Cell Transplantation 22(8) Abstracts

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Cell Transplantation, Vol. 22, pp. 1309-1323, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657260
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Stem Cell-Based Therapy for Spinal Cord Injury

Vladislav Volarevic,* Slaven Erceg,†‡ Shom Shanker Bhattacharya,† Petra Stojkovic,*§ Philip Horner,¶ and Miodrag Stojkovic*§

*Center for Molecular Medicine and Stem Cell Research, Medical Faculty, University of Kragujevac, Serbia
†CABIMER, Seville, Spain
‡Medical Genome Project, Seville, Spain
§SPEBO MEDICAL, Leskovac, Serbia
¶Program in Neurobiology and Behavior, University of Washington School of Medicine and Institute for Stem Cell and Regenerative Medicine, Seattle, WA, USA

Stem cells (SCs) represent a new therapeutic approach for spinal cord injury (SCI) by enabling improved sensory and motor functions in animal models. The main goal of SC-based therapy for SCI is the replacement of neurons and glial cells that undergo cell death soon after injury. Stem cells are able to promote remyelination via oligodendroglia cell replacement to produce trophic factors enhancing neurite outgrowth, axonal elongation, and fiber density and to activate resident or transplanted progenitor cells across the lesion cavity. While several SC transplantation strategies have shown promising yet partial efficacy, mechanistic proof is generally lacking and is arguably the largest impediment toward faster progress and clinical application. The main challenge ahead is to spur on cooperation between clinicians, researchers, and patients in order to define and optimize the mechanisms of SC function and to establish the ideal source/s of SCs that produce efficient and also safe therapeutic approaches.

Key words: Stem cells (SCs); Spinal cord injury (SCI); Differentiation; Remyelination; Inflammation

Received January 5, 2012; final acceptance May 23, 2012. Online prepub date: October 3, 2012.
Address correspondence to Miodrag Stojkovic, Center for Molecular Medicine and Stem Cell Research, Medical Faculty, University of Kragujevac, Serbia. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1325-1336, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X659899
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Review

Endothelial Progenitor Cell-Based Therapy for Pulmonary Arterial Hypertension

Jin-Xiu Yang,* Yan-Yun Pan,* You-Yang Zhao,† and Xing-Xiang Wang*

*Department of Cardiology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, PR China
†Department of Pharmacology and Center for Lung and Vascular Biology, University of Illinois College of Medicine, Chicago, IL, USA

A growing body of evidence in animal models and clinical studies supports the concept that endothelial progenitor cell (EPC)-mediated therapy ameliorates pulmonary arterial hypertension (PAH) and thus may represent a novel approach to treat it. Conversely, several experimental findings suggest that EPCs may be involved in PAH pathogenesis and disease progression. These discrepant results confuse the application of EPC transplantation as an effective treatment strategy for PAH. To improve the study of EPC transplantation in PAH therapy, it is high time that we resolve this dilemma. In this review, we examine the pathobiological changes of PAH, the characteristics of EPCs, and the underlying mechanisms of EPC effects on PAH.

Key words: Pulmonary arterial hypertension (PAH); Endothelial progenitor cells (EPCs); Transplantation; Mechanisms

Received August 11, 2011; final acceptance July 31, 2012. Online prepub date: January 2, 2013.
Address correspondence to Prof. Xing-Xiang Wang, No. 79 Qingchun Road, Hangzhou, Zhejiang Province, PR China, 310003. Tel: +86-571-87236500; Fax: +86-571-87236889; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1337-1349, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657657
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Human Amnion Epithelial Cells Repair Established Lung Injury

Patricia Vosdoganes,*† Euan M. Wallace,*† Siow Teng Chan,* Rutu Acharya,* Tim J. M. Moss,*† and Rebecca Lim*

*The Ritchie Centre, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia
†Department of Obstetrics and Gynecology, Monash Medical Centre, Monash University, Clayton, Victoria, Australia

With a view to developing a cell therapy for chronic lung disease, human amnion epithelial cells (hAECs) have been shown to prevent acute lung injury. Whether they can repair established lung disease is unknown. We aimed to assess whether hAECs can repair existing lung damage induced in mice by bleomycin and whether the timing of cell administration influences reparative efficacy. In addition, we aimed to characterize the effect of hAECs on fibroblast proliferation and activation, investigating possible mechanisms of reparative action. hAECs were administered intraperitoneally (IP) either 7 or 14 days after bleomycin exposure. Lungs were assessed 7 days after hAEC administration. Bleomycin significantly reduced body weight and induced pulmonary inflammation and fibrosis at 14 and 21 days. Delivery of hAECs 7 days after bleomycin had no effect on lung injury, whereas delivery of hAECs 14 days after bleomycin normalized lung tissue density, collagen content, and α-SMA production, in association with a reduction in pulmonary leucocytes and lung expression of TGF-β, PDGF-α, and PDGF-β. In vitro, hAECs reduced proliferation and activation of primary mouse lung fibroblasts. Our findings suggest that the timing of hAEC administration in the course of lung disease may impact on the ability of hAECs to repair lung injury.

Key words: Amnion epithelial cells; Bleomycin; Fibrosis; Inflammation; Chronic lung disease; Regenerative medicine

Received October 22, 2011; final acceptance August 10, 2012. Online prepub date: October 4, 2012.
Address correspondence to Dr. Rebecca Lim, The Ritchie Centre, Monash Institute of Medical Research, Level 3, 27-31 Wright Street, Clayton, Victoria 3168, Australia. Tel: +61 3 9594 7410; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1351-1367, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657387
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Autocrine TGF-β Induces Epithelial to Mesenchymal Transition in Human Amniotic Epithelial Cells

Antonia Alcaraz,*1 Anna Mrowiec,*1 Carmen L. Insausti,† Eva M. García-Vizcaíno,* Catalina Ruiz-Canada,* María C. López-Martínez,* José M. Moraleda,† and Francisco J. Nicolás*

*Oncología Molecular y TGF-β, Unidad de Investigación, Hospital Universitario Virgen de la Arrixaca, El Palmar, Murcia, Spain
†Unidad de Terapia Celular, Hospital Universitario Virgen de la Arrixaca, El Palmar, Murcia, Spain

Human amniotic epithelial cells (hAECs) have been the object of intense research due to their potential therapeutic use. In this paper, we present molecular evidence of a bona fide epithelial to mesenchymal transition (EMT) undergone by hAECs. Amniotic membrane (AM)-derived hAECs showed the presence of typical epithelial markers such as E-cadherin and cytokeratins. hAECs in culture, however, underwent morphological changes acquiring a mesenchymal shape. Epithelial cell markers were lost and typical mesenchymal markers, such as vimentin and a-SMA, appeared. Several genes associated with EMT, such as SNAI1, MMP9, PAI1, or ACTA2, increased their expression. The expression of the transcription activators KLF4 or MTA3 was consistent with the downregulation of CDH1. We have shown that hAECs undergo EMT due to the autocrine production of TGF-β. Furthermore, the addition of the TGF-β receptor I (ALK5) inhibitor SB-431542 or TGF-β neutralizing antibody to hAECs prevented EMT and preserved the hAECs’ epithelial phenotype. Altogether, these results suggest that cultured hAECs undergo EMT through the autocrine production of TGF-β.

Key words: Amniotic membrane (AM); Epithelial to mesenchymal transition (EMT); Human amniotic epithelial cells (hAECs); Transforming growth factor-β (TGF-β)

Received November 23, 2011; final acceptance July 7, 2012. Online prepub date: October 2, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Francisco J. Nicolás, Laboratorio de Oncología Molecular y TGF-β, Hospital Universitario Virgen de la Arrixaca, Crta Madrid-Cartagena, s/n, 30120 El Palmar, Murcia, Spain. Tel: +34 968 369036; Fax: +34 968 369438; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1369-1380, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X659853
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Adult Human Liver Mesenchymal Stem/Progenitor Cells Participate in Mouse Liver Regeneration After Hepatectomy

Dung Ngoc Khuu, Omar Nyabi, Cédric Maerckx, Etienne Sokal, and Mustapha Najimi

Université Catholique de Louvain, Institut de Recherche Clinique et Expérimentale (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Brussels, Belgium

The advances in stem cell science have promoted research on their use in liver regenerative medicine. Beyond the demonstration of their ability to display metabolic functions in vitro, candidate cells should demonstrate achievable in situ differentiation and ability to participate to liver repopulation. In this work, we studied the in vivo behavior of adult liver mesenchymal stem/progenitor cells (ADHLSCs) after transplantation into immunodeficient mice. The kinetics of engraftment and in situ hepatogenic differentiation were analyzed. Response of transplanted ADHLSCs to regenerative stimulus was also evaluated. Nondifferentiated ADHLSCs were intrasplenically transplanted into SCID mice. Efficiency of transplantation was evaluated at the level of engraftment and in situ differentiation using immunohistochemistry, in situ hybridization, and RT-PCR. After bromodeoxyuridine (BrdU) implantation, proliferation of transplanted ADHLSCs in response to 20% hepatectomy was assessed using immunohistochemistry. We demonstrated that ADHLSC engraftment in the SCID mouse liver was low but remained stable up to 60 days posttransplantation, when albumin (ALB) immunopositive ADHLSCs were still detected and organized as clusters. Coexpression of ornithine transcarbamylase (OTC) demonstrated ADHLSC in situ differentiation mostly near the hepatic portal vein. After 20% hepatectomy on 1 month transplanted mice, the percentage of BrdU and human ALB immunopositive ADHLSCs increased from 3 to 28 days post-BrdU implantation to reach 31.3 ± 5.4% of the total analyzed human cells. In the current study, we demonstrate that transplanted ADHLSCs are able to differentiate in the non preconditioned SCID mouse liver mainly in the periportal area. In response to partial hepatectomy, integrated ADHLSCs proliferate and participate to recipient mouse liver regeneration.

Key words: Stem/progenitor cells; Liver regeneration; Liver; Hepatocyte; Engraftment; In situ differentiation

Received June 22, 2011; final acceptance August 15, 2012. Online prepub date: December 4, 2012.
Address correspondence to Mustapha Najimi, Ph.D., Laboratory of Pediatric Hepatology and Cell Therapy, Avenue Hippocrate 10/11, 1200, Brussels, Belgium. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1381-1394, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657666
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Reduced Efficacy of Mesenchymal Stromal Cells in Preventing Graft-Versus-Host Disease in an In Vivo Model of Haploidentical Bone Marrow Transplant With Leukemia

Alberto Oviedo,* Rosa Yañez,* Isabel Colmenero,† Montserrat Aldea,* Antonio Rubio,* Juan Antonio Bueren,* and María Luisa Lamana*

*Hematopoiesis and Gene Therapy Division, Centro de Investigaciones Energéticas Medioambientales y Tecnológicas (CIEMAT) and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBER-ER), Madrid, Spain
†Department of Pathology, Hospital Niño Jesús, Madrid, Spain

Mesenchymal stromal cell (MSC) immunosuppressive properties have been applied to treat graft-versus-host disease (GVHD) in allogeneic hematopoietic stem cell transplants (HSCTs). We have previously demonstrated that MSC infusions early after haplo-HSCT prevent GVHD in a haploidentical-HSCT mouse model. Now, we investigated the impact that MSCs’ immunosuppressive properties have on the graft-versus-leukemia (GVL) effect. First, to mimic a chronic myeloid leukemia (CML) relapse after a haploidentical HSCT, lethally irradiated mice were coinfused with haploidentical donor bone marrow cells plus syngenic hematopoietic progenitors transduced with a retroviral vector encoding both the BCR/ABL oncogene and the DNGFR marker gene. As expected, a CML-like myeloproliferative syndrome developed in all the recipient animals. The addition of haploidentical splenocytes to the transplanted graft prevented CML development by a GVL effect, and all transplanted recipients died of GVHD. This GVL mouse model allowed us to investigate the impact of MSCs infused to prevent GVHD on days 0, 7, and 14 after HSCT, on the GVL effect, expecting an increase in leukemic relapse. Strikingly, a high mortality of the recipients was observed, caused by GVHD, and only few leukemic cells were detected in the recipient animals. In contrast, GVHD prevention by MSCs in the absence of BCR/ABL leukemic cells resulted in a significant survival of the recipients. In vitro data pointed to an inability of MSCs to control strong CTLs responses against BCR/ABL. Our results show that, although an evident increase in leukemic relapses induced by MSCs could not be detected, they showed a reduced efficacy in preventing GVHD that precluded us to draw clear conclusions on MSCs’ impact over GVL effect.

Key words: Mesenchymal stromal cells (MSCs); Graft-versus-leukemia (GVL) effect; Graft-versus-host disease (GVHD) prevention; Mouse model

Received July 1, 2011; final acceptance July 25, 2012. Online prepub date: October 4, 2012.
Address correspondence to María Luisa Lamana, Hematopoiesis and Gene Therapy Division, CIEMAT/CIBER-ER Avenida Complutense, No. 22, 28040 Madrid, Spain. Tel: +34-913460890; Fax: +34-913466484; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1395-1408, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653264
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Intra-articular Delivery of Purified Mesenchymal Stem Cells From C57BL/6 or MRL/MpJ Superhealer Mice Prevents Posttraumatic Arthritis


Brian O. Diekman,*† Chia-Lung Wu,*† Craig R. Louer,* Bridgette D. Furman,* Janet L. Huebner,‡ Virginia B. Kraus,‡ Steven A. Olson,* and Farshid Guilak*†

*Department of Orthopaedic Surgery, Duke University Medical Center, Durham, NC, USA
†Department of Biomedical Engineering, Duke University, Durham, NC, USA
‡Department of Medicine, Duke University Medical Center, Durham, NC, USA

Joint injury dramatically enhances the onset of osteoarthritis (OA) and is responsible for an estimated 12% of OA. Posttraumatic arthritis (PTA) is especially common after intra-articular fracture, and no disease-modifying therapies are currently available. We hypothesized that the delivery of mesenchymal stem cells (MSCs) would prevent PTA by altering the balance of inflammation and regeneration after fracture of the mouse knee. Additionally, we examined the hypothesis that MSCs from the MRL/MpJ (MRL) “superhealer” mouse strain would show increased multilineage and therapeutic potentials as compared to those from C57BL/6 (B6) mice, as MRL mice have shown exceptional in vivo regenerative abilities. A highly purified population of MSCs was prospectively isolated from bone marrow using cell surface markers (CD45/TER119/PDGFRa+/Sca-1+). B6 MSCs expanded greater than 100,000-fold in 3 weeks when cultured at 2% oxygen and displayed greater adipogenic, osteogenic, and chondrogenic differentiation as compared to MRL MSCs. Mice receiving only a control saline injection after fracture demonstrated PTA after 8 weeks, but the delivery of 10,000 B6 or MRL MSCs to the joint prevented the development of PTA. Cytokine levels in serum and synovial fluid were affected by treatment with stem cells, including elevated systemic interleukin-10 at several time points. The delivery of MSCs did not reduce the degree of synovial inflammation but did show increased bone volume during repair. This study provides evidence that intra-articular stem cell therapy can prevent the development of PTA after fracture and has implications for possible clinical interventions after joint injury before evidence of significant OA.

Key words: Mesenchymal stem cells (MSCs); Osteoarthritis (OA); Posttraumatic arthritis (PTA); Intra-articular fracture; Cell therapy; Immunomodulation

Received March 5, 2012; final acceptance July 14, 2012. Online prepub date: August 10, 2012.
Address correspondence to Farshid Guilak, 375 Medical Sciences Research Bldg., Box 3093, Duke University Medical Center, Durham, NC 27710, USA. Tel: +1-919-684-2521; Fax: +1-919-681-8490; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1409-1425, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657620
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Distinct Immunomodulatory and Migratory Mechanisms Underpin the Therapeutic Potential of Human Mesenchymal Stem Cells in Autoimmune Demyelination

Natalie L. Payne,* Guizhi Sun,* Courtney McDonald,* Daniel Layton,* Leon Moussa,* Ashley Emerson-Webber,* Nadege Veron,* Christopher Siatskas,* Daniella Herszfeld,* John Price,† and Claude C. A. Bernard*

*Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Victoria, Australia
†Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia

Mesenchymal stem cells (MSCs) are efficacious in a variety of intractable diseases. While bone marrow (BM)-derived MSCs (BM-MSCs) have been widely investigated, MSCs from other tissue sources have also been shown to be effective in several autoimmune and inflammatory disorders. In the present study, we simultaneously assessed the therapeutic efficacy of human BM-MSCs, as well as MSCs isolated from adipose tissue (Ad-MSCs) and umbilical cord Wharton’s jelly (UC-MSCs), in experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis (MS). Prior to in vivo experiments, we characterized the phenotype and function of all three MSC types. We show that BM-MSCs were more efficient at suppressing the in vitro proliferation of mitogen or antigen-stimulated T-cell responses compared to Ad-MSCs and UC-MSCs. Notably BM-MSCs induced the differential expression of cytokines from normal and stimulated T-cells. Paradoxically, intravenous transplantation of BM-MSCs into C57Bl/6 mice with chronic progressive EAE had a negligible effect on the disease course, even when multiple MSC injections were administered over a number of time points. In contrast, Ad-MSCs had the most significant impact on clinical and pathological disease outcomes in chronic progressive and relapsing–remitting EAE models. In vivo tracking studies revealed that Ad-MSCs were able to migrate to the central nervous system (CNS), a property that most likely correlated with their broader expression of homing molecules, while BM-MSCs were not detected in this anatomic region. Collectively, this comparative investigation demonstrates that transplanted Ad-MSCs play a significant role in tissue repair processes by virtue of their ability to suppress inflammation coupled with their enhanced ability to home to the injured CNS. Given the access and relatively ease for harvesting adipose tissue, these data further implicate Ad-MSCs as a cell therapeutic that may be used to treat MS patients.

Key words: Multiple sclerosis (MS); Experimental autoimmune encephalomyelitis (EAE); Mesenchymal stem cells (MSCs); Immunomodulation; Cell migration

Received October 13, 2011; final acceptance July 28, 2012. Online prepub date: October 4, 2012.
Address correspondence to Prof. Claude C. A. Bernard, Monash Immunology and Stem Laboratories, Level 3 STRIP Building 75, Monash University, Wellington Road, Clayton, Victoria 3800, Australia. Tel: +61 3 9905 0623; Fax: +61 3 9905 0680; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1427-1440, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657314
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Therapeutic Potential of Human Induced Pluripotent Stem Cells in Experimental Stroke

Da-Jeong Chang,*1 Nayeon Lee,*1 In-Hyun Park,†‡1 Chunggab Choi,* Iksoo Jeon,* Jihye Kwon,* Seung-Hun Oh,* Dong Ah Shin,§ Jeong Tae Do,* Dong Ryul Lee,* Hyunseung Lee,¶ Hyeyoung Moon,¶ Kwan Soo Hong,¶ George Q. Daley,† and Jihwan Song*

*CHA Stem Cell Institute, Department of Biomedical Science, CHA University, Seoul, Korea
†Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA
‡Department of Genetics, Yale University School of Medicine, New Haven, CT, USA
§Department of Neurosurgery, Yonsei University College of Medicine, Seoul, Korea
¶Division of Magnetic Resonance, Korea Basic Science Institute, Ochang, Korea

Ischemic stroke mainly caused by middle cerebral artery occlusion (MCAo) is a major type of stroke, but there are currently very limited therapeutic options for its cure. Neural stem cells (NSCs) or neural precursor cells (NPCs) derived from various sources are known to survive and improve neurological functions when they are engrafted in animal models of stroke. Induced pluripotent stem cells (iPSCs) generated from somatic cells of patients are novel cells that promise the autologous cell therapy for stroke. In this study, we successfully differentiated iPSCs derived from human fibroblasts into NPCs and found their robust therapeutic potential in a rodent MCAo stroke model. We observed the significant graft-induced behavioral recovery, as well as extensive neural tissue formation. Animal MRI results indicated that the majority of contralaterally transplanted iPSC-derived NPCs migrated to the peri-infarct area, showing a pathotropism critical for tissue recovery. The transplanted animals exhibited the significant reduction of stroke-induced inflammatory response, gliosis and apoptosis, and the contribution to the endogenous neurogenesis. Our results demonstrate that iPSC-derived NPCs are effective cells for the treatment of stroke.

Key words: Stroke; Induced pluripotent stem cells (iPSCs); Behavioral recovery; Endogenous neurogenesis

Received March 3, 2012; final acceptance June 20, 2012. Online prepub date: October 3, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Jihwan Song, D.Phil., CHA Stem Cell Institute, Department of Biomedical Science, CHA University, 606-16 Yeoksam 1-dong, Kangnam-gu, Seoul 135-081, Korea. Tel: +82 2 3468 3393; Fax: +82 2 538 4102; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1441-1452, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657323
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Therapeutic Effect of BDNF-Overexpressing Human Neural Stem Cells (HB1.F3.BDNF) in a Rodent Model of Middle Cerebral Artery Occlusion

Da-Jeong Chang,* Nayeon Lee,* Chunggab Choi,* Iksoo Jeon,* Seung-Hun Oh,* Dong Ah Shin,† Tae-Sun Hwang,* Hong J. Lee,‡ Seung U. Kim,‡ Hyeyoung Moon,§ Kwan Soo Hong,§ Kyung-Sun Kang,¶ and Jihwan Song*

*CHA Stem Cell Institute, Department of Biomedical Science, CHA University, Seoul, Korea
†Department of Neurosurgery, Yonsei University College of Medicine, Seoul, Korea
‡Medical Research Institute, Chung-Ang University College of Medicine, Seoul, Korea
§Division of Magnetic Resonance, Korea Basic Science Institute, Ochang, Korea
¶Adult Stem Cell Research Center, College of Veterinary Medicine, Seoul National University, Seoul, Korea

Ischemic stroke mainly caused by middle cerebral artery occlusion (MCAo) represents the major type of stroke; however, there are still very limited therapeutic options for the stroke-damaged patients. In this study, we evaluated the neurogenic and therapeutic potentials of human neural stem cells (NSCs) overexpressing brain-derived neurotrophic factor (HB1.F3.BDNF) following transplantation into a rodent model of MCAo. F3.BDNF human NSCs (F3.BDNF) were transplanted into the contralateral side of striatum at 7 days after MCAo, and the transplanted animals were monitored up to 8 weeks using animal MRI and various behavioral tests before they were sacrificed for immunohistochemical analysis. Interestingly, animal MRI results indicate that the majority of contralaterally transplanted neural stem cells were migrated to the peri-infarct area, showing a pathotropism. Transplanted animals exhibited significant behavioral improvements in stepping, rotarod, and modified neurological severity score (mNSS) tests. We also found that the transplanted human cells were colocalized with nestin, DCX, MAP2, DARPP-32, TH, GAD65/67-positive cells, of which results can be correlated with neural regeneration and behavioral recovery in the transplanted animals. More importantly, we were able to detect high levels of human BDNF protein expression, presumably derived from the transplanted F3.BDNF. Taken together, these results provide strong evidence that human neural stem cells (F3.BDNF) are effective in treating stroke animal models.

Key words: Stroke; Neural stem cells (NSCs); Brain-derived neurotrophic factor (BDNF); Behavioral recovery; Animal MRI

Received March 3, 2012; final acceptance June 20, 2012. Online prepub date: December 3, 2012.
Address correspondence to Jihwan Song, D.Phil., CHA Stem Cell Institute, Department of Biomedical Science, CHA University, 606-16 Yeoksam 1-dong, Kangnam-gu, Seoul 135-081, Korea. Tel: +82 2 3468 3393; Fax: +82 2 538 4102; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1453-1462, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658755
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Embryonic Stem Cells Incorporate Into Newly Formed Bone and Do Not Form Tumors in an Immunocompetent Mouse Fracture Model

Jaymi T. Taiani,* Roman J. Krawetz,† Akihiro Yamashita,† Yves Pauchard,‡ Helen R. Buie,‡ Dragana Ponjevic,§ Steven K. Boyd,‡ Derrick E. Rancourt,† and John R. Matyas§

*McCaig Institute for Bone and Joint Health, Biomedical Engineering Program, Department of Medical Science, Faculty of Graduate Studies, University of Calgary, Calgary, Alberta, Canada
†McCaig Institute for Bone and Joint Health, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada
‡McCaig Institute for Bone and Joint Health, Schulich School of Engineering, University of Calgary, Calgary, Alberta, Canada
§McCaig Institute for Bone and Joint Health, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada

Embryonic stem (ES) cells are a uniquely self-renewing, pluripotent population of cells that must be differentiated before being useful for cell therapy. Since most studies utilize subcutaneous implantation to test the in vivo functionality of ES cell-derived cells, the objective of the current study was to develop an appropriate and clinically relevant in vivo implantation system in which the behavior and tumorigenicity of ES cell-derived cells could be effectively tested in a tissue-specific (orthotopic) site. Male ES cells were differentiated either into osteoblasts or chondrocytes using protocols that were previously developed and published by our laboratory. The differentiated cells were implanted into a burr-hole fracture created in the proximal tibiae of immunocompetent female mice, strain matched to the ES cell line. The ability of the differentiated ES cell-derived cells (bearing the Y chromosome) to incorporate into the newly formed bone was assessed by micro-computed tomography imaging and histochemistry. ES cells differentiated with either osteogenic or chondrogenic medium supplementation formed a soft tissue mass that disrupted the normal bone architecture by 4 weeks after implantation in some mice. In contrast, mice receiving osteoblastic cells that were differentiated in a three-dimensional type 1 collagen gel showed evidence of new bone formation at the defect site without evidence of tumor formation for up to 8 weeks after implantation. In this injury model, type 1 collagen is more effective than medium supplementation at driving more complete differentiation of ES cells, as evidenced by reducing their tumorigenicity. Overall, the current study emphasizes the importance of using an appropriate orthotopic implantation system to effectively test the behavior and tumorigenicity of the cells in vivo.

Key words: Embryonic stem (ES) cells; Differentiation; Collagen; Orthotopic model; Implantation; Skeletal tissue

Received April 17, 2011; final acceptance June 26, 2011. Online prepub date: October 31, 2012.
Address correspondence to Jaymi Taiani, Biomedical Engineering, Faculty of Medicine, University of Calgary, McCaig Institute of Bone and Joint Health, 3330 Hospital Drive NW, Calgary, AB, Canada, T2N 4N1. Tel: +1-403-220-4549; Fax: +1-403-270-3679; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1463-1476, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657341
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Enhanced Oxygen Supply Improves Islet Viability in a New Bioartificial Pancreas

Uriel Barkai,* Gordon C. Weir,† Clark K. Colton,‡ Barbara Ludwig,§ Stefan R. Bornstein,§ Mathias D. Brendel,§ Tova Neufeld,* Chezi Bremer,* Assaf Leon,* Yoav Evron,* Karina Yavriyants,* Dimitri Azarov,* Baruch Zimermann,* Shiri Maimon,* Noa Shabtay,* Maria Balyura,* Tania Rozenshtein,* Pnina Vardi,¶ Konstantin Bloch,# Paul de Vos,** and Avi Rotem*

*Beta-O2 Technologies, Kiryat Arie, Petach Tikva, Israel
†Section of Islet Transplantation and Cell Biology, Joslin Diabetes Center, Research Division, Boston, MA, USA
‡Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
§University Hospital Carl Gustav Carus, Department of Medicine III, Dresden, Germany
¶Department of Diabetes, Lin Medical Center, Clalit Health Services, Haifa, Israel
#Diabetes and Obesity Research Laboratory, Felsenstein Medical Research Center, Sackler Faculty of Medicine, Tel-Aviv University, Beilinson Campus, Petah Tikva, Israel
**Department of Pathology and Laboratory Medicine, Section of Immunoendocrinology, University Medical Center Groningen, Groningen, The Netherlands

The current epidemic of diabetes with its overwhelming burden on our healthcare system requires better therapeutic strategies. Here we present a promising novel approach for a curative strategy that may be accessible for all insulin-dependent diabetes patients. We designed a subcutaneous implantable bioartificial pancreas (BAP)—the “β-Air”—that is able to overcome critical challenges in current clinical islet transplantation protocols: adequate oxygen supply to the graft and protection of donor islets against the host immune system. The system consists of islets of Langerhans immobilized in an alginate hydrogel, a gas chamber, a gas permeable membrane, an external membrane, and a mechanical support. The minimally invasive implantable device, refueled with oxygen via subdermally implanted access ports, completely normalized diabetic indicators of glycemic control (blood glucose intravenous glucose tolerance test and HbA1c) in streptozotocin-induced diabetic rats for periods up to 6 months. The functionality of the device was dependent on oxygen supply to the device as the grafts failed when oxygen supply was ceased. In addition, we showed that the device is immunoprotective as it allowed for survival of not only isografts but also of allografts. Histological examination of the explanted devices demonstrated morphologically and functionally intact islets; the surrounding tissue was without signs of inflammation and showed visual evidence of vasculature at the site of implantation. Further increase in islets loading density will justify the translation of the system to clinical trials, opening up the potential for a novel approach in diabetes therapy.

Key words: Diabetes; Bioartificial pancreas (BAP); Islets; Implantation; Immune barrier; Oxygen

Received January 18, 2012; final acceptance July 18, 2012. Online prepub date: October 3, 2012.
Address correspondence to Uriel Barkai, Ph.D., Beta-O2 Technologies, 6 Efal st, POB 3226, Kiryat Arie, Petach Tikva 49511, Israel. Tel: +972 3 918 0700; Fax: +972 3 918 0701; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1477-1483, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657639
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Quality of Isolated Pig Islets Is Improved Using Perfluorohexyloctane for Pancreas Storage in a Split Lobe Model

H. Brandhorst,*† M. Iken,‡ W. E. Scott III,§¶ K. K. Papas,§¶ B. Theisinger,# P. R. Johnson,† O. Korsgren,* and D. Brandhorst*†

*Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
†Nuffield Department of Surgical Sciences, University of Oxford, Oxford, UK
‡Department of Gastroenterology, Hepatology and Endocrinology, University Hospital Hannover, Hannover, Germany
§Schulze Diabetes Institute for Immunology and Transplantation, Department of Surgery, University of Minnesota, Minneapolis, MN, USA
¶Institute for Cellular Transplantation, University of Arizona, Tuscon, AZ, USA
#Novaliq GmbH, Heidelberg, Germany

Pancreas transportation between donor center and islet production facility is frequently associated with prolonged ischemia impairing islet isolation and transplantation outcomes. It is foreseeable that shipment of pig pancreases from distant centralized biosecure breeding facilities to institutes that have a long-term experience in porcine islet isolation is essentially required in future clinical islet xenotransplantation. Previously, we demonstrated that perfluorohexyloctan (F6H8) is significantly more efficient to protect rat and human pancreata from ischemically induced damage compared to perfluorodecalin (PFD). To evaluate the effect of F6H8 on long-term stored pig pancreases in a prospective study, we utilized the split lobe model to minimize donor variability. Retrieved pancreases were dissected into the connecting and splenic lobe, intraductally flushed with UW solution and immersed alternately in either preoxygenated F6H8 or PFD for 8–10 h. Prior to pancreas digestion, the intrapancreatic pO2 and the ratio of ATP-to-inorganic phosphate was compared utilizing 31P-NMR spectroscopy. Isolated islets were cultured for 2–3 days at 37°C and subjected to quality assessment. Pancreatic lobes stored in preoxygenated F6H8 had a significantly higher intrapancreatic pO2 compared to pancreata in oxygen-precharged PFD (10.11 ± 3.87 vs. 1.64 ± 1.13 mmHg, p < 0.05). This correlated with a higher ATP-to-inorganic phosphate ratio (0.30 ± 0.04 vs. 0.14 ± 0.01). No effect was observed concerning yield and purity of freshly isolated islets. Nevertheless, a significantly improved glucose-stimulated insulin response, increased viability and postculture survival (57.2 ± 5.7 vs. 39.3 ± 6.4%, p < 0.01) was measured in islets isolated from F6H8-preserved pancreata. The present data suggest that F6H8 does not increase islet yield but improves quality of pig islets isolated after prolonged cold ischemia.

Key words: Cold storage; Islet isolation; Organ preservation; Oxygenation; Pig pancreas

Received April 3, 2012; final acceptance July 25, 2012. Online prepub date: October 4, 2012.
Address correspondence to Daniel Brandhorst, Nuffield Department of Surgical Sciences, Oxford Center for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Drive, Oxford OX3 7LE, UK. Tel: +44-1865-8-57252; Fax: +44-1865-8-57299; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1485-1499, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653192
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Enhancing the Adhesion of Hematopoietic Precursor Cell Integrins With Hydrogen Peroxide Increases Recruitment Within Murine Gut

Dean P. J. Kavanagh,* Adrian I. Yemm,* J. Steven Alexander,† Jon Frampton,* and Neena Kalia*

*Centre for Cardiovascular Sciences, Institute of Biomedical Research, The Medical School, University of Birmingham, Edgbaston, Birmingham, UK
†Department of Molecular and Cellular Physiology, LSU Health Sciences Centre, Shreveport, LA, USA

Hematopoietic stem cells (HSCs) migrate to injury sites and aid in tissue repair. However, clinical success is poor and is partially due to limited HSC recruitment. We hypothesized that HSC pretreatment with H2O2 would enhance their recruitment to injured gut. As HSCs are rare cells, the number of primary cells obtained from donors is often inadequate for functional experiments. To circumvent this, in this study we utilized a functionally relevant cell line, HPC-7. Anesthetized mice were subjected to intestinal ischemia-reperfusion (IR) injury, and HPC-7 recruitment was examined intravitally. Adhesion to endothelial cells (ECs), injured gut sections, and ICAM-1/VCAM-1 protein were also quantitated in vitro. H2O2 pretreatment significantly enhanced HPC-7 recruitment to injured gut in vivo. A concomitant reduction in pulmonary adhesion was also observed. Enhanced adhesion was also observed in all in vitro models. Increased clustering of a4 and b2 integrins, F-actin polymerization, and filopodia formation were observed in pretreated HPC-7s. Importantly, H2O2 did not reduce HPC-7 viability or proliferative ability. HPC-7 recruitment to injured gut can be modulated by H2O2 pretreatment. This may be through increasing the affinity or avidity of surface integrins that mediate HPC-7 homing to injured sites or through stimulating the migratory apparatus. Strategies that enhance hematopoietic stem/progenitor cell recruitment may ultimately affect their therapeutic efficacy.

Key words: Adhesion molecules; Cell adhesion; Small intestine; Ischemia-reperfusion (IR) injury; Hydrogen peroxide (H2O2)

Received October 25, 2011; final acceptance May 23, 2012. Online prepub date: August 10, 2012.
Address correspondence to Dr. Neena Kalia, Centre for Cardiovascular Sciences, Institute of Biomedical Research, The Medical School, University of Birmingham, Birmingham B15 2TT, UK. Tel.: +44 (0) 121 414 8818; Fax: +44 (0) 121 415 8817; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1501-1506, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657611
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Functional Stability (at +4°C) of Hematopoietic Stem and Progenitor Cells Amplified Ex Vivo From Cord Blood CD34+ Cells

Pascale Duchez,*† Jean Chevaleyre,*† Philippe Brunet de la Grange,*† Marija Vlaski,*† Jean-Michel Boiron,*† and Zoran Ivanovic*†

*Etablissement Français du Sang, Aquitaine-Limousin, Bordeaux, France
†CNRS, Univ. Bordeaux Segalen, CIRID UMR 5164, Bordeaux, France Université Bordeaux Segalen, Bordeaux, France

Our previously published ex vivo expansion procedure starting from cord blood CD34+ cells enables a massive expansion of total and CD34+ cells and committed progenitors without negative impact on stem cells exhibiting both short- and long-term repopulating capacity. It was upgraded to clinical scale [Macopharma HP01® medium in presence of SCF, FLT3-L (100 ng/ml each), G-SCF (10 ng/ml), and TPO (20 ng/ml)] and is in use for an ongoing clinical trial (adult allogeneic context), yielding encouraging results. In order to test the possibility to use the expanded cells in distant transplantation centers, we studied the functional stability at +4°C (usual temperature of transportation) of hematopoietic progenitors and stem cells 48 h after expansion. If the cells were washed and resuspended in 4% albumin solution (actual procedure for immediate injection), only one half of total nucleated and CD34+ cells and 30% of committed progenitors survived after 24 h. This condition has also an evident negative impact on stem cells in expansion product as demonstrated on the basis of reconstitution of NSG mice bone marrow by human CD45, CD33, CD19+ cells as well as by human committed progenitors (CFU). Surprisingly, if the cells were stored 48 h at +4°C in culture medium, very good survival of total and CD34+ cells (90 to 100%) and colony forming unit cells (CFCs; around 70%) was obtained, as well as the maintenance of stem cells (the same in vivo assay with NSG mice). These data point to the possibility of the maintenance of the full functional capacity of expanded grafts for 2 days, the time allowing for its transportation to any transplantation center worldwide.

Key words: Cord blood; Ex vivo expansion; Committed progenitors; Stem cells; Liquid storage

Received March 29, 2012; final acceptance August 1, 2012. Online prepub date: October 4, 2012.
Address correspondence to Zoran Ivanovic, M.D., Ph.D., Etablissement Français du Sang Aquitaine-Limousin, Place Amélie Raba Léon, BP24, 3035 Bordeaux Cedex, France. Tel: +33556907550; Fax: +33556907551; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it