Cell Transplantation 20(6) Abstracts

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Cell Transplantation, Vol. 20, pp. 789–795, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539074
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Amniotic Fluid as a Rich Source of Mesenchymal Stromal Cells for Transplantation Therapy

Ivana Antonucci,* Liborio Stuppia,* Yuji Kaneko,† Seongjin Yu,† Naoki Tajiri,† Eunkyung C. Bae,† Sonia H. Chheda,† Nathan L. Weinbren,† and Cesar V. Borlongan†

*Biomedical Science, Chieti University and Stem TeCh Group, Aging Research Center (CESI), Chieti, Italy
†Department of Neurosurgery and Brain Repair, University of South Florida College of Medicine, Tampa, FL, USA

Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, thus being considered as a powerful tool for cellular therapy of different human diseases. In the last 4 years, amniotic fluid-derived stem (AFS) cells have been shown to express embryonic and adult stem cell markers. These cells can be considered an intermediate stage between embryonic stem cells and adult stem cells. AFS cells can give rise to adipogenic, osteogenic, myogenic, endothelial, neurogenic, and hepatic lineages, inclusive of all embryonic germ layers. AFS cells have a high renewal capacity and can be expanded for over 250 doublings without any detectable loss of chromosomal telomere length. Taken together, all these data provide evidence that amniotic fluid represents a new and very promising source of stem cells for research, as well as clinical applications. Certainly stem cells from amniotic fluid will be useful both for a customized cell supply for newly born children and for banking cells to be used for therapeutic cell transplantation in immunogically matched recipients. Further investigations are also warranted to fully explore the amniotic cells’ potential for adult human disorders.

Key words: Stem cells; Placenta; Transplantation; Human disorders

Address correspondence to Cesar V. Borlongan, Ph.D., Professor and Vice-chairman for Research, Department of Neurosurgery and Brain Repair, University of South Florida, Tampa, FL 33612, USA. Tel: 1-813-974-3154; Fax: 1-813-974-3078l; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 20, pp. 797–811, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X543376
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Platelet Lysate Consisting of a Natural Repair Proteome Supports Human Mesenchymal Stem Cell Proliferation and Chromosomal Stability

Ruben Crespo-Diaz,* Atta Behfar,* Greg W. Butler,† Douglas J. Padley,† Michael G. Sarr,‡ Jozef Bartunek,§ Allan B. Dietz,† and Andre Terzic*

*Marriott Heart Disease Research Program, Division of Cardiovascular Diseases, Departments of Medicine and Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN, USA
†Human Cellular Therapy Laboratory, Division of Transfusion Medicine, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA
‡Department of General Surgery, Mayo Clinic, Rochester, MN, USA
§Cardiovascular Center, Aalst, Belgium

With favorable regenerative and immunotolerant profiles, patient-derived human mesenchymal stem cells (hMSCs) are increasingly considered in cell therapy. Derived from bone marrow (BM) and standardized with culture in fetal bovine serum (FBS), translation of hMSC-based approaches is impeded by protracted expansion times, risk of xenogenic response, and exposure to zoonoses. Here, human platelet lysate adherent to good manufacturing practices (GMP-hPL) provided a nonzoonotic adjuvant that enhanced the capacity of BM-hMSC to proliferate. The nurturing benefit of GMP-hPL was generalized to hMSC from adipose tissue evaluated as an alternative to bone marrow. Long-term culture in GMP-hPL maintained the multipotency of hMSC, while protecting against clonal chromosomal instability detected in the FBS milieu. Proteomic dissection identified TGF-β, VEGF, PDGF, FGF, and EGF as highly ranked effectors of hPL activity, revealing a paradigm of healing that underlies platelet lysate adjuvancy. Thus, GMP-adherent human platelet lysate accelerates hMSC proliferation with no chromosomal aberrancy, through an innate repair paradigm.

Key words: Human mesenchymal stem cells (hMSCs); Human platelet lysate (hPL); Chromosomal stability

Address correspondence to Atta Behfar, Mayo Clinic, Stabile 5, 200 1st St. SW, Rochester, MN 55905, USA. Tel: +1 507-284-9012; Fax: +1 507-266-9936; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 813–825, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539100
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Glucose Reduction Prevents Replicative Senescence and Increases Mitochondrial Respiration in Human Mesenchymal Stem Cells

Ting Lo,* Jennifer H. Ho,†‡ Muh-Hwa Yang,*§ and Oscar K. Lee*¶#

*Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
†Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan
‡Department of Ophthalmology, Wan Fang Hospital, Taipei, Taiwan
§Division of Hematology-Oncology, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan
¶Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan
#Stem Cell Research Center, National Yang-Ming University, Taipei, Taiwan

The unique self-renewal and multilineage differentiation potential of mesenchymal stem cells (MSCs) make them a promising candidate for cell therapy applications. However, during in vitro expansion of MSCs, replicative senescence may occur and will compromise the quality of the expanded cells. Because calorie restriction has been shown to effectively extend the life span of various organisms, the purpose of this study is to investigate the effect of glucose reduction on MSCs and the coordinated changes in energy utilization. It was found that the frequency of cycling cells was significantly increased, while senescence markers such as β-galactosidase activities and p16INK4a expression level were markedly reduced in MSCs under low-glucose culture condition. Quantitative real-time PCR analysis demonstrated the preserved trilineage differentiation potentials of MSCs after low-glucose treatment. Interestingly, the ability of osteogenic lineage commitment was improved, while the ability of adipogenic lineage commitment was delayed in MSCs after glucose reduction. In addition, we observed decreased lactate production, increased electron transport chain complexes expression, and increased oxygen consumption in MSCs after glucose reduction treatment. Increased antioxidant defensive responses were evidenced by increased antioxidant enzymes expression and decreased superoxide production after glucose reduction. Taken together, our findings suggest that MSCs utilize energy more efficiently under restricted glucose treatment and exhibit greater self-renewal and antisenescence abilities, while their differentiation potentials remain unaffected.

Key words: Glucose reduction; Mesenchymal stem cells (MSCs); Replicative senescence; Mitochondria

Address correspondence to Oscar K. Lee, M.D., Ph.D., Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road,Taipei, Taiwan. Tel: +886-2-28757391; Fax: +886-2-28757409; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 827–835, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X540630
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Neuroprotective Effect of Human Mesenchymal Stem Cells in an Animal Model of Double Toxin-Induced Multiple System Atrophy Parkinsonism

Hyun-Jung Park,*† Giyoon Bang,‡ Bo Ra Lee,‡ Hyun Ok Kim,§ and Phil Hyu Lee†‡

*Neuroscience Graduate Program, Ajou University School of Medicine, Suwon, South Korea
†Severance Biomedical Science Institute, Yonsei University, Seoul, South Korea
‡Department of Neurology, Yonsei University College of Medicine, Seoul, South Korea
§Department of Laboratory Medicine, Yonsei Cell Therapy Center, Yonsei University College of Medicine, Seoul, South Korea

Multiple system atrophy (MSA) is an adult-onset sporadic neurodegenerative disorder of unknown etiology featuring parkinsonism, ataxia, and autonomic failure in any combination. Because disease progression in MSA is rapid and no drug treatment consistently benefits MSA patients in the long term, neuroprotective or regenerative strategies may be invaluable in the management of MSA patients. In this study, we investigated whether human mesenchymal stem cells (hMSCs) had a protective effect on MSA using an animal model of double-toxin-induced MSA parkinsonism (MSA-P). MSA-P was established with coinjections of MPTP and 3-NP; hMSCs were injected into the tail vein 1 day after the last toxin injection. Three groups of mice were compared (i.e., control, MPTP + 3-NP, and MPTP + 3-NP with hMSC treatment) through histopathological, behavioral, and Western blot analyses. In the substantia nigra (SN) and the striatum, 2.0% and 3.8% of total injected hMSCs were observed, respectively. Compared with double-toxin-treated mice, hMSC treatment in double-toxin-treated mice significantly increased survival of TH- and NeuN-immunoreactive cells in the SN and the striatum, with coincident improvement in motor behavior. Additionally, hMSC treatment significantly decreased double-toxin-induced microglial and astroglial activation in the SN and striatum. Western blot analysis showed that hMSC administration in double-toxin-treated mice increased the expression of p-Akt and Bcl-2 and decreased Bax and cytochrome c expression. This study demonstrates that hMSC treatment protected against loss of neurons in the SN and the striatum induced by double toxin exposure, which may be mediated by modulation of inflammatory and cell survival and death signalingpathway as the hMSCs migrated from the peripheral circulation into the SN and striatum.

Key words: Multiple system atrophy (MSA); Mesenchymal stem cells (MSCs); Neuroprotection

Address correspondence to Phil Hyu Lee, M.D., Ph.D., Department of Neurology, Yonsei University College of Medicine, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-752, South Korea. Tel: 82-2-2228-1608; Fax: 82-2-393-0705; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 837–849, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539083
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Cotransplantation of Mouse Neural Stem Cells (mNSCs) With Adipose Tissue-Derived Mesenchymal Stem Cells Improves mNSC Survival in a Rat Spinal Cord Injury Model

Jin Soo Oh,*† Keung Nyun Kim,* Sung Su An,*† William A. Pennant,* Hyo Jin Kim,*† So-Jung Gwak,*† Do Heum Yoon,*† Mi Hyun Lim,‡ Byung Hyune Choi,‡ and Yoon Ha*

*Department of Neurosurgery, Spine & Spinal Cord Institute, College of Medicine, Yonsei University, Seoul, Korea
†Brain Korea 21 Project for Medical Science, College of Medicine, Yonsei University, Seoul, Korea
‡Division of Biomedical and Bioengineering Sciences, Inha University College of Medicine, Incheon, Korea

The low survival rate of graft stem cells after transplantation into recipient tissue is a major obstacle for successful stem cell therapy. After transplantation into the site of spinal cord injury, the stem cells face not only hypoxia due to low oxygen conditions, but also a lack of nutrients caused by damaged tissues and poor vascular supply. To improve the survival of therapeutic stem cells after grafting into the injured spinal cord, we examined the effects of cotransplanting mouse neural stem cells (mNSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs) on mNSC viability. The viability of mNSCs in coculture with ATMSCs was significantly increased compared to mNSCs alone in an in vitro injury model using serum deprivation (SD), hydrogen peroxide (H2O2), and combined (SD + H2O2) injury mimicking the ischemic environment of the injured spinal cord. We demonstrated that AT-MSCs inhibited the apoptosis of mNSCs in SD, H2O2, and combined injury models. Consistent with these in vitro results, mNSCs transplanted into rat spinal cords with AT-MSCs showed better survival rates than mNSCs transplanted alone. These findings suggest that cotransplantation of mNSCs with AT-MSCs may be a more effective transplantation protocol to improve the survival of cells transplanted into the injured spinal cord.

Key words: Mouse neural stem cells (mNSCs); Adipose tissue-derived mesenchymal stem cells (AT-MSCs); Spinal cord injury

Address correspondence to Yoon Ha, M.D., Ph.D., Department of Neurosurgery, Spine and Spinal Cord Institute, Yonsei University College of Medicine, 120-752, 134 Shinchon-dong, Seodaemoon-gu, Seoul, Korea. Tel: +82-2-2228-2165; Fax: +82-2-393-9970; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 851–869, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X543411
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Clinical Potential of Intravenous Neural Stem Cell Delivery for Treatment of Neuroinflammatory Disease in Mice?

Kristien P. Reekmans,*†‡ Jelle Praet,*†§ Nathalie De Vocht,*†§ Bart R. Tambuyzer,*† Irene Bergwerf,*† Jasmijn Daans,*† Veerle Baekelandt,¶# Greetje Vanhoutte,§ Herman Goossens,† Philippe G. Jorens,‡** Dirk K. Ysebaert,‡†† Shyama Chatterjee,‡‡ Patrick Pauwels,‡‡ Eric Van Marck,‡‡ Zwi N. Berneman,*†‡ Annemie Van der Linden,§ and Peter Ponsaerts*

*Laboratory of Experimental Hematology, University of Antwerp, Antwerp, Belgium
†Vaccine and Infectious Disease Institute, University of Antwerp, Antwerp, Belgium
‡Centre for Cell Therapy and Regenerative Medicine, Antwerp University Hospital, Edegem, Belgium
§BioImaging Laboratory, University of Antwerp, Antwerp, Belgium
¶Laboratory for Neurobiology & Gene Therapy, Katholieke Universiteit Leuven, Leuven, Belgium
#Molecular Small Animal Imaging Centre (MoSAIC), Katholieke Universiteit Leuven, Leuven, Belgium
**Clinical Pharmacotherapy, University of Antwerp, Antwerp, Belgium
††Laboratory of Experimental Surgery, University of Antwerp, Antwerp, Belgium
‡‡Laboratory of Pathology, University of Antwerp, Antwerp, Belgium

While neural stem cells (NSCs) are widely expected to become a therapeutic agent for treatment of severe injuries to the central nervous system (CNS), currently there are only few detailed preclinical studies linking cell fate with experimental outcome. In this study, we aimed to validate whether IV administration of allogeneic NSC can improve experimental autoimmune encephalomyelitis (EAE), a well-established animal model for human multiple sclerosis (MS). For this, we cultured adherently growing luciferase-expressing NSCs (NSC-Luc), which displayed a uniform morphology and expression profile of membrane and intracellular markers, and which displayed an in vitro differentiation potential into neurons and astrocytes. Following labeling with green fluorescent micron-sized iron oxide particles (f-MPIO-labeled NSC-Luc) or lentiviral transduction with the enhanced green fluorescent protein (eGFP) reporter gene (NSC-Luc/eGFP), cell implantation experiments demonstrated the intrinsic survival capacity of adherently cultured NSC in the CNS of syngeneic mice, as analyzed by real-time bioluminescence imaging (BLI), magnetic resonance imaging (MRI), and histological analysis. Next, EAE was induced in C57BL/6 mice followed by IV administration of NSC-Luc/eGFP at day 7 postinduction with or without daily immunosuppressive therapy (cyclosporine A, CsA). During a follow-up period of 20 days, the observed clinical benefit could be attributed solely to CsA treatment. In addition, histological analysis demonstrated the absence of NSC-Luc/eGFP at sites of neuroinflammation. In order to investigate the absence of therapeutic potential, BLI biodistribution analysis of IV-administered NSC-Luc/eGFP revealed cell retention in lung capillaries as soon as 1-min postinjection, resulting in massive inflammation and apoptosis in lung tissue. In summary, we conclude that IV administration of NSCs currently has limited or no therapeutic potential for neuroinflammatory disease in mice, and presumably also for human MS. However, given the fact that grafted NSCs have an intrinsic survival capacity in the CNS, their therapeutic exploitation should be further investigated, and—in contrast to several other reports—will most likely be highly complex.

Key words: Neural stem cells (NSCs); Transplantation; Bioluminescence imaging; Experimental autoimmune encephalomyelitis; Cell migration

Address correspondence to Dr. Peter Ponsaerts, Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Campus Drie Eiken (CDE-T1.17), Universiteitsplein 1, 2610 Antwerp (Wilrijk), Belgium. Tel: 0032-3-2652428; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 871–881, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539065
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Light Deprivation Induces Depression-Like Behavior and Suppresses Neurogenesis in Diurnal Mongolian Gerbil (Meriones unguiculatus)

Benson Wui-Man Lau,* Chaoran Ren,† Jian Yang,* Sylvia W. L. Yan,* Raymond Chuen-Chung Chang,*‡ Mingliang Pu,† and Kwok-Fai So*§

*Department of Anatomy, The State Key Laboratory of Brain and Cognitive Sciences, Research Centre of Heart, Brain, Hormone and Healthy Aging, The University of Hong Kong, Pokfulam, Hong Kong SAR, PR China
†Department of Anatomy and Embryology, School of Basic Medical Sciences, Key Laboratory on Machine Perception, Peking University, Beijing, PR China
‡Laboratory of Neurodegenerative Diseases, Department of Anatomy, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, PR China
§Joint laboratory for Brain function and Health (BFAH), Jinan University and The University of Hong Kong, GuangZhou, PR China

Recent evidence suggests that adult neurogenesis contributes to the pathophysiology of different psychiatric disorders, including depressive disorder, anxiety disorder, and schizophrenia. Seasonal affective disorder (SAD) is a specific form of recurrent depressive disorder that can be induced by shortened light period. It is unclear yet whether neurogenesis is affected in SAD or under altered light/dark cycle. The present study aims at examining whether neurogenesis and dendritic growth of immature neurons are affected in Mongolian gerbils, a mainly diurnal rodent, under light deprivation. Animals were divided into two groups: the control (kept in 12 h light:12 h dark) and the light-deprived groups (kept in 24 h dark). Depression-like behaviors and neurogenesis were assessed after 2 weeks. Compared with the control group, light-deprived gerbils showed increased immobile time in the tail suspension test and forced swimming test, which indicates induction of depression-like behavior. Cell proliferation in both the hippocampal and subventricular zone were significantly decreased in the light-deprived group, which also showed a decreased neuronal differentiation. Dendritic maturation of immature neurons was suppressed by light deprivation, which is revealed by doublecortin staining and Sholl analysis. The results revealed that the light/dark cycle exerts impacts on neurogenesis and maturation of new neurons. Additionally, the current experiment may offer a model for exploring the relationship among daylight exposure, circadian cycles, depressive behavior, and the underlying mechanisms.

Key words: Seasonal affective disorder (SAD); Neurogenesis; Light deprivation; Hippocampus; Subventricular zone; Mongolian gerbil; Diurnal

Address correspondence to Professor Kwok-Fai So, Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Pokfulam, Hong Kong SAR, PR China. Tel: (852) 2819-9216; Fax: (852) 2817-0857; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Professor Mingliang Pu, Department of Anatomy and Embryology, School of Basic Medical Sciences, Peking University, Beijing, PR China. Tel: 86-10- 82802972; Fax: 010-8280-2877; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 883–891, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539092
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Tumorigenic Development of Induced Pluripotent Stem Cells in Ischemic Mouse Brain

Toru Yamashita, Hiromi Kawai, Fengfeng Tian, Yasuyuki Ohta, and Koji Abe

Department of Neurology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

Induced pluripotent stem (iPS) cells may provide cures for various neurological diseases. However, undifferentiated iPS cells have high tumorigenicity, and evaluation of the cells fates, especially in pathologic condition model, is needed. In this study, we demonstrated the effect of ischemic condition to undifferentiated iPS cells fates in a mouse model of transient middle cerebral artery occlusion (MCAO). Undifferentiated iPS cells were characterized with immunofluorescent staining. The iPS cells (5 × 105) were injected into ipsilateral striatum and cortex after 24 h of MCAO. Histological analysis was performed from 3 to 28 days after cell transplantation. iPS cells in ischemic brain formed teratoma with higher probability (p < 0.05) and larger volume (p < 0.01) compared with those in intact brain. Among the four transcriptional factors to produce iPS cells, c-Myc, Oct3/4, and Sox2 strongly expressed in iPS-derived tumors in ischemic brain (p < 0.01). Additionally, expression of matrix metalloproteinase-9 (MMP-9) and phosphorylated vascular endothelial growth factor receptor2 (phospho-VEGFR2) were significantly increased in iPS-derived tumors in the ischemic brain (p < 0.05). These results suggest that the transcriptional factors might increase expression of MMP-9 and activate VEGFR2, promoting teratoma formation in the ischemic brain. We strongly propose that the safety of iPS cells should be evaluated not only in normal condition, but also in a pathologic, disease model.

Key words: Induced pluripotent stem (iPS) cells; Cerebral ischemia; Tumorigenesis; Cell transplantation

Address correspondence to Koji Abe, Department of Neurology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. Tel: +81-86-235-7365; Fax: +81-86-235-7368; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 893–907, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X550206
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Collagen IV Significantly Enhances Migration and Transplantation of Embryonic Stem Cells: Involvement of α2β1 Integrin-Mediated Actin Remodeling

Hsin-Yang Li,*†‡ Chen-Yi Liao,‡ Kun-Hsiung Lee,§ Hung-Chi Chang,* Yi-Jen Chen,*† Kuan-Chong Chao,*† Sheng-Ping Chang,*† Hsin-Yi Cheng,*† Chia-Ming Chang,*† Yuh-Lih Chang,¶# Shih-Chieh Hung,# Yen-Jen Sung,‡ and Shih-Hwa Chiou¶#

*Department of Obstetrics and Gynecology, Taipei Veterans General Hospital, Taipei, Taiwan
†Division of Obstetrics and Gynecology, Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan
‡Institute of Anatomy and Cell Biology, School of Medicine, National Yang-Ming University, Taipei, Taiwan
§Division of Biotechnology, Animal Technology Institute Taiwan, Chunan, Miaoli, Taiwan
¶Institute of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan
#Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan

Embryonic stem (ES) cell transplantation represents a potential means for the treatment of degenerative diseases and injuries. As appropriate distribution of transplanted ES cells in the host tissue is critical for successful transplantation, the exploration of efficient strategies to enhance ES cell migration is warranted. In this study we investigated ES cell migration under the influence of various extracellular matrix (ECM) proteins, which have been shown to stimulate cell migration in various cell models with unclear effects on ES cells. Using two mouse ES (mES) cell lines, ESC 26GJ9012-8-2 and ES-D3 GL, to generate embryoid bodies (EBs), we examined the migration of differentiating cells from EBs that were delivered onto culture surfaces coated with or without collagen I, collagen IV, Matrigel, fibronectin, and laminin. Among these ECM proteins, collagen IV exhibited maximal migration enhancing effect. mES cells expressed α2 and β1 integrin subunits and the migration enhancing effect of collagen IV was prevented by RGD peptides as well as antibodies against α2 and β1 integrins, indicating that the enhancing effect of collagen IV on cell migration was mediated by α2β1 integrin. Furthermore, staining of actin cytoskeleton that links to integrins revealed well-developed stress fibers and long filopodia in mES cells cultured on collagen IV, and the actindisrupting cytochalasin D abolished the collagen IV-enhanced cell migration. In addition, pretreatment of undifferentiated or differentiated mES cells with collagen IV resulted in improved engraftment and growth after transplantation into the subcutaneous tissue of nude mice. Finally, collagen IV pretreatment of osteogenically differentiated mES cells increased osteogenic differentiation-like tissue and decreased undifferentiation- like tissue in the grafts grown after transplantation. Our results demonstrated that collagen IV significantly enhanced the migration of differentiating ES cells through α2β1 integrin-mediated actin remodeling and could promote ES cell transplantation efficiency, which may be imperative to stem cell therapy.

Key words: Extracellular matrix (ECM); Collagen IV; Embryonic stem (ES) cells; Migration; Transplantation; Integrins

Address correspondence to Yen-Jen Sung, M.D., Ph.D., Institute of Anatomy and Cell Biology, School of Medicine, National Yang-Ming University, 155 Li-Nong Street Section 2, Taipei 112, Taiwan. Tel: +886-2-28267000; Fax: +886-2-28212884; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Shih-Hwa Chiou, M.D., Ph.D., Institute of Pharmacology, School of Medicine, National Yang-Ming University, 155 Li-Nong Street Section 2, Taipei 112, Taiwan. Tel: +886-2-28757394; Fax: +886-2-28734101; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 909–923, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X543385
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Amnion Epithelial Cells Prevent Bleomycin-Induced Lung Injury and Preserve Lung Function

Sean Murphy,* Rebecca Lim,* Hayley Dickinson,* Rutu Acharya,* Sharina Rosli,* Graham Jenkin,* and Euan Wallace*†

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

Human amnion epithelial cells (hAECs) have attracted recent attention as a promising source of cells for regenerative therapies, with reports that cells derived from human term amnion possess multipotent differentiation ability, low immunogenicity, and anti-inflammatory properties. Specifically, in animal models of lung disease characterized by significant loss of lung tissue secondary to chronic inflammation and fibrosis, the transplantation of hAECs has been shown to reduce both inflammation and subsequent fibrosis. To further explore the mechanisms by which hAECs reduce pulmonary fibrosis and enhance lung regeneration, we utilized a bleomycin-induced model of pulmonary fibrosis and investigated the ability of hAECs to reduce fibrosis and thereby improve pulmonary function. We aimed to determine if hAECs, injected into the peritoneal cavity could migrate to the lung, engraft, and form functional lung epithelium, and whether hAECs could modulate the inflammatory environment in the bleomycin-injured lung. We demonstrated that, compared to bleomycin alone, IP administration of hAECs 24 h after bleomcyin, decreased gene expression of the proinflammatory cytokines TNF-α, TGF-β, IFN-γ, and IL-6 and decreased subsequent pulmonary fibrosis with less pulmonary collagen deposition, reduced levels of α-smooth muscle actin and decreased inflammatory cell infiltrate. We also showed that hAECs are able to prevent a decline in pulmonary function associated with bleomycin-induced lung damage. We were unable to detect any significant engraftment of hAECs in injured, or uninjured, lung after administration. The findings from this study support the further investigation of hAECs as a potential cell therapy for inflammatory and fibrogenic diseases.

Key words: Amnion epithelial cells (AECs); Lung disease; Inflammation; Fibrosis; Lung function

Address correspondence to Professor Euan M. Wallace, Director, The Ritchie Centre, Department of Obstetrics and Gynaecology, Level 5, Monash Medical Centre, 246 Clayton Road, Clayton, Vic, 3168, Australia. Tel: +613 9594 5145; Fax: +613 9594 5003; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 925–940, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539119
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Differences in Cartilage-Forming Capacity of Expanded Human Chondrocytes From Ear and Nose and Their Gene Expression Profiles

Catharine A. Hellingman,* Eugène T. P. Verwiel,† Inez Slagt,*‡ Wendy Koevoet,* René M. L. Poublon,* Gilbert J. Nolst-Trenite´,§ Robert J. Baatenburg de Jong,* Holger Jahr,‡ and Gerjo J. V. M. van Osch*‡

*Department of Otorhinolaryngology, Head and Neck surgery, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands
†Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
‡Department of Orthopaedics, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands
§Department of Otorhinolaryngology, Academic Medical Center (AMC), Amsterdam, The Netherlands

The aim of this study was to evaluate the potential of culture-expanded human auricular and nasoseptal chondrocytes as cell source for regeneration of stable cartilage and to analyze the differences in gene expression profile of expanded chondrocytes from these specific locations. Auricular chondrocytes in monolayer proliferated less and more slowly (two passages took 26.7 ± 2.1 days and were reached in 4.37 ± 0.30 population doublings) than nasoseptal chondrocytes (19.3 ± 2.5 days; 5.45 ± 0.20 population doublings). However, auricular chondrocytes produced larger pellets with more cartilage-like matrix than nasoseptal chondrocytes (2.2 ± 0.71 vs. 1.7 ± 0.13 mm in diameter after 35 days of culture). Although the matrix formed by auricular and nasoseptal chondrocytes contained collagen X, it did not mineralize in an in vitro model or after in vivo subcutaneous implantation. A DNA microarray study on expanded auricular and nasoseptal chondrocytes from the same donors revealed 1,090 differentially expressed genes. No difference was observed in the expression of known markers of chondrogenic capacity (e.g., collagen II, FGFR3, BMP2, and ALK1). The most striking differences were that the auricular chondrocytes had a higher expression of anabolic growth factors BMP5 and IGF1, while matrix-degrading enzymes MMP13 and ADAMTS5 were higher expressed in nasoseptal chondrocytes. This might offer a possible explanation for the observed higher matrix production by auricular chondrocytes. Moreover, chondrocytes isolated from auricular or nasoseptal cartilage had specific gene expression profiles even after expansion. These differently expressed genes were not restricted to known characterization of donor site subtype (e.g., elastic), but were also related to developmental processes.

Key words: Tissue engineering; Chondrogenesis; Chondrocytes; Cartilage; Collagen X; Mineralization

Address correspondence to Gerjo J. V. M. van Osch, Department of Otorhinolaryngology and Department of Orthopaedics, Erasmus MC, University Medical Center Rotterdam, Room Ee1655, PO Box 2040, 3000 CA Rotterdam, The Netherlands. Tel: +31(0)107043661; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 941–954, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X540621
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

In Vivo Induction of Myeloid Suppressor Cells and CD4+Foxp3+ T Regulatory Cells Prolongs Skin Allograft Survival in Mice

D. Adeegbe,* P. Serafini,*† V. Bronte,‡§ A. Zoso,* C. Ricordi,* and L. Inverardi*¶

*Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, FL, USA
†Department of Microbiology and Immunology, Sylvester Comprehensive Cancer Center, Miller School of Medicine, University of Miami, Miami, FL, USA
‡Verona University Hospital and Department of Pathology, Immunology Section, University of Verona, Verona, Italy
§Istituto Oncologico Veneto, Padova, Italy
¶Division of Endocrinology Diabetes and Metabolism, Miller School of Medicine, University of Miami, Miami, FL, USA

Natural CD4+Foxp3+ T regulatory (Treg) cells can promote transplantation acceptance across major histocompatibility complex (MHC) barriers, while myeloid-derived suppressor cells (MDSCs) inhibit effector Tcell responses in tumor-bearing mice. One outstanding issue is whether combining the potent suppressive function of MDSCs with that of Treg cells might synergistically favor graft tolerance. In the present study, we evaluated the therapeutic potential of MDSCs and natural Treg cells in promoting allograft tolerance in mice by utilizing immunomodulatory agents to expand these cells in vivo. Upon administration of recombinant human granulocyte-colony stimulating factor (G-CSF; Neupogen), or interleukin-2 complex (IL-2C), Gr-1+CD11b+ MDSCs or CD4+Foxp3+ Treg cells were respectively induced at a high frequency in the peripheral lymphoid compartments of treated mice. Interestingly, induced MDSCs exhibited a more potent suppressive function in vitro when compared to MDSCs from naive mice. Furthermore, in vivo coadministration of Neupogen and IL-2C induced MDSCs at percentages that were higher than those seen when either agent was administered alone, suggesting an additive effect of the two drugs. Although treatment with either IL-2C or Neupogen led to a significant delay of MHC class II disparate allogeneic donor skin rejection, the combinatorial treatment was superior to either alone. Importantly, histological assessment of surviving grafts revealed intact morphology and minimal infiltrates at 60 days posttransplant. Collectively, our findings demonstrate that concurrent induction of MDSCs and Tregs is efficacious in downmodulating alloreactive T-cell responses in a synergistic manner and highlight the therapeutic potential of these naturally occurring suppressive leukocytes to promote transplantation tolerance.

Key words: Transplantation; Tolerance; Suppression; Myeloid-derived suppressor cells (MDSCs)

Address correspondence to L. Inverardi, Diabetes Research Institute, Miller School of Medicine, 1450 NW 10th avenue, Miami, FL 33136, USA. Tel: 305-243-5347; Fax: 305-243-4404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 955–962, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539137
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Detecting Rejection After Mouse Islet Transplantation Utilizing Islet Protein-Stimulated ELISPOT

Christian Toso,* Rena Pawlick,† Stéphanie Lacotte,* Ryan Edgar,† Joy Davis,† Michael McCall,† Philippe Morel,* Gilles Mentha,* Thierry Berney,* and A. M. James Shapiro†

*Transplant Unit, Department of Surgery, University of Geneva Hospitals, Geneva, Switzerland
†Section of Hepatobiliary, Pancreatic and Transplant Surgery, University of Alberta, Edmonton, Canada

Improved posttransplant monitoring and on-time detection of rejection could improve islet transplantation outcome. The present study explored the possibility of detecting harmful events after mouse islet transplantation measuring the immune responsiveness against islet extracts. Mouse islet transplantations were performed using various donor/recipient combinations, exploring autoimmune (NOD/SCID to NOD, n = 6) and alloimmune events (C57BL/6 to BALB/c, n = 20), a combination of both (C57BL/6 to NOD, n = 8), the absence of both (BALB/c to BALB/c, n = 21), or naive, nontransplanted control mice (n = 14). The immune reactivity was measured by ELISPOT, looking at the ex vivo release of IFN-γ from splenocytes stimulated by islet donor extracts (sonicated islets). The immune reactivity was not altered in the syngeneic and autoimmune models, demonstrating similar levels as nontransplanted controls (p = 0.46 and p = 0.6). Conversely, the occurrence of an allogeneic rejection alone or in combination to autoimmunity was associated to an increase in the level of immune reactivity (p = 0.023 and p = 0.003 vs. respective controls). The observed increase was transient and lost in the postrejection period or after treatment with CTLA4-Ig. Overall, allogeneic rejection was associated to a transient increase in the reactivity of splenocytes against islet proteins. Such a strategy has the potential to improve islet graft monitoring in human and should be further explored.

Key words: Monitoring; Diabetes; Islets; Transplantation; Cytokine

Address correspondence to Christian Toso, Transplantation Unit, Geneva University Hospitals, rue Gabrielle-Perret-Gentil 4, 1211 Genève 14,Switzerland. Tel: +41 22 372 33 11; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 963–975, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X543367
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Use of a Clinically Approved Iron Oxide MRI Contrast Agent to Label Human Hepatocytes

Juliana Puppi,* Ragai R. Mitry,* Michel Modo,† Anil Dhawan,*‡ Kishor Raja,§ and Robin D. Hughes*

*Institute of Liver Studies, King’s College London School of Medicine at King’s College Hospital, London, UK
†Department of Neuroscience, Institute of Psychiatry, King’s College London, London, UK
‡Paediatric Liver Centre, King’s College Hospital, London, UK
§Department of Clinical Biochemistry, King’s College Hospital, London, UK

Reliable noninvasive methods are needed to monitor cell engraftment and graft survival after hepatocyte transplantation. Superparamagnetic iron oxide nanoparticles (SPIOs) have been shown to accumulate in various types of cells, and are currently the labeling agent of choice for cellular magnetic resonance imaging (MRI). However, for successful clinical translation to hepatocyte transplantation, it is important that hepatocytes maintain their viability and synthetic function after labeling. In this study, primary human hepatocytes were incubated with increasing concentrations of clinical grade SPIOs for different time intervals. SPIOs uptake was confirmed by light and fluorescence microscopy, and intracellular iron content quantified by a colorimetric ferrozine-based assay. Studies were performed to determine if labeling affected cell viability and function. Intracellular iron concentrations increased in a time- and dose-dependent manner after incubation with SPIOs. Labeling had minimal short-term effects on cell attachment and mitochondrial function. However, exposure of hepatocytes to SPIOs resulted in a dose- and time-dependent reduction in protein synthesis. Cell labeling for 16 h had no significant effect on hepatocyte-specific function, but longer periods of incubation resulted in a dose-dependent decrease in albumin production. Hepatocytes incorporated SPIOs at sufficient levels for in vitro detection on a 7-T MRI imaging system, with a minimum of 2,000 SPIO-labeled cells/μl detected by a decreased T2 relaxivity compared to controls. Intrasplenic transplantation of human hepatocytes labeled with 50 μg Fe/ml of SPIOs was performed in nonobese diabetic/severe combined immune deficiency (NOD-Scid) mice. Recipient livers showed a clear decrease in signal intensity on T2*-weighted MR images when compared to controls, allowing detection of hepatocytes. With further experiments to optimize the conditions for labeling human hepatocytes, it should be possible to apply this technique to track hepatocyte transplantation in patients with liver disease.

Key words: Hepatocyte transplantation; Cell labeling; Superapragmagnetic iron oxide nanoparticles (SPIOs); Contrast agents; Cytotoxicity; Magnetic resonance imaging (MRI)

Address correspondence to Dr. Robin D. Hughes, Institute of Liver Studies, King’s College London School of Medicine, Bessemer Road, London SE5 9PJ, UK. Tel: 44 (0) 20 3299 3137; Fax: 44 (0) 20 3299 3760; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it