay in healing in all areas compared to young and aged wild-type mice. Full-thickness wounds were transplanted with fibroblasts derived from newborn CXCR3−/− or wild-type mice. The transplanted fibroblasts were labeled with fluorescent dye (CM-DiI) and suspended in hyaluronic acid gel; by 30 days, these transplanted cells comprised some 30% of the dermal stromal cells regardless of the host or source of transplanted cells. Wild-type fibroblasts transplanted into CXCR3−/− mice wounds reversed the delay and dysfunction previously seen in CXCR3−/− wounds; this correction was not noted with transplanted CXCR3−/− fibroblasts. Additionally, transplant of CXCR3−/− cells into wounds in wild-type animals did not adversely affect those wounds. The transplanted fibroblasts exhibited strong survival and migration patterns and led to an increase in tensile strength. Expression of matrix proteins and collagen in CXCR3−/− wounds transplanted with wild-type fibroblasts resembled normal wild-type healing, and the wound matrix in wild-type mice transplanted with CXCR3−/− cells also presented a mature matrix. These suggest that the major determinant of healing versus scarring lies with the nature of the matrix. These findings have intriguing implications for rational cellular interventions aimed at promoting wound healing via cell therapy.

Key words: Extracellular matrix; Cell therapy; Collagen; Fibroblast; Epidermal–dermal communication

Address correspondence to Alan Wells, M.D., D.MSc., Department of Pathology, University of Pittsburgh, 3550 Terrace St., Scaife Hall, S-713, Pittsburgh, PA 15261, USA. Tel: 412-647-7813; Fax: 412-647-8567; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 933–942, 2012
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DOI: http://dx.doi.org/10.3727/096368911X600993
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Stem Cells in Burn Eschar

Vincent C. van der Veen,* Marcel Vlig,* Florine J. van Milligen,† Sharon I. de Vries,* Esther Middelkoop,*‡ and Magda M. W. Ulrich*‡

*Association of Dutch Burn Centers, Beverwijk, The Netherlands
†Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands
‡Department of Plastic, Reconstructive and Hand Surgery, VU University Medical Center, Amsterdam, The Netherlands

This study compares mesenchymal cells isolated from excised burn wound eschar with adipose-derived stem cells (ASCs) and dermal fibroblasts in their ability to conform to the requirements for multipotent mesenchymal stem cells (MSCs). A population of multipotent stem cells in burn eschar could be an interesting resource for tissue engineering approaches to heal burn wounds. Cells from burn eschar, dermis, and adipose tissue were assessed for relevant CD marker profiles using flow cytometry and for their trilineage differentiation ability in adipogenic, osteogenic, and chondrogenic conditions. Although the different cell types did not differ significantly in their CD marker expression, the eschar-derived cells and ASCs readily differentiated into adipocytes, osteoblasts, and chondrocytes, while dermal fibroblasts only exhibited some chondrogenic potential. We conclude that the eschar-derived mesenchymal cells represent a population of multipotent stem cells. The origin of the cells from burn eschar remains unclear, but it is likely they represent a population of adult stem cells mobilized from other parts of the body in response to the burn injury. Their resemblance to ASCs could also be cause for speculation that in deep burns the subcutaneous adipose tissue might be an important stem cell source for the healing wound.

Key words: Burns; Wound healing; Scarring; Mesenchymal stem cells (MSCs); Adipose-derived stem cells (ASCs); α-Smooth muscle actin

Address correspondence to Madga M. W. Ulrich, Association of Dutch Burn Centres, Postbus 1015, 1940 EA Beverwijk, The Netherlands. Tel: +31 251275506; Fax: 0031 251264948; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 943–958, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627453
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Allogenous Tendon Stem/Progenitor Cells in Silk Scaffold for Functional Shoulder Repair

Weiliang Shen,*†1 Jialin Chen,*‡1 Zi Yin,*‡ Xiao Chen,*‡ Huanhuan Liu,*‡ Boon Chin Heng,§ Weishan Chen,† and Hong-Wei Ouyang*‡

*Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, Hangzhou, China
†Department of Orthopedic Surgery, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China
‡Department of Sports Medicine, School of Medicine, Zhejiang University, Hangzhou, China
§School of Materials Science & Engineering, Nanyang Technological University, Singapore

Tendon stem/progenitor cells (TSPCs) were recently identified within tendon tissues. The aim of this study was to investigate TSPC-seeded knitted silk–collagen sponge scaffold for functional shoulder repair. The multidifferentiation potential, proliferation, and immune properties of TSPCs were investigated in vitro, while the efficacy of TSPC-seeded knitted silk–collagen sponge scaffolds in promoting rotator cuff regeneration was evaluated in vivo within a rabbit model. TSPCs, which exhibited universal stem cell characteristics (i.e., clonogenicity, high proliferative capacity, and multidifferentiation potential), nonimmunogenicity, and immunosuppression, proliferated well on our scaffold in vitro. Implantation of allogenous TSPC-seeded scaffolds within a rabbit rotator cuff injury model did not elicit an immunological reaction, but instead increased fibroblastic cell ingrowth and reduced infiltration of lymphocytes within the implantation sites at 4 and 8 weeks postsurgery. After 12 weeks, the allogenous TSPC-treated group exhibited increased collagen deposition and had better structural and biomechanical properties compared to the control group. This study thus demonstrated that the allogenous TSPC-seeded knitted silk–collagen sponge scaffold enhanced the efficacy of rotator cuff tendon regeneration by differentiating into tenocytes, and by secreting anti-inflammatory cytokines that prevent immunological rejection. Hence, allogenous TSPC-seeded knitted silk–collagen sponge scaffolds can be a clinically useful application for tendon tissue engineering.

Key words: Rotator cuff; Tissue engineering; Knitted silk–collagen sponge scaffold; Tendon stem/progenitor cell; Regeneration

1These authors provided equal contribution to this work.
Address correspondence to Hong Wei Ouyang, Mailbox #39, Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, 388 Yu Hang Tang Road, Hangzhou 310058, China. Tel/Fax: +86-571-88208262; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Weishan Chen, Department of Orthopedic Surgery, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, 88 Jie Fang Road, Hangzhou 310009, China. Tel: +86-571-87783545; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp.959 970–, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627516
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Inhibition of Collagen Deposit in Obstructed Rat Bladder Outlet by Transplantation of Superparamagnetic Iron Oxide-Labeled Human Mesenchymal Stem Cells as Monitored by Molecular Magnetic Resonance Imaging (MRI)

Hong Jun Lee,*†1 Jong Ho Won,‡1 Seung Hwan Doo,§ Jung Hoon Kim,¶ Ki Young Song,# Sun Ju Lee,** Inja Lim,†† Kyu-Tae Chang,‡‡ Yun Seob Song,§ and Seung U. Kim*†

*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
‡Department of Internal Medicine, Soonchunhyang University School of Medicine, Seoul, Korea
§Department of Urology, Soonchunhyang University School of Medicine, Seoul, Korea
¶Department of Radiology, Soonchunhyang University School of Medicine, Seoul, Korea
#Avon Old Farms School, Avon, CT, USA
**Kyunghee University School of Medicine, Seoul, Korea
††Department of Physiology, Chung-Ang University College of Medicine, Seoul, Korea
‡‡National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Korea

Bladder outlet obstruction (BOO) caused by collagen deposit is one of the most common problems in elderly males. The present study is to investigate if human mesenchymal stem cells (MSCs) are capable of inhibiting collagen deposition and improve cystometric parameters in bladder outlet obstruction in rats. Human MSCs were labeled with nanoparticles containing superparamagnetic iron oxide (SPION), and transplanted in rat BOO lesion site. Forty 6-week-old female Sprague-Dawley rats were divided into four groups (group 1: control, group 2: sham operation, group 3: BOO, and group 4: BOO rats receiving SPION-hMSCs). Two weeks after the onset of BOO, 1 × 106 SPION-hMSCs were injected into the bladder wall. Serial T2-weighted MR images were taken immediately after transplantation of SPION-labeled human MSCs and at 4 weeks posttransplantation. T2-weighted MR images showed a clear hypointense signal induced by the SPION-labeled MSCs. While the expression of collagen and TGF-β protein increased after BOO, the expression of both returned to the original levels after MSC transplantation. Expression of HGF and c-met protein also increased in the group with MSC transplantation. Maximal voiding pressure and residual urine volume increased after BOO but they recovered after MSC transplantation. Human MSCs transplanted in rat BOO models inhibited the bladder fibrosis and mediated recovery of bladder dysfunction. Transplantation of MSC-based cell therapy could be a novel therapeutic strategy against bladder fibrosis in patients with bladder outlet obstruction.

Key words: Bladder outlet obstruction; Human mesenchymal stem cells (MSCs); Collagen; Transforming growth factor-β (TGF-β); Superparamagnetic iron oxide nanoparticle (SPION); Magnetic resonance imaging (MRI); Cell transplantation

1These authors provided equal contribution to this work.
Address correspondence to Seung U. Kim, M.D., Ph.D., Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, BC V6T2B5, Canada. Tel: 604-822-7145; Fax: 604-822-7897; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Yun Seob Song, M.D., Ph.D., Department of Urology, Soonchunhyang University School of Medicine, Seoul 140-743, Korea. Tel: 82-2-709-9114; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 971–984, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627525
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Autologous Mesenchymal Stem Cells Prevent Transplant Arteriosclerosis by Enhancing Local Expression of Interleukin-10, Interferon-γ, and Indoleamine 2,3-dioxygenase

Hsiang-Yiang Jui,*1 Cheng-Hsin Lin,†1 Wan-Tseng Hsu,* Yi-Ru Liu,‡ Ron-Bin Hsu,§ Bor-Luen Chiang,† Wen-Yih I. Tseng,¶ Ming-Fong Chen,* Kenneth K. Wu,‡ and Chii-Ming Lee*‡

*Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan
†Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan
‡Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan
§Department of Surgery, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan
¶Center for Optoelectronic Biomedicine, National Taiwan University College of Medicine, Taipei, Taiwan

Transplant arteriosclerosis (TA) remains the major limitation of long-term graft survival in heart transplantation despite the advances in immunosuppressants. Mesenchymal stem cells (MSCs) have been demonstrated to suppress allogeneic immune responses by numerous in vitro studies. However, the immunomodulatory effects of MSCs in vivo are controversial and the underlying molecular mechanisms are not conclusive. In this study, we investigated the therapeutic potential of autologous bone marrow-derived MSCs on TA in a porcine model of femoral artery transplantation. MSCs or saline were injected into the soft tissue surrounding the arterial grafts immediately postanastomosis. Four weeks after transplantation, neointimal formation increased significantly in untreated allografts compared with the MSC-treated grafts as assessed by intravascular ultrasound (maximum luminal area stenosis: 40 ± 12% vs. 18 ± 6%, p < 0.001). Grafts harvested at 4 weeks showed dense perivascular lymphocyte infiltration accompanied by significant intimal hyperplasia in the untreated but not in the MSC-treated allografts. Serial angiographic examination showed that all of the untreated allografts became occluded at the 8th week whereas the majority of the MSC-treated grafts remained patent at the 12th week posttransplantation (n = 12 each group, p < 0.001). Quantitative PCR analysis revealed that Foxp3 expression was comparable between the untreated and the MSC-treated groups. However, expression of interleukin-10 (IL-10), interferon-γ (IFN-γ), and indoleamine 2,3-dioxygenase (IDO) was increased significantly in the MSC-treated allografts compared with that in the allograft controls (p = 0.021 for IL-10, p = 0.003 for IFN-γ, and p = 0.008 for IDO). In conclusion, local delivery of autologous MSCs alleviates TA by inducing allograft tolerance via enhanced expression of IL-10, IFN-γ, and IDO but not Foxp3-positive cells in the vessel wall. These results suggest that MSCs induce immune tolerance by activating the type 1 regulatory T-like cells.

Key words: Immunomodulation; Mesenchymal stem cells (MSCs); Transplant arteriosclerosis

1These authors provided equal contribution to this work.
Address correspondence to Chii-Ming Lee, M.D., Ph.D., Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan S. Road, Taipei, Taiwan. Tel: +886 972651087; Fax: +886 2 23934176; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 985–996, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X593145
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Trichostatin A Promotes Cardiomyocyte Differentiation of Rat Mesenchymal Stem Cells After 5-Azacytidine Induction or During Coculture With Neonatal Cardiomyocytes Via a Mechanism Independent of Histone Deacetylase Inhibition

Ge Yang,* Jie Tian,† Chuan Feng,* Li-li Zhao,* Zhenguo Liu,‡ and Jing Zhu*

*Department of Cell and Molecular Biology, Pediatric Institute of Chongqing Medical University, Chongqing, China
†Department of Cardiology, Children’s Hospital of Chongqing Medical University, Chongqing, China
‡Davis Heart & Lung Research Institute and Division of Cardiovascular Medicine, the Ohio State University Medical Center, Columbus, OH, USA

This study was to investigate the effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on cardiac differentiation of bone marrow mesenchymal stem cells (MSCs) in vitro. Rat MSCs were isolated and divided into six groups: 1) control; 2) 5-azacytidine treatment (5-aza, 10 μM); 3) treatment with TSA (100, 300, and 500 nM); 4) treatment with 5-aza followed by incubation with TSA; 5) coculture with neonatal cardiomyocytes (CMs); and 6) treatment with TSA then coculture with CMs. HDAC activity was significantly inhibited in TSA-treated cells with the maximal inhibition after 24 h of exposure to TSA at 300 nM. No changes in HDAC activity were observed in control, 5-aza-treated, or coculture groups. Following 7 days of differentiation, the expression of early cardiac transcription factors GATA-4, NKx2.5, MEF2c, and cardiac troponin T (cTnT) was increased by 6–8 times in the cells in 5-aza-treated, coculture, or TSA-treated groups over control as determined using real-time PCR, immunofluorescence staining, and Western blotting. However, the percent cTnT-positive cells were dramatically different with 0.7% for control, 10% for 5-aza-treated, 25% for coculture, and 4% for TSA-treated group (500 nM). TSA treatment of the cells pretreated with 5-aza or cocultured with CMs dramatically increased the expression of GATA-4, NKx2.5, and MEF2c by 35–50 times over control. The cTnT protein expression was also significantly increased by over threefold by TSA treatment (500 nM) in both 5-aza-treated and coculture group over control. The percent cTnT-positive cells in both 5-aza-pre-treated and coculture groups were significantly increased by TSA treatment after 1 week of differentiation by up to 92.6% (from 10.3% to 19.8%) and 23.9% (from 24.5% to 30.2%), respectively. These data suggested that TSA enhanced the cardiac differentiation of MSCs after 5-aza induction or during coculture with CMs through a mechanism beyond the inhibition of HDAC activity.

Key words: Mesenchymal stem cells; Cardiac differentiation; Trichostatin A; Histone deacetylase; 5-Azacytidine; Coculture

Address correspondence to Professor Jing Zhu, Department of Cell and Molecular Biology, The Children’s Hospital of Chongqing Medical University, 136 Zhongshan Er Road, Yu Zhong District, Chongqing 400014 P.R. China. Tel: 86-023-68486767; Fax: 86-023-68485111; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 997–1009, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X603611
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Multiple Intravenous Transplantations of Mesenchymal Stem Cells Effectively Restore Long-Term Blood Glucose Homeostasis by Hepatic Engraftment and β-Cell Differentiation in Streptozocin-Induced Diabetic Mice

Jennifer H. Ho,*†‡ Tzu-Ching Tseng,§ Wei-Hsien Ma,† Wei-Kee Ong,† Yu-Fan Chen,§ Ming-Hsiang Chen,§ Ming-Wei Lin,¶ Chuang-Ye Hong,*# and Oscar K. Lee§**††

*Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan
†Center for Stem Cell Research, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan
‡Department of Ophthalmology, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan
§Department of Medical Research & Education, Taipei Veterans General Hospital, Taipei, Taiwan
¶Institute of Public Health, National Yang-Ming University, Taipei, Taiwan
#Department of Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan
**Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
††Stem Cell Research Center, National Yang-Ming University, Taipei, Taiwan

Depletion of pancreatic β-cells results in insulin insufficiency and diabetes mellitus (DM). Single transplantation of mesenchymal stem cells exhibits short-term effects in some preclinical studies. Here, we further investigated the long-term therapeutic effects of multiple intravenous MSC transplantations. In this study, multiple human MSC transplantations (4.2 × 107 cells/kg each time) were performed intravenously at 2-week intervals into streptozocin (STZ)-induced diabetic mice for 6 months. Blood sugar, insulin, renal function, cholesterol, and triglyceride levels were monitored. We demonstrated that compared to single intravenous transplantation, which only transiently decreased hyperglycemia, multiple MSC transplantations effectively restored blood glucose homeostasis. Systemic oxidative stress levels were reduced from the seventh week of treatment. From the 11th week, production of human insulin was markedly increased. When MSC transplantation was skipped after blood sugar level returned to normal at the end of 15th week, a sharp rebound of blood sugar occurred, and was then controlled by subsequent transplantations. At the end of 6 months, histopathology examination revealed MSCs specifically engrafted into liver tissues of the recipients. Fifty-one percent of human cells in the recipient liver coexpressed human insulin, especially those surrounding the central veins. Taken together, intravenous MSC delivery was safe and effective for blood glucose stabilization in this preclinical DM model. Multiple transplantations were essential to restore and maintain glucose homeostasis through decreasing systemic oxidative stress in the early stage and insulin production in the late stage. Liver engraftment and differentiation into insulin-producing cells account for the long-term therapeutic effects of MSCs.

Key words: Mesenchymal stem cells (MSCs); Multiple intravenous transplantations; Blood glucose homeostasis; β-Cell depletion; Liver engraftment

Address correspondence to Oscar Kuang-Sheng Lee, MD, Ph.D., Department of Medical Research & Education, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei 112, Taiwan, R.O.C. Tel: 886 -2- 28712121 ext. 2640; Fax: 886 -2- 2875-7657; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1011–1021, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X623916
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Soluble Factors From Multipotent Mesenchymal Stromal Cells Have Antinecrotic Effect on Cardiomyocytes In Vitro and Improve Cardiac Function in Infarcted Rat Hearts

P. Fidelis-de-Oliveira,*† J. P. S. Werneck-de-Castro,*‡ V. Pinho-Ribeiro,* B. C. M. Shalom,* J. H. Nascimento-Silva,* R. H. Costa e Souza,* I. S. Cruz,* R. R. Rangel,* R. C. S. Goldenberg,* and A. C. Campos-de-Carvalho*§

*Instituto de Biofísica Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brazil
†Área de Ciências Fisiológicas, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Brazil
‡Departamento de Biociências da Atividade Física, EEFD-UFRJ, Rio de Janeiro, Brazil
§Albert Einstein College of Medicine, Bronx, NY, USA

The mechanisms underlying the functional improvement after injection of multipotent mesenchymal stromal cells (MSCs) in infarcted hearts remain incompletely understood. The aim of this study was to investigate if soluble factors secreted by MSCs promote cardioprotection. For this purpose, conditioned medium (CM) was obtained after three passages from MSC cultures submitted to 72 h of conditioning in serum-free DMEM under normoxia (NCM) or hypoxia (HCM) conditions. CM was concentrated 25-fold before use (NCM-25X, concentrated normoxia conditioned medium; HCM-25X, concentrated hypoxia conditioned medium). The in vitro cardioprotection was evaluated in neonatal ventricular cardiomyocytes by quantifying apoptosis after 24 h of serum deprivation associated with hypoxia (1% O2) in the absence or presence of NCM and HCM (nonconcentrated and 25-fold concentrated). The in vivo cardioprotection of HCM was tested in a model of myocardial infarction (MI) induced in Wistar male rats by permanent left coronary occlusion. Intramyocardial injection of HCM-25X (n = 14) or nonconditioned DMEM (n = 16) was performed 3 h after coronary occlusion and cardiac function was evaluated 19–21 days after medium injection. Cardiac function was evaluated by electro- and echocardiogram, left ventricular catheterization, and treadmill test. The in vitro results showed that HCM was able to decrease cardiomyocyte necrosis. The in vivo results showed that HCM-25X administered 3 h after AMI was able to promote a significant reduction (35%) in left ventricular end-diastolic pressure and improvement of cardiac contractility (15%) and relaxation (12%). These results suggest that soluble factors released in vitro by MSCs are able to promote cardioprotection in vitro and improve cardiac function in vivo.

Key words: Multipotent mesenchymal stromal cells (MSCs); Conditioned medium; Paracrine effect; Myocardial infarction; Apoptosis/necrosis

Address correspondence to Prof. Antônio Carlos Campos de Carvalho, Universidade Federal do Rio de Janeiro (UFRJ), Instituto de Biofísica Carlos Chagas Filho, CCS, bloco G, Ilha do Fundão, 21941-902, Rio de Janeiro, RJ, Brasil. Tel: +55-21/2562-6558; Fax: +55-21/2280-8193; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1023–1037, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X623862
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adipose Stromal Vascular Fraction Improves Cardiac Function in Chronic Myocardial Infarction Through Differentiation and Paracrine Activity

Manuel Mazo,* Arantxa Cemborain,* Juan Jose´ Gavira,† Gloria Abizanda,* Miriam Araña,* Mayte Casado,‡ Mario Soriano,‡ Salomón Hernández,* Cristina Moreno,§ Margarita Ecay,¶ Edurne Albiasu,* Miriam Belzunce,# Josune Orbe,# José Antonio Páramo,# Juana Merino,‡ Iván Peñuelas,§ José Manuel García Verdugo,‡ Beatriz Pelacho,* and Felipe Prosper*

*Hematology and Cell Therapy and Foundation for Applied Medical Research, Division of Cancer, Clínica Universitaria, University of Navarra, Navarra, Spain
†Department of Cardiology and Cardiovascular Surgery, Clínica Universitaria, University of Navarra, Navarra, Spain
‡Department of Cell Biology, Instituto Cavanilles, University of Valencia, Valencia, Spain
§Immunology Service, Clínica Universitaria, University of Navarra, Navarra, Spain
¶Department of Nuclear Medicine and MicroPET Research Unit CIMA-CUN, Clínica Universitaria, University of Navarra, Navarra, Spain
#Foundation for Applied Medical Research, Division of Cardiovascular Diseases, Navarra, Spain

Fresh adipose-derived cells have been shown to be effective in the treatment of acute myocardial infarction (MI), but their role in the chronic setting is unknown. We sought to determine the long-term effect of the adipose derived-stromal vascular fraction (SVF) cell transplantation in a rat model of chronic MI. MI was induced in 82 rats by permanent coronary artery ligation and 5 weeks later rats were allocated to receive an intramyocardial injection of 107 GFP-expressing fresh SVF cells or culture media as control. Heart function and tissue metabolism were determined by echocardiography and 18F-FDG-microPET, respectively, and histological studies were performed for up to 3 months after transplantation. SVF induced a statistically significant long-lasting (3 months) improvement in cardiac function and tissue metabolism that was associated with increased revascularization and positive heart remodeling, with a significantly smaller infarct size, thicker infarct wall, lower scar fibrosis, and lower cardiac hypertrophy. Importantly, injected cells engrafted and were detected in the treated hearts for at least 3 months, directly contributing to the vasculature and myofibroblasts and at negligible levels to cardiomyocytes. Furthermore, SVF release of angiogenic (VEGF and HGF) and proinflammatory (MCP-1) cytokines, as well as TIMP1 and TIMP4, was demonstrated in vitro and in vivo, strongly suggesting that they have a trophic effect. These results show the potential of SVF to contribute to the regeneration of ischemic tissue and to provide a long-term functional benefit in a rat model of chronic MI, by both direct and indirect mechanisms.

Key words: Chronic myocardial infarction (MI); Stromal vascular fraction (SVF); Cardiac function

Address correspondence to Beatriz Pelacho, Ph.D., Foundation for Applied Medical Research, Av. Pío XII 57, Pamplona 31008, Navarra, Spain. Tel: 34 948 194700; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Felipe Prósper, M.D., Hematology and Cell Therapy, Clínica Universitaria, Av. Pío XII 36, Pamplona 31008, Navarra, Spain. Tel: 34 948 255400; Fax: 34 948 296500; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1039–1053, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X601000
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Inhibiting Matrix Metalloproteinase by Cell-Based Timp-3 Gene Transfer Effectively Treats Acute and Chronic Ischemic Cardiomyopathy

Hai Tian,*†1 Ming-Li Huang,†‡1 Kai-Yu Liu,*† Zhi-Bo Jia,*† Lu Sun,*† Shu-Lin Jiang,*† Wei Liu,† Heather Y. McDonald Kinkaid,§ Jun Wu,§ and Ren-Ke Li§

*Department of Cardiovascular Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, China
†Key Laboratories of Myocardial Ischemia Mechanism and Treatment, Harbin Medical University, Ministry of Education, Harbin, China
‡Department of Gynecology and Obstetrics, First Clinical College of Harbin Medical University, Harbin, China
§Department of Surgery, Division of Cardiovascular Surgery, University of Toronto and Toronto General Research Institute, University Health Network, Toronto, Ontario, Canada

After a myocardial infarction (MI), an increase in the cardiac ratio of matrix metalloproteinases (MMPs) relative to their inhibitors (TIMPs) causes extracellular matrix modulation that leads to ventricular dilatation and congestive heart failure. Cell therapy can mitigate these effects. In this study, we tested whether increasing MMP inhibition via cell-based gene transfer of Timp-3 further preserved ventricular morphometry and cardiac function in a rat model of MI. We also measured the effect of treatment timing. We generated MI (coronary artery ligation) in adult rats. Three or 14 days later, we implanted medium (control) or vascular smooth muscle cells transfected with empty vector (VSMCs) or Timp-3 (C-TIMP-3) into the peri-infarct region (n = 15–24/group). We assessed MMP-2 and -9 expression and activity, TIMP-3, and TNF-α expression, cell apoptosis, infarct size and thickness, ventricular morphometry, and cardiac function (by chocardiography). Relative to medium, VSMCs delivered at either time point significantly reduced cardiac expression and activity of MMP-2 and -9, reduced expression of TNF-α, and increased expression of TIMP-3. Cell therapy also reduced apoptosis and scar area, increased infarct thickness, preserved ventricular structure, and reduced functional loss. All these effects were augmented by C-TIMP-3 treatment. Survival and cardiac function were significantly greater when VSMCs or C-TIMP-3 were delivered at 3 (vs. 14) days after MI. Upregulating post-MI cardiac TIMP-3 expression via cell-based gene therapy contributed additional regulation of MMP, TIMP, and TNF-α levels, thereby boosting the structural and functional effects of VSMCs transplanted at 3 or 14 days after an MI in rats. Early treatment may be superior to late, though both are effective.

Key words: Cell-based gene therapy; Extracellular matrix remodeling; Myocardial infarction (MI); Tissue inhibitors of matrix metalloproteinases (TIMPs)

1These authors provided equal contribution to this work.
Address correspondence to Ren-Ke Li, M.D., Ph.D., MaRS Centre, Toronto Medical Discovery Tower, Room #3-702, 101 College St., Toronto, ON, Canada M5G 1L7. Tel: 416-581-7492; Fax: 416-581-7493; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Hai Tian, M.D., Ph.D., Department of Cardiovascular Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150086 China. Tel: 86-451-86605248; Fax: 86-451-82569696; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1055–1061, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627499
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Brief Communication

Cotreatment With Darbepoetin and Granulocyte Colony-Stimulating Factor Is Efficient to Recruit Proangiogenic Cell Populations in Patients With Acute Myocardial Infarction

Hyun-Jae Kang,*† Eun-Jung Yoon,† Eun-Ju Lee,† Min-Kyung Kim,* Jung-Won Suh,‡ Kyung-Woo Park,* Hae-Young Lee,* Kyoung Un Park,§ Young-Seok Cho,‡ Bon-Kwon Koo,* In-Ho Chae,‡ Dong-Ju Choi,‡ Kyu-Sup Han,¶ Hyo-Soo Kim,*† and Young-Bae Park*†

*Department of Internal Medicine, Seoul National University Hospital, Seoul, Korea
†Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea
‡Cardiovascular Center, Seoul National University Bundang Hospital, Seongnam, Korea
§Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea
¶Department of Laboratory Medicine, Seoul National University Hospital, Seoul, Korea

To determine whether newer combination cytokine treatment with granulocyte colony-stimulating factor (GCSF) and darbepoetin can improve efficacy of stem cell therapy, we evaluated safety and peripheral blood stem/progenitor cell (PBSC) mobilizing effects of combination cytokine in comparison with G-CSF alone in patients with acute myocardial infarction (AMI). We randomized 60 patients with AMI into two groups under 2:1 ratio; combination treatment with darbepoetin and G-CSF (n = 41: Combicytokine group) and the G-CSF alone (n = 19: G-CSF group). After coronary angioplasty, G-CSF was treated for 3 days with dose of 10 μg/kg/day in both groups. Only in the combicytokine group, additional single intravenous injection of 4.5 μg/kg of darbepoetin was administrated immediate after coronary angioplasty. Combination cytokine treatment was well tolerated as was G-CSF alone. PBSCs were obtained by apheresis for intracoronary infusion after completion of cytokine treatment and were analyzed by flow cytometry. The purity of proangiogenic cells was higher in combination cytokine group than the G-CSF group. Specifically, proportion of CD34+/KDR+ endothelial progenitor cells, CD3+/CD31+ angiogenic T cells and Tie2+/CXCR4+ cells in apheresis products were higher in the combicytokine group. These meant that the combicytokine treatment recruited PBSCs in higher purity and fewer unwanted inflammatory cells than G-CSF alone in apheresis products. Combination treatment with darbepoetin and G-CSF is safe and more efficient to mobilize and recruit proangiogenic cells than G-CSF alone in patients with AMI. (Trial registration: www.ClinicalTrials.gov identifier: NCT00501917)

Key words: Granulocyte colony-stimulating factor (G-CSF); Erythropoietin; Acute myocardial infarction (AMI); Stem cell; Progenitor cell

Address correspondence to Hyo-Soo Kim, Department of Internal Medicine, Seoul National University Hospital, 28 Yongon-dong Chongno-gu, Seoul, 110-744 Korea. Tel: 82-2-2072-2226; Fax: 82-2-766-8904; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it