Cell Transplantation 24(11) Abstracts

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Cell Transplantation, Vol. 24, pp. 2171-2183, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686229
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Review

Regeneration of Degenerated Urinary Sphincter Muscles: Improved Stem Cell-Based Therapies and Novel Imaging Technologies

Bastian Amend,* Martin Vaegler,† Kerstin Fuchs,‡ Julia G. Mannheim,‡ Susanne Will,§ Ulrich Kramer,§ Melanie L. Hart,† Wouter Feitz,¶ Christopher Chapple,# Arnulf Stenzl,* and Wilhelm K. Aicher†

*Department of Urology, University of Tuebingen Hospital, Tuebingen, Germany
†KFO273, Department of Urology, Eberhard Karls University, Tuebingen, Germany
‡Werner Siemens Imaging Center, Department of Preclinical Imaging and Radiopharmacy, Eberhard-Karls University, Tuebingen, Germany
§Department of Diagnostic and Interventional Radiology, University of Tuebingen, Germany
¶Department of Urology, Pediatric Urology, Radboud University Medical Center, Amalia Children’s Hospital, Nijmegen, The Netherlands
#Department of Urology, Royal Hallamshire Hospital, Sheffield Teaching Hospitals NHS Foundation Trust, Sheffield, UK

Stress urinary incontinence (SUI) is a largely ousted but significant medical, social, and economic problem. Surveys suggest that nowadays approximately 10% of the male and 15% of the female population suffer from urinary incontinence at some stage in their lifetime. In women, two major etiologies contribute to SUI: degeneration of the urethral sphincter muscle controlling the closing mechanism of the bladder outflow and changes in lower pelvic organ position associated with degeneration of connective tissue or with mechanical stress, including obesity and load and tissue injury during pregnancy and delivery. In males, the reduction of the sphincter muscle function is sometimes due to surgical interventions as a consequence of prostate cancer treatment, benign prostate hyperplasia, or of neuropathical origin. Accordingly, for women and men different therapies were developed. In some cases, SUI can be treated by physical exercise, electrophysiological stimulation, and pharmacological interventions. If this fails to improve the situation, surgical interventions are required. In standard procedures, endoprostheses for mechanical support of the weakened tissue or mechanical valves for a bladder outflow control are implanted. In 20% of cases treated, repeat procedures are required as implants yield all sorts of side effects in time. Based on preclinical studies, the application of an advanced therapy medicinal product (ATMP) such as implantation of autologous cells may be a curative and long-lasting therapy for SUI. Cellular therapy could also be an option for men suffering from incontinence caused by injury of the nerves controlling the muscular sphincter system. Here we briefly report on human progenitor cells, especially on mesenchymal stromal cells (MSCs), their expansion and differentiation to smooth muscle or striated muscle cells in vitro, labeling of cells for in vivo imaging, concepts of improved, precise, yet gentle application of cells in muscle tissue, and monitoring of injected cells in situ.

Key words: Urology; Stress urinary incontinence (SUI); Stem cell therapy; Stem cell labeling; Stem cell imaging; Advanced therapy medical product (ATMP)

Received May 14, 2014; final acceptance January 7, 2015. Online prepub date: January 20, 2015.
Address correspondence to Wilhelm K. Aicher, KFO273, Department of Urology, UKT, University of Tuebingen, Paul-Ehrlich-Str. 15, 72076 Tuebingen, Germany. Tel: +49 7071 298 7020; Fax: +49 7071 292 5072; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2185-2195, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685113
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Review

Human Induced Pluripotent Stem Cell and Nanotechnology-Based Therapeutics

Wei-Hsiu Liu,*†1 Yuh-Lih Chang,‡§1 Wen-Liang Lo,¶ Hsin-Yang Li,# Chia-Wei Hsiao,** Chi-Hsien Peng,†† Shih-Hwa Chiou,‡‡‡§§ Hsin-I Ma,*† and Shih-Jen Chen‡‡§§

*Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan, ROC
†Department of Neurological Surgery, Tri-Service General Hospital and National Defense Medical Center, Taipei, Taiwan, ROC
‡Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan, ROC
§Department of Pharmacy, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
¶School of Dentistry, National Yang-Ming University and Division of Oral and Maxillofacial Surgery, Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
#Department of Obstetrics and Gynecology, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
**Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
††Department of Ophthalmology, Shin Kong Wu Ho-Su Memorial Hospital and Fu-Jen Catholic University, Taipei, Taiwan, ROC
‡‡School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC
§§Department of Ophthalmology, Taipei Veterans General Hospital, Taipei, Taiwan, ROC

Human induced pluripotent stem cells (hiPSCs) can be genetically reprogrammed to an embryonic stem cell-like state and can provide promising medical applications, such as diagnosis, prognosis, drug screening for therapeutical development, and monitoring disease progression. Despite myriad advances, traditional viral-based reprogramming for generating hiPSCs has safety risks that hinder further practical applications of hiPSCs. In the past decade, nonviral-based reprogramming has been used as an alternative to produce hiPSCs and enhance their differentiation. In addition, the efficiency of nonviral-based reprogramming is generally poor, compared to that of viral-based reprogramming. Recent studies in nanoscale-structured particles have made progress in addressing many applications of hiPSCs for clinical practice. The combination of hiPSCs and nanotechnology will actually act as the therapeutic platform for personalized medicine and can be the remedies against various diseases in the future. In this article, we review recent advances in cellular reprogramming and hiPSC-related research, such as cell source, delivery system, and direct reprogramming, as well as some of its potential clinical applications, including mitochondrial and retinal disease. We also briefly summarize the current incorporation of nanotechnology in patient-specific hiPSCs for future treatments.

Key words: Mesoporous silica nanoparticle; Induced pluripotent stem cells; Cell labeling; DNA delivery

Received March 14, 2014; final acceptance October 3, 2014. Online prepub date: October 8, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Chi-Hsien Peng, Shin Kong Wu Ho-Su Memorial Hospital and Fu-Jen Catholic University, Taipei, Taiwan. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Dr. Shih-Jen Chen, Department of Ophthalmology, Taipei Veterans General Hospital, Taipei 112, Taiwan, ROC. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2197-2208, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688650
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Review

Biological Approaches to Treating Intervertebral Disk Degeneration: Devising Stem Cell Therapies

Inbo Han,*† Alexander E. Ropper,*‡ Deniz Konya,§ Serdar Kabatas,¶ Zafer Toktas,§ Zaid Aljuboori,*# Xiang Zeng,*† John H. Chi,* Ross Zafonte,** and Yang D. Teng*,**,††

*Department of Neurosurgery, Harvard Medical School and Brigham and Women’s Hospital, Boston, MA, USA
†Department of Neurosurgery, CHA University, Sungnam, Korea
‡Department of Spinal Neurosurgery, Barrow Neurological Institute, Phoenix, AZ, USA
§Department of Neurosurgery, Bahcesehir University School of Medicine, Istanbul, Turkey
¶Department of Neurosurgery, Reyap Hospital, Tekirdag, Turkey
#Department of Neurosurgery, School of Medicine, University of Louisville, Louisville, KY, USA
**Department of Physical Medicine and Rehabilitation, Harvard Medical School, Spaulding Rehabilitation Hospital, Boston, MA, USA
††Division of SCI Research, Veterans Affairs Boston Healthcare System, Boston, MA, USA

Intervertebral disk (IVD) degeneration is a common, chronic, and complex degeneration process that frequently leads to back pain and disability, resulting in a major public health issue. In this review we describe biological therapies under preclinical or clinical development with an emphasis on stem cell-based multimodal approaches that target prevention and treatment of IVD degeneration. Systematical review of the basic science and clinical literature was performed to summarize the current status of devising biological approaches to treating IVD degeneration. Since the exact mechanisms underlying IVD degeneration have not yet been fully elucidated and conservative managements appear to be mostly ineffective, current surgical treatment focuses on removal of the pathological disk tissues combined with spinal fusion. The treatment options, however, often produce insufficient efficacy and even serious complications. Therefore, there have been growing demands and endeavors for developing novel regenerative biology-guided strategies for repairing the IVD via delivery of exogenous growth factors, introduction of therapeutic genes, and transplantation of stem cells, or combinatorial therapies. Overall, the data suggest that when applied under a recovery neurobiology principle, multimodal regimens comprising ex vivo engineered stem cell-based disks hold a high potential promise for efficacious clinical translations.

Key words: Intervertebral disk regeneration; Recovery neurobiology; Stem cells; Growth factor; Gene

Received March 11, 2015; final acceptance August 27, 2015. Online prepub date: July 28, 2015.
Address correspondence to Yang D. Teng, 221 Longwood Ave. LM-111A, Boston, MA 02115, USA. Tel: 617-525-8676; Fax: 617-264-5216; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2209-2220, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686201
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Role of Brain-Derived Neurotrophic Factor in Bone Marrow Stromal Cell-Mediated Spinal Cord Repair

Gaby J. Ritfeld,*† Ajay Patel,‡ Alexander Chou,‡ Tabitha L. Novosat,* Deborah G. Castillo,§ Raymund A. C. Roos,† and Martin Oudega*¶#

*Department of Physical Medicine and Rehabilitation, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
†Department of Neurology, Leiden University Medical Center, Leiden, The Netherlands
‡Carnegie Mellon University, Pittsburgh, PA, USA
§Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA
¶Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
#Department of Bioengineering, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

The ability of intraspinal bone marrow stromal cell (BMSC) transplants to elicit repair is thought to result from paracrine effects by secreted trophic factors including brain-derived neurotrophic factor (BDNF). Here we used gene therapy to increase or silence BDNF production in BMSCs to investigate the role of BDNF in BMSC-mediated neuroprotection. In a spinal cord organotypic culture, BMSC-conditioned medium significantly enhanced spinal motoneuron survival by 64% compared with culture medium only. Only conditioned medium of BDNF-hypersecreting BMSCs sustained this neuroprotective effect. In a rat model of spinal cord contusion, a BDNF-dependent neuroprotective effect was confirmed; only with a subacute transplant of BDNF-hypersecreting BMSCs were significantly more spared motoneurons found at 4 weeks postinjury compared with vehicle controls. Spared nervous tissue volume was improved by 68% with both control BMSCs and BDNF-hypersecreting BMSCs. In addition, blood vessel density in the contusion with BDNF-hypersecreting BMSCs was 35% higher compared with BMSC controls and sixfold higher compared with vehicle controls. BDNF-silenced BMSCs did not survive the first week of transplantation, and no neuroprotective effect was found at 4 weeks after transplantation. Together, our data broaden our understanding of the role of BDNF in BMSC-mediated neuroprotection and successfully exploit BDNF dependency to enhance anatomical spinal cord repair.

Key words: Spinal cord injury; Bone marrow stromal cells (BMSCs); Neuroprotection; Blood vessels; Brain-derived neurotrophic factor (BDNF); Gene therapy

Received June 11, 2014; final acceptance December 19, 2014. Online prepub date: January 9, 2015.
Address correspondence to Martin Oudega, Ph.D., University of Pittsburgh School of Medicine, W1452 BSTWR, 200 Lothrop Street, Pittsburgh, PA 15213, USA. Tel/Fax: +1 412-383-6575; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2221-2236, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685122
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Therapeutic Effects of CTLA4Ig Gene-Transduced Adipose Tissue-Derived Mesenchymal Stem Cell Transplantation on Established Autoimmune Thyroiditis

Eun Wha Choi,*† Jung Min Lee,* Hee Woo Lee,‡ Jehoon Yang,* and Hwa Young Youn‡

*Laboratory Animal Research Center, Samsung Biomedical Research Institute, Seoul, Republic of Korea
†School of Medicine, Sungkyunkwan University, Seoul, Republic of Korea
‡Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea

This study aimed to identify the beneficial effects of adipose tissue-derived mesenchymal stem cells (ASCs) and ASCs that overexpress the CTLA4Ig gene (CTLA4Ig-ASCs) on established autoimmune thyroiditis and to examine changes in clinical chemistry parameters and the presence of humoral responses upon repeated long-term administration of autologous ASCs. This study also aimed to acquire desirable results in a preclinical study by using large-sized lab animals and applying ASCs that overexpress therapeutic genes. Experimental autoimmune thyroiditis was induced by immunization with thyroglobulin. Experimental dogs were divided into five groups: (i) ASC IT + IV, (ii) ASC IV, (iii) CTLA4Ig-ASC IT + IV, (iv) CTLA4Ig-ASC IV, and (v) control IT + IV (saline only), and they received intrathyroidal (IT; 10 million cells/250 μl saline per thyroid) administration one time or intravenous (IV; 20 million cells/5 ml) administration seven times within a 101-day period. Blood samples were collected every week, and thyroids were harvested on days 104–106. After serial ASC or CTLA4Ig transplantation, the levels of canine thyroglobulin autoantibodies (TgAA) in serum and the infiltration of T-lymphocytes between the follicles of the thyroid glands were decreased. The expression of FoxP3 in submandibular lymph nodes was significantly increased. Repeated long-term administration of autologous ASCs or CTLA4Ig-ASCs did not generate changes in clinical chemistry parameters or humoral responses.The TgAA test can detect autoimmune thyroiditis years before clinical signs of hypothyroidism occur. Thus, ASC and CTLA4Ig-ASC transplantation in that period can be attractive candidates to ameliorate autoimmune thyroiditis and prevent the development of hypothyroidism.

Key words: CTLA4Ig; Autoimmune disease; Adipose tissue-derived mesenchymal stem cells; Dog; Autoimmune thyroiditis

Received February 2, 2014; final acceptance October 6, 2014. Online prepub date: October 8, 2014.
Address correspondence to Eun Wha Choi, D.V.M., Ph.D., Laboratory Animal Research Center, Samsung Biomedical Research Institute, 81 Irwon-ro, Gangnam-gu, Seoul 135-710, Republic of Korea. Tel: +82-2-3410-3700; Fax: +82-2-3410-1885; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2237-2250, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688894
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Umbilical Cord Blood-Derived Monocytes Improve Cognitive Deficits and Reduce Amyloid-β Pathology in PSAPP Mice

Donna Darlington,*1 Song Li,*¶1 Huayan Hou,*1 Ahsan Habib,* Jun Tian,* Yang Gao,* Jared Ehrhart,§ Paul R. Sanberg,‡ Darrell Sawmiller,* Brian Giunta,*† Takashi Mori,# and Jun Tan*

*Rashid Laboratory for Developmental Neurobiology, Silver Child Development Center, Morsani College of Medicine, University of South Florida, Tampa, FL, USA
†Neuroimmunology Laboratory, Department of Psychiatry and Behavioral Neurosciences, Morsani College of Medicine, University of South Florida, Tampa, FL, USA
‡Center for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, Morsani College of Medicine, University of South Florida, Tampa, FL, USA
§Saneron CCEL Therapeutics Inc., Tampa, FL, USA
¶Center for Translational Research of Neurology Diseases, First Affiliated Hospital, Dalian Medical University, Dalian, China
#Departments of Biomedical Sciences and Pathology, Saitama Medical Center and Saitama Medical University, Kawagoe, Saitama, Japan

Alzheimer’s disease (AD) is the fourth major cause of mortality in the elderly in the US and the leading cause of dementia worldwide. While pharmacological targets have been discovered, there are no true disease-modifying therapies. We have recently discovered that multiple low-dose infusions of human umbilical cord blood cells (HUCBCs) ameliorate cognitive impairments and reduce Aβ-associated neuropathology in PSAPP transgenic mice. However, the mechanism for these effects of HUCBCs remains unclear. In the present study, we examined whether monocytes, as important components of HUCBCs, would have beneficial outcomes on the reduction of AD-like pathology and associated cognitive impairments in PSAPP transgenic AD model mice. PSAPP mice and their wild-type littermates were treated monthly with an infusion of peripheral human umbilical cord blood cell (HUCBC)-derived monocytes over a period of 2 and 4 months, followed by behavioral evaluations, biochemical, and histological analyses. The principal findings of the present study confirmed that monocytes derived from HUCBCs (CB-M) play a central role in HUCBC-mediated cognition-enhancing and Ab pathology-ameliorating activities. Most importantly, we found that compared with CB-M, aged monocytes showed an ineffective phagocytosis of Aβ, while exogenous soluble amyloid precursor protein α (sAPPα) could reverse this deficiency. Pretreating monocytes with sAPPα upregulates Aβ internalization. Our further studies suggested that sAPPα could form a heterodimer with Abs, with the APP672-688 (Aβ1-16) region being responsible for this effect. This in turn promoted binding of these heterodimers to monocyte scavenger receptors and thus promoted enhanced Ab clearance. In summary, our findings suggest an interesting hypothesis that peripheral monocytes contribute to Aβ clearance through heterodimerization of sAPPα with Aβ. Further, declined or impaired sAPPα production, or reduced heterodimerization with Ab, would cause a deficiency in Aβ clearance and thus accelerate the pathogenesis of AD.

Key words: Cord blood; Monocyte; Amyloid β; Soluble amyloid precursor protein α (sAPPα); Heterodimerization

Received June 25, 2015; final acceptance September 08, 2015. Online prepub date: July 30, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Donna Darlington, Rashid Laboratory for Developmental Neurobiology, Silver Child Development Center, Department of Psychiatry and Behavioral Neurosciences, University of South Florida, 3515 E. Fletcher Ave. Tampa, FL 33613-4702, USA. Tel: +1-813-974-5975; Fax: +1-813-974-1130; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2251-2262, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685078
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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A High-Efficiency Induction of Dopaminergic Cells From Human Umbilical Mesenchymal Stem Cells for the Treatment of Hemiparkinsonian Rats

Tsui-Ling Ko,*1 Yu-Yi Fu,† Yang-Hsin Shih,‡§1 Yi-Hui Lin,¶1 Miau-Hwa Ko,#1 Tz-Win Fu,** Tzu-Yung Lin,†† Hsiao-Sheng Hsiao,* Pei-Ming Chu,# and Yu-Show Fu*‡‡

*Institute of Anatomy and Cell Biology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC
†Department of Automation Engineering, Nan-Kai Institute of Technology, Nantou, Taiwan, ROC
‡Department of Neurosurgery, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
§School of Medicine, Taipei Medical University, Taipei, Taiwan, ROC
¶School of Pharmacy, China Medical University, Taichung, Taiwan, ROC
#Department of Anatomy, School of Medicine, China Medical University, Taichung, Taiwan
**Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
††Department of Nursing, Chang Gung University of Science and Technology, Tao-Yuan, Taiwan, ROC
‡‡Department of Education and Research, Taipei City Hospital, Taipei, Taiwan, ROC

The success rate in previous attempts at transforming human umbilical mesenchymal stem cells (HUMSCs) isolated from Wharton’s jelly of the umbilical cord into dopaminergic cells was a mere 12.7%. The present study was therefore initiated to establish a more effective procedure for better yield of dopaminergic cells in such transformation for more effective HUMSC-based therapy for parkinsonism. To examine, in vitro, the effects of enhanced Nurr1 expression in HUMSCs on their differentiation, cells were processed through the three-stage differentiation protocol. The capacity of such cells to synthesize and release dopamine was measured by HPLC. The therapeutic effects of Nurr1-overexppressed HUMSCs were examined in 6-hydroxydopamine-lesioned rats by quantification of rotations in response to amphetamine. Enhanced Nurr1 expression in HUMSCs promoted the transformation into dopaminergic cells in vitro through stepwise culturing in sonic hedgehog, fibroblast growth factor-8, and neuron-conditioned medium. The success rate was about 71%, as determined by immunostaining for tyrosine hydroxylase and around 94 nM dopamine synthesis (intracellular and released into the culture medium), as measured by HPLC. Additionally, transplantation of such cells into the striatum of hemiparkinsonian rats resulted in improvement of their behavioral deficits, as indicated by amphetamine-evoked rotation scores. Viability of the transplanted cells lasted for at least 3 months as verified by positive staining for tyrosine hydroxylase. Nurr1, FGF8, Shh, and NCM can synergistically enhance the differentiation of HUMSCs into dopaminergic cells and may pave the way for HUMSC-based treatments for Parkinson’s disease.

Key words: Human umbilical mesenchymal stem cells (HUMSCs); Nurr1; Dopaminergic cells; Parkinson’s disease (PD); Transplantation

Received April 21, 2014; final acceptance September 16, 2014. Online prepub date: October 6, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Yu-Show Fu, Department of Anatomy, School of Medicine, National Yang-Ming University, No. 155, Sec. 2, Li-Nung Street, Taipei 112, Taiwan, ROC. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Pei-Ming Chu, Department of Anatomy, School of Medicine, China Medical University, No. 91, Hsueh-Shih Road, Taichung 40402, Taiwan, ROC. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2263-2272, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686184
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Surface Coating of Pancreatic Islets With Neural Crest Stem Cells Improves Engraftment and Function After Intraportal Transplantation

Joey Lau,* Svitlana Vasylovska,*† Elena N. Kozlova,† and Per-Ola Carlsson*‡

*Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden
†Department of Neurosciences, Uppsala University, Uppsala, Sweden
‡Department of Medical Sciences, Uppsala University, Uppsala, Sweden

The present study aimed to develop techniques for surface coating of islets with neural crest stem cells (NCSCs) in order to enable cotransplantation to the clinically used liver site and then investigate engraftment and function intraportally of such bioengineered islets. Mouse islets were coated during incubation with enhanced green fluorescent protein (EGFP)-expressing mouse NCSCs and transplanted into the portal vein to cure diabetic mice. An intravenous glucose tolerance test was performed at 1 month posttransplantation. Islet grafts were retrieved and evaluated for vascular density, nerves, and glial cells. NCSCs expressed a vast number of key angiogenic and neurotrophic factors. Mice transplanted with NCSC-bioengineered islets responded better to the glucose load than recipient mice with control islets. NCSCs remained present in the vicinity or had often migrated into the NCSC-coated islets, and an improved islet graft reinnervation and revascularization was observed. Transplanted NCSCs differentiated into both glial and neural cells in the islet grafts. We conclude that bioengineering of islets with NCSCs for intraportal transplantation provides a possibility to improve islet engraftment and function. Pending successful establishment of protocols for expansion of NCSCs from, for example, human skin or bone marrow, this strategy may be applied to clinical islet transplantation.

Key words: Islet transplantation; Stem cells; Neural progenitors; Engraftment; Revascularization; Reinnervation

Received January 21, 2014; final acceptance December 19, 2014. Online prepub date: January 9, 2015.
Address correspondence to Per-Ola Carlsson, M.D., Ph.D., Department of Medical Cell Biology, Uppsala University, Husargatan 3, box 571, SE-75123 Uppsala, Sweden. Tel: +46 18 4714425; Fax: +46 18 4714059; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2273-2283, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686210
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Pharmacological Activation of Nrf2 Pathway Improves Pancreatic Islet Isolation and Transplantation

Shiri Li,* Nosratola D. Vaziri,† Yuichi Masuda,* Mohammad Hajighasemi-Ossareh,* Lourdes Robles,* Aimee Le,* Kelly Vo,* Jefferson Y. Chan,‡ Clarence E. Foster,* Michael J. Stamos,* and HirohitoIchii*

*Department of Surgery, University of California, Irvine, CA, USA
†Department of Medicine, University of California, Irvine, CA, USA
‡Department of Pathology and Laboratory Medicine, University of California, Irvine, CA, USA

Oxidative stress is a major cause of islet damage and loss during the islet isolation process. The Nrf2 pathway plays a critical role in protecting the cells against oxidative stress. The aim of this study was to investigate the effect of an Nrf2 activator (dh404) on islet isolation and transplantation in a rodent model. Islet isolation was conducted using Nrf2-deficient and wild-type mice and vehicle-treated and Nrf2 activator (dh404)-treated rats. Islet yield, viability, and Nrf2 pathway activity were determined. An in vivo islet potency test was done. Islet yield and viability in Nrf2-deficient mice was significantly lower compared to wild-type (p < 0.05) mice. Furthermore, administration of dh404 to normal Sprague–Dawley rats enhanced nuclear translocation of Nrf2 and elevated HO-1 expression in the pancreas. Islet yield and viability in dh404-treated rats was significantly higher compared to the vehicle-treated group (p < 0.05). The diabetes cure rate in nude mice with chemically induced diabetes was significantly greater in those transplanted with islets from the dh404-treated group (6/9) than vehicle-treated rats (2/9, p < 0.05). The Nrf2 pathway plays a significant role in protecting islets against stress caused by the isolation process. Pharmacological activation of the Nrf2 pathway significantly increased HO-1 expression, improved islet yield, viability, and function after transplantation.

Key words: Reactive oxygen species; Oxidative stress; Islet; Isolation; Transplantation; Nuclear erythroid 2-related factor 2 (Nrf2)

Received June 27, 2014; final acceptance December 20, 2014. Online prepub date: January 9, 2015.
Address correspondence to Hirohito Ichii, M.D, Ph.D., Department of Surgery, University of California, Irvine, 333 City Boulevard West, Suite 1205, Orange, CA 92868, USA. Tel: +1 (714) 456-8698; Fax: +1 (714) 456-8796; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2285-2296, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X684060
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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MR Imaging Monitoring of Iron-Labeled Pancreatic Islets in a Small Series of Patients: Islet Fate in Successful, Unsuccessful, and Autotransplantation

Maria Luisa Malosio,*†‡ Antonio Esposito,§¶# Cristina Brigatti,* Anna Palmisano,§¶# Lorenzo Piemonti,*,** Rita Nano,*,** Paola Maffi,*†† Francesco De Cobelli,‡§¶ Alessandro Del Maschio,§¶# and Antonio Secchi#††

*Diabetes Research Institute, San Raffaele Scientific Institute, Milan, Italy
†CNR Institute of Neuroscience, Milan, Italy
‡Humanitas Clinical and Research Center, Rozzano, Milan, Italy
§Radiology Department, San Raffaele Scientific Institute, Milan, Italy
¶Center of Experimental Imaging, San Raffaele Scientific Institute, Milan, Italy
#Vita-Salute San Raffaele University, Milan, Italy
**Human Islet Isolation and Transplantation Program, Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy
††Transplant Medicine Unit, San Raffaele Scientific Institute, Milan, Italy

Islet transplantation is one of the most promising and effective therapies for restoring normoglycemia in type 1 diabetes (T1D) patients, but islet engraftment is one of the main obstacles hampering long-term success. Monitoring graft loss, caused either by immunological or nonimmunological events, occurring in the first phase after transplantation and at later stages of a patient’s life is a very important issue. Among the imaging approaches previously applied, magnetic resonance imaging (MRI) monitoring of islet fate following labeling with superparamagnetic iron oxide agents yielded promising results. The aim of this study was to translate into patients the method of islet labeling and MRI monitoring developed in our preclinical setting and to compare imaging results with graft clinical outcome. Three T1D patients and one nondiabetic patient undergoing autotransplantation following subtotal pancreatectomy received Endorem®-labeled islets. Patients were monitored by MRI and metabolically (HbA1c, exogenous insulin requirement, and C-peptide, TEF) at 1, 3, and 7 days following transplantation and once a month up to 10 months. Labeled transplanted islets appeared as hypointense areas scattered within the liver parenchyma, whose absolute number at 24 h after transplantation reflected the labeling efficiency. In patients #1 and #3 with good midterm graft function, MRI follow-up showed an important early loss of hypointense spots followed by a slow and progressive disappearance at later timepoints. Graft loss of function in patient #2 4 weeks after transplantation was associated with the complete disappearance of all hypointense signals. The autotransplanted patient, stably insulin free, showed no significant signal reduction during the first 3 days, followed by loss of spots similar to a patient with good midterm graft function. These results suggest that MRI monitoring of islet transplantation at early time points could represent a meaningful readout for helping in predicting transplant failure or success, but its relevance for mid/long-term islet function assessment appears evanescent.

Key words: Pancreatic islet transplantation; Type 1 diabetes patients; Autoimmunity; Autologous islet transplantation; Superparamagnetic iron oxide particles (SPIO); Magnetic resonance imaging (MRI); Inflammation; Cytokines

Received January 14, 2014; final acceptance August 22, 2014. Online prepub date: August 29, 2014.
Address correspondence to Maria Luisa Malosio, Ph.D., CNR Institute of Neuroscience - Via Vanvitelli 32, 20129 Milan, Italy. Tel: +390282245254; Fax: +390282245290; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2297-2305, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685636
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Regional Implantation of Autologous Adipose Tissue-Derived Cells Induces a Prompt Healing of Long-Lasting Indolent Digital Ulcers in Patients With Systemic Sclerosis

Nicoletta Del Papa,* Gabriele Di Luca,† Domenico Sambataro,* Eleonora Zaccara,* Wanda Maglione,* Armando Gabrielli,‡ Paolo Fraticelli,‡ Gianluca Moroncini,‡ Lorenzo Beretta,§ AlessandroSantaniello,§ Gianluca Sambataro,¶ Roberto Ferraresi,# and Claudio Vitali**

*U.O.C. Day Hospital Reumatologia, Ospedale G. Pini, Milano, Italy
†U.O.S. Chirurgia Vascolare, Ospedale G. Pini, Milano, Italy
‡U.O. Clinica Medica, Dipartimento di Medicina Interna, Ospedali Riuniti, Ancona, Italy
§Centro di Riferimento per le Malattie Autoimmuni Sistemiche, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico di Milano, Milano, Italy
¶Dipartimento Medicina Interna e Reumatologia, Campus Biomedico, Roma, Italy
#Laboratorio Emodinamica Periferica Interventistica, Humanitas Gavazzeni, Bergamo, Italy
**Servizio di Reumatologia, Istituto San Giuseppe, Anzano del Parco, Como, Italy

Digital ulcers (DUs) are a rather frequent and invalidating complication in systemic sclerosis (SSc), often showing a very slow or null tendency to heal, in spite of the commonly used systemic and local therapeutic procedures. Recently, stem cell therapy has emerged as a new approach to accelerate wound healing. In the present study, we have tentatively treated long-lasting and poorly responsive to traditional therapy SSc-related DUs by implantation of autologous adipose tissue-derived cell (ATDC) fractions. Fifteen patients with SSc having a long-lasting DU in only one fingertip who were unresponsive to intensive systemic and local treatment were enrolled in the study. The grafting procedure consisted of the injection, at the basis of the corresponding finger, of 0.5–1 ml of autologous ATDC fractions, separated by centrifugation of adipose tissue collected through liposuction from subcutaneous abdominal fat. Time to heal after the procedure was the primary end point of the study, while reduction of pain intensity and of analgesic consumption represented a secondary end point. Furthermore, the posttherapy variation of the number of capillaries, observed in the nailfold video capillaroscopy (NVC) exam and of the resistivity in the digit arteries, measured by high-resolutionechocolor-Doppler, were also taken into account. A rather fast healing of the DUs was reached in all of the enrolled patients (mean time to healing 4.23 weeks; range 2–7 weeks). A significant reduction of pain intensity was observed after a few weeks (p < 0.001), while the number of capillaries was significantly increased at 3- and 6-month NVC assessment (p < 0.0001 in both cases). Finally, a significant after-treatment reduction of digit artery resistivity was also recorded (p < 0.0001). Even with the limitations related to the small number of patients included and to the open-label design of the study, the observed strongly favorable outcome suggests that local grafting with ATDCs could represent a promising option for the treatment of SSc-related DUs unresponsive to more consolidated therapies.

Key words: Systemic sclerosis; Adipose tissue; Stem cells; Autologous graft; Ulcers

Received July 26, 2014; final acceptance November 11, 2014. Online prepub date: December 12, 2014.
Address correspondence to Nicoletta Del Papa, M.D., U.O.C. Day Hospital Reumatologia, Ospedale G. Pini, Via Pini, 9 - 20122 Milano, Italy. Tel: +39-0258296663; Fax: +39-0258296495; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2307-2322, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685771
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Cardiosphere Conditioned Media Influence the Plasticity of Human Mediastinal Adipose Tissue-Derived Mesenchymal Stem Cells

Camilla Siciliano,*† Isotta Chimenti,* Mohsen Ibrahim,‡ Chiara Napoletano,§ Giorgio Mangino,* Gaia Scafetta,* Giuseppe Biondi Zoccai,* Erino Angelo Rendina,‡ Antonella Calogero,* Giacomo Frati,*¶1and Elena De Falco*1

*Department of Medical-Surgical Sciences and Biotechnologies, Sapienza University of Rome, Latina, Italy
†Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy
‡Division of Thoracic Surgery, Department of Medical-Surgical Science and Translational Medicine, Sapienza University of Rome, S. Andrea Hospital, Rome, Italy
§Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy
¶Department of AngioCardioNeurology, IRCCS Neuromed, Pozzilli, Italy

Nowadays, cardiac regenerative medicine is facing many limitations because of the complexity to find the most suitable stem cell source and to understand the regenerative mechanisms involved. Mesenchymal stem cells (MSCs) have shown great regenerative potential due to their intrinsic properties and ability to restore cardiac functionality, directly by transdifferentiation and indirectly by paracrine effects. Yet, how MSCs could respond to definite cardiac-committing microenvironments, such as that created by resident cardiac progenitor cells in the form of cardiospheres (CSs), has never been addressed. Recently, a putative MSC pool has been described in the mediastinal fat (hmADMSCs), but both its biology and function remain hitherto unexplored. Accordingly, we investigated the potential of hmADMSCs to be committed toward a cardiovascular lineage after preconditioning with CS-conditioned media (CCM). Results indicated that CCM affects cell proliferation. Gene expression levels of multiple cardiovascular and stemness markers (MHC, KDR, Nkx2.5, Thy-1, c-kit, SMA) are significantly modulated, and the percentage of hmADMSCs preconditioned with CCM and positive for Nkx2.5, MHC, and KDR is significantly higher relative to FBS and explant-derived cell conditioned media (EDCM, the unselected stage before CS formation). Growth factor-specific and survival signaling pathways (i.e., Erk1/2, Akt, p38, mTOR, p53) present in CCM are all equally regulated. Nonetheless, earlier BAD phosphorylation (Ser112) occurs associated with the CS microenvironment (and to a lesser extent to EDCM), whereas faster phosphorylation of PRAS40 in FBS, and of Akt (Ser473) in EDCM and 5-azacytidine occurs compared to CCM. For the first time, we demonstrated that the MSC pool held in the mediastinal fat is adequately plastic to partially differentiate in vitro toward a cardiac-like lineage. Besides, we have provided novel evidence of the potent inductive niche-like microenvironment that the CS structure can reproduce in vitro. hmADMSCs can represent an interesting tool in order to exploit their possible role in cardiovascular diseases and treatment.

Key words: Adipose tissue-derived mesenchymal stem cells; Cardiospheres (CSs); Cardiac-like differentiation; Conditioned media; 5-Azacytidine

Received June 20, 2014; final acceptance December 11, 2014. Online prepub date: January 8, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Elena De Falco, Ph.D., Assistant Professor, Sapienza University of Rome, Faculty of Pharmacy and Medicine, Department of Medical-Surgical Sciences and Biotechnologies, C.so della Repubblica 79, 04100 Latina, Italy. Tel: +39-07731757234; Fax: +39-0773175754; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2323-2335, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X684051
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

ASC Spheroid Geometry and Culture Oxygenation Differentially Impact Induction of Preangiogenic Behaviors in Endothelial Cells

Matthew L. Skiles,*† Brandon Hanna,† Lindsay Rucker,† Allison Tipton,† Aidan Brougham-Cook,† Ehsan Jabbarzadeh,*† and James O. Blanchette*†

*Chemical Engineering Department, University of South Carolina, Columbia, SC, USA
†Biomedical Engineering Program, University of South Carolina, Columbia, SC, USA

Cell-based angiogenic therapies offer potential for the repair of ischemic injuries, while avoiding several of the limitations associated with material-based growth factor delivery strategies. Evidence supports that applying MSCs as spheroids rather than dispersed cells can improve retention and enhance therapeutic effect through increased secretion of angiogenic factors due to hypoxia. However, while spheroid culture appears to modulate MSC behavior, there has been little investigation of how major culture parameters that affect cellular oxygen tension, such as external oxygenation and culture size, impact the angiogenic potential of spheroids. We cultured equal numbers of adipose-derived stem cells (ASCs) as spheroids containing 10,000 (10k) or 60,000 (60k) cells each, in 20% and 2% oxygen. VEGF secretion varied among the sample groups, with 10k, 2% O2 spheroids exhibiting the highest production. Spheroid-conditioned media was applied to HUVEC monolayers, and proliferation was assessed. Spheroids of either size in 2% oxygen induced comparable proliferation compared to a 2 ng/ml VEGF control sample, while spheroids in 20% oxygen induced less proliferation. Spheroids were also applied in coculture with HUVEC monolayers, and induction of migration through a Transwell membrane was evaluated. Sixty thousand, 2% O2 spheroids induced similar levels of migration as VEGF controls, while 10k, 2% O2 spheroids induced significantly more. Ten thousand, 20% spheroids performed no better than VEGF-free controls. We conclude that the therapeutic ability of ASC spheroids to stimulate angiogenesis in endothelial cells is affected by both culture size and oxygenation parameters, suggesting that, while ASC spheroids offer potential in the treatment of injured and ischemic tissues, careful consideration of culture size in respect to in vivo local oxygen tension will be necessary for optimal results.

Key words: Cell-based delivery; Angiogenesis; Adipose-derived stem cells (ASCs); Spheroid culture; Hypoxia

Received March 11, 2014; final acceptance August 19, 2014. Online prepub date: August 21, 2014.
Address correspondence to Dr. James O. Blanchette, 208 Sesqui Trail, Columbia, SC 29223, USA. Tel: +1-803-777-1541; Fax: +1-803-777-0973; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2337-2351, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685780
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adipose-Derived Stem Cells From Both Visceral and Subcutaneous Fat Deposits Significantly Improve Contractile Function of Infarcted Rat Hearts

Chao Chi,*†‡1 Fei Wang,*†‡1 Bo Xiang,‡ Jixian Deng,†‡ Shangdian Liu,* Hung-Yu Lin,† Kanmani Natarajan,§ Gang Li,* Lei Wang,* Jian Wang,§ Francis Lin,¶ Darren H. Freed,# Rakesh C. Arora,**Hongyu Liu,* and Ganghong Tian†‡

*Department of Cardiac Surgery, First Affiliated Hospital, Harbin Medical University, Harbin, China
†National Research Council, Winnipeg, Manitoba, Canada
‡Department of Physiology and Pathophysiology, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada
§Department of Vascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Hubei, China
¶Department of Physics and Astronomy, University of Manitoba, Winnipeg, Manitoba, Canada
#University of Alberta, Edmonton, Canada
**St. Boniface Research Centre, Winnipeg, Manitoba, Canada

Adipose-derived stem cells (ASCs) from subcutaneous and visceral adipose tissues have been studied individually. No studies have compared their abilities in treatment of heart failure. This study was designed to evaluate whether ASCs from the two sources could provide a long-term improvement of cardiac function in infarcted hearts. Rat subcutaneous and visceral adipose tissues were excised for isolation of ASCs. Morphology, yield, proliferation, surface markers, differentiation, and cytokine secretion of the subcutaneous ASCs (S-ASCs) and visceral ASCs (V-ASCs) were analyzed. Then a rat model of myocardial infarction (MI) was established by a coronary occlusion. Seven days after occlusion, S-ASCs (n = 22), V-ASCs (n = 22), and Dulbecco’s modified Eagle medium (DMEM, n = 20) were injected into the infarct rim, respectively. Cardiac function was then monitored with MRI for up to 6 months. The hearts were then removed for histological assessments. The yield of V-ASCs per gram of the visceral adipose depot was significantly greater than that of S-ASCs in 1 g of the subcutaneous adipose depot. On the other hand, the S-ASCs showed a greater proliferation rate and colony-forming unit relative to the V-ASCs. In addition, the infarcted hearts treated with either S-ASCs or V-ASCs showed a significantly greater left ventricular ejection fraction (LVEF) than those treated with DMEM at 4 weeks and 6 months following the cell/DMEM transplantation. Moreover, the infarct sizes of both S-ASC and V-ASC-treated hearts were significantly smaller than that in the DMEM-treated hearts. MRI showed the implanted ASCs at the end of 6 months of recovery. Despite the differences in cell yield, proliferation, and colony formation capacity, both S-ASCs and V-ASCs provide a long-lasting improvement of cardiac contractile function in infarcted hearts. We conclude that the subcutaneous and visceral adipose tissues are equally effective cell sources for cell therapy of heart failure.

Key words: Subcutaneous adipose-derived stem cells; Visceral adipose-derived stem cells; Myocardial infarction; Heart failure; Magnetic resonance imaging

Received July 11, 2014; final acceptance December 16, 2014. Online prepub date: January 5, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Hongyu Liu, Department of Cardiac Surgery, First Affiliated Hospital, Harbin Medical University, 23 Youzheng St, Nangang, Harbin, Heilongjiang 150001, China. Tel: +8645185555817; Fax: +8645185555817; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Dr.Ganghong Tian, National Research Council Canada, 435 Ellice Ave, Winnipeg, Manitoba, Canada R3B, 1Y6. Tel: +1 (204) 984-6654; Fax: +1 (204) 984-7036;

E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2353-2366, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685753
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

DFO Enhances the Targeting of CD34-Positive Cells and Improves Neovascularization

Zijing Du,1 Tao Zan,1 Xiaolu Huang, Lingling Sheng, Haizhou Li, Hua Li, and Qingfeng Li

Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, P.R. China

Desferrioxamine (DFO), an iron chelator, mimicked hypoxia by inhibiting HIF-1α degradation and upregulated angiogenic factors. In this experiment, we elucidated the effect of DFO on CD34-positive cell migration and neovascularization. CD34-positive cells were cultured in media with DFO or an inhibitor and subjected to in vitro tubule formation and the expression of factors. Nude mice were randomly divided into five groups of 12: control, CD34, CD34-DFO, CD34-DFO-AMD (AMD3100, CXCR4 inhibitor), and CD34-DFO-LY (LY294002, the PI3K inhibitor) groups. Limb perfusion and in vivo imaging was evaluated by laser speckle imaging (LSI) and bioluminescence imaging (BLI). Capillary density was examined 14 days after surgery, and the relevant mechanism was also explored. In vitro, DFO significantly increased the tube formation and expression of angiogenic factors in CD34-positive cells, which were blocked by the PI3K inhibitor, LY294002. DFO enhanced blood flow, the function of the ischemic hindlimb, and the levels of VEGF. Further, p-eNOS and p-Akt increased in response to the ischemia. BLI showed that DFO increased the number of CD34-positive cells targeted to the ischemic sites. Immunohistofluorescence revealed that the capillary density in the ischemic hindlimb was significantly higher in the DFO treatment group compared with the other groups. However, all of these effects were diminished by LY294002. DFO treatment enhanced CD34-positive cell targeting and improved neovascularization via the PI3K/Akt signal transduction pathway in an ischemic hindlimb.

Key words: Iron chelator; Vasculogenesis; Angiogenesis; PI3K/AKT; HIF-1α

Received July 12, 2014; final acceptance December 4, 2014. Online prepub date: December 12, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Qingfeng Li, M.D., Ph.D., Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizhaoju Road, Shanghai, P.R. China 200011. Fax: +86-21-63089567; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2367-2377, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685645
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adipose Tissue-Derived Mesenchymal Stem Cells Induce Expansion of Interleukin-10-Producing Regulatory B Cells and Ameliorate Autoimmunity in a Murine Model of Systemic Lupus Erythematosus

Min-Jung Park,*1 Seung-Ki Kwok,*†1 Sung-Hee Lee,* Eun-Kyung Kim,* Sung-Hwan Park,*†2 and Mi-La Cho*2

*The Rheumatism Research Center, The Catholic University of Korea, Seoul, South Korea
†Division of Rheumatology, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, South Korea

Mesenchymal stem cells (MSCs) are multipotent cells characterized by immunomodulatory properties and are therefore considered a promising tool for the treatment of autoimmune diseases. One functional B-cell subset, regulatory B cells (Bregs), has recently been shown to restrain excessive inflammatory responses in autoimmune diseases. In the present study, we investigated the impact of human adipose-derived MSCs on Bregs and their therapeutic effect in an animal model of systemic lupus erythematosus (SLE). Coculture of human adipose-derived MSCs with splenocytes from C57BL/6 mice expanded the population of interleukin-10-producing B cells (B10 B cells). In vivo treatment with human adipose-derived MSCs reduced serum anti-double-stranded antibody levels and improved renal pathology of lupus mice (Roquinsan/san mice). MSCs decreased ICOS+CD44+ follicular helper T cells, Th1 cells and Th17 cells, in spleens of Roquinsan/san mice. In contrast, MSCs increased Foxp3-expressing regulatory T cells. MSCs also decreased the size and number of germinal centers and effector B cells. As expected, in vivo treatment with MSCs expanded the population of Bregs in spleens of Roquinsan/san mice. Our results indicate that human adipose-derived MSCs induce the expansion of Bregs and ameliorate autoimmunity in a murine model of SLE. These findings suggest that human adipose-derived MSCs may be a promising therapeutic strategy targeting B-cell-mediated autoimmune diseases such as SLE.

Key words: Mesenchymal stem cells (MSCs); Regulatory B cells (Bregs); Systemic lupus erythematosus (SLE); Follicular helper T cells (Tfh cells); Interleukin (IL)-10

Received November 12, 2013; final acceptance November 17, 2014. Online prepub date: December 12, 2014.
1These authors provided equal contribution to this work.
2These authors provided equal contribution to this work.
Address correspondence to Mi-La Cho, Ph.D., Rheumatism Research Center, Catholic Institutes of Medical Science, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul 137-040, South Korea. Tel: +82-2-2258-7467; Fax: +82-2-599-4287; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Sung- Hwan Park, M.D., Ph.D., Division of Rheumatology, Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul St. Mary’s Hospital, 505 Banpo-dong, Seocho-gu, Seoul 137-701, South Korea. Tel: +82 2 2258 6011; Fax: +82 2 599 3589; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2379-2390, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685654
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

In Vivo Tracking and Fate of Intra-Articularly Injected Superparamagnetic Iron Oxide Particle-Labeled Multipotent Stromal Cells in an Ovine Model of Osteoarthritis

Uta Delling,* Walter Brehm,* Marco Metzger,† Eberhard Ludewig,‡ Karsten Winter,§ and Henriette Julke§

*University of Leipzig, Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, Leipzig, Germany
†Fraunhofer IGB Project Group Oncology, Department for Tissue Engineering and Regenerative Medicine, Wuerzburg, Germany
‡University of Leipzig, Faculty of Veterinary Medicine, Department of Small Animal Medicine, Leipzig, Germany
§University of Leipzig, Translational Centre for Regenerative Medicine (TRM), Leipzig, Germany

In this study, superparamagnetic iron oxide (SPIO) particle-labeled mesenchymal stromal cells (MSCs) were injected intra-articularly into osteoarthritic knee joints. Their fate and distribution were evaluated using magnetic resonance imaging (MRI) and macroscopic and histologic postmortem examination. Osteoarthritis was induced in 12 sheep by bilateral meniscectomy. After 6 weeks, one knee joint received 10 × 106 SPIO-labeled MSCs (Molday Ion Rhodamine B). Contralateral knees received a control injection of a) PBS, b) SPIO in PBS, c) 10 × 106 nonvital SPIO-labeled MSCs in PBS, or d) no injection. MR images were acquired immediately after injection and 1, 4, 8, and 12 weeks thereafter using a 0.5-T unit and a T2* sequence. Signal intensity of synovial fluid and synovial lining was assessed semiquantitatively using a scoring system. Viable SPIO-labeled MSCs produced a strong hypointense signal in the synovial fluid immediately after injection, but normal signal intensity of the synovial fluid was observed 1 week later. Synovial lining maintained its hypointensity throughout the study period. Nonvital SPIO-labeled MSCs induced hypointense signals of the synovial fluid; synovial lining appeared weak and inconsistently hypointense in the following weeks. Pure SPIO produced a strong hyperintense signal in the synovial fluid at the time of injection only. Histologically, in all knee joints receiving viable SPIO-labeled MSCs, SPIO particles were detected (Prussian blue) within the synovial lining, dorsal fat pad, and neomeniscus tissue, but not in osteochondral samples. Few SPIO particles were detected in joints injected with nonvital SPIO-labeled MSCs. Immunohistologically, no increased cell death (TUNEL) was observed in the area of detected SPIO particles, but we did observe potential chondrogenic cell differentiation (Safranin O or S100b). We conclude that viable SPIO-labeled MSCs remain detectable within the joint for 12 weeks and attach themselves to some but not all diseased joint structures.

Key words: Multipotent stromal cells (MSCs); Cell tracking; In vivo; Superparamagnetic iron oxide (SPIO) particles; Osteoarthritis (OA)

Received July 9, 2014; final acceptance November 20, 2014. Online prepub date: December 12, 2014.
Address correspondence to Uta Delling, University of Leipzig, Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, An den Tierkliniken 21, 04103 Leipzig, Germany. Tel: +49 341 9738253; Fax: +49 341 97398269; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2391-2407, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686193
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Longitudinal Evaluation of Effects of Intra-Articular Mesenchymal Stromal Cell Administration for the Treatment of Osteoarthritis in an Ovine Model

Uta Delling,* Walter Brehm,* Eberhard Ludewig,† Karsten Winter,‡ and Henriette Jülke‡

*University of Leipzig, Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, Leipzig, Germany
†University of Leipzig, Faculty of Veterinary Medicine, Department of Small Animal Medicine, Leipzig, Germany
‡University of Leipzig, Translational Centre for Regenerative Medicine (TRM), Leipzig, Germany

In this study, the therapeutic effect of intra-articularly injected autologous mesenchymal stromal cells (MSCs) was evaluated in an ovine osteoarthritis (OA) model using consecutive magnetic resonance imaging (MRI), radiography, and macroscopic and histologic postmortem examination. In 12 sheep, OA was induced by bilateral, lateral meniscectomy. After 6 weeks, 20 × 106 bone marrow-derived MSCs (50% MSCs were superparamagnetic iron oxide particle labeled) were injected intra-articularly into one knee joint. The contralateral knee served as negative control. MR images were acquired before OA induction, immediately before and after MSC injection, and 1, 4, 8, and 12 weeks thereafter using a 0.5T unit and a T2* gradient echo sequence. Radiographs were obtained before OA induction, at MSC injection, and 12 weeks thereafter. The MRI scoring system included articular cartilage, bone, joint capsule, and synovial fluid evaluation. The radiographic scoring system included the joint space and bone. Postmortem evaluation entailed macroscopic and histologic assessment. Longitudinal MRI revealed a significant deceleration of OA progression in MSC-treated joints. However, at the conclusion of the study, there was no significant difference in the degree of OA detected by MRI, radiography, and postmortem evaluation between the treatment and control group. The degree of OA on MRI varied among the 12 animals at the time of injection, but there was no difference between the left and right limb. In conclusion, intra-articular MSCs decreased OA progression. However, no significant treatment effects were seen at the conclusion of the study at 12 weeks. This somewhat contradicts previously published results. Nevertheless, the choice of OA model, outcome measures, or lack of additional medication might explain the differences. Our results indicate that OA might benefit from intra-articular MSC injection, but further studies are needed to refine patient selection and injection parameters for a more substantially improved outcome.

Key words: Mesenchymal stromal cells (MSCs); Osteoarthritis (OA); Magnetic resonance imaging (MRI); Ovine model; Osteoarthritis Research Society International (OARSI)

Received August 5, 2014; final acceptance December 21, 2014. Online prepub date: January 9, 2015.
Address correspondence to Uta Delling, University of Leipzig, Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, An den Tierkliniken 21, 04103 Leipzig, Germany. Tel: +49 341 9738253; Fax: +49 341 97398269; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2409-2422, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681577
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Impact of Short-Term Liquid Storage on Human CD133+ Stem Cells

Cornelia A. Lux,1 Peter Mark,1 Christian Klopsch, Michael Laupheimer, Hoang Tu-Rapp, Wenzhong Li, Nan Ma, Gustav Steinhoff, and Robert David

Reference and Translation Center for Cardiac Stem Cell Therapy, University of Rostock, Rostock, Germany

Stem cell transplantation is a viable strategy for regenerative medicine. However, it is inevitable to have cells undergo storage for several hours or days due to processing and transportation. Therefore, it is crucial to have rigidly controlled conditions ensuring the therapeutic benefit of isolated stem cells. In the present study, we investigated the impact of short-term storage on human CD133+ cells. CD133+ cells were isolated from human bone marrow and kept at standardized nonfreezing storage conditions for up to 72 h. Cell viability (apoptosis/necrosis) and expression of CD133 and CXCR4 were analyzed by flow cytometry. Metabolic activity was determined using an MTT assay; colony-forming ability, as well as endothelial-like differentiation, was further evaluated. A qRT-PCR array was employed to investigate the expression of stemness genes. CD133 and CXCR4 expressions were preserved at all time points. After 30 h, cell number and metabolic activity decreased, although no significant changes were detected in cell viability and proliferation as well as endothelial-like differentiation. Cell viability and proliferation decreased significantly only after 72 h of storage. Our results indicate that storage of isolated human CD133+ bone marrow stem cells in liquid allows for high viability and functionality. However, storage time should be limited in order to avoid cell loss.

Key words: Hematopoietic stem cells; Transplantation; Cardiac regeneration; CD133; Preservation

Received March 23, 2012; final acceptance April 22, 2014. Online prepub date: May 2, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Cornelia A. Lux, RTC, BMFZ Schillingallee 68, 18057 Rostock, Germany. Tel: +0049-381-494-3904; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it