Cell Transplantation 24(8) Abstracts

Return to Cell Transplantation>

Cell Transplantation, Vol. 24, pp. 1405-1422, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682431
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Therapeutic Implications of Newly Identified Stem Cell Populations From the Skin Dermis

Zelin Chen, Yu Wang, and Chunmeng Shi

Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing, China

Skin, the largest organ of the body, is a promising reservoir for adult stem cells. The epidermal stem cells and hair follicle stem cells have been well studied for their important roles in homeostasis, regeneration, and repair of the epidermis and appendages for decades. However, stem cells residing in dermis were not identified until the year 2001, when a variety of stem cell subpopulations have been isolated and identified from the dermis of mammalian skin such as neural crest stem cells, mesenchymal stem cell-like dermal stem cells, and dermal hematopoietic cells. These stem cell subpopulations exhibited capabilities of self-renewing, multipotent differentiating, and immunosuppressive properties. Hence, the dermis-derived stem cells showed extensive potential applications in regenerative medicine, especially for wound healing/tissue repair, neural repair, and hematopoietic recovery. Here we summarized current research on the stem cell subpopulations derived from the dermis and aimed to provide a comprehensive review on their isolation, specific markers, differentiation capacity, and the functional activities in homeostasis, regeneration, and tissue repair.

Key words: Dermis; Neural crest stem cells; Mesenchymal stem cells (MSCs); Hematopoietic cells

Received January 24, 2014; final acceptance June 24, 2014. Online prepub date: June 26, 2014.
Address correspondence to Chunmeng Shi, Ph.D., Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing 400038, China. Tel: +86-023-68752280; Fax: +86-23-68752009; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Yu Wang, Ph.D., Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing 400038, China. Tel: +86-023-68771726; Fax: +86-23-68752009; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1423-1429, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683052
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Tumorigenicity Testing in Athymic Mice of Cultured Human Melanocytes for Transplantation in Engineered Skin Substitutes

Steven T. Boyce,*† Rachel L. Zimmerman,* and Dorothy M. Supp*†

*Department of Surgery, University of Cincinnati College of Medicine, Cincinnati, OH, USA
†Research Department, Shriners Hospitals for Children, Cincinnati, OH, USA

Autologous engineered skin substitutes (ESS) have been shown to close excised, full-thickness burns, but are consistently hypopigmented due to depletion of human melanocytes (hM) during culture of keratinocytes. Hypothetically, addition of hM to ESS may restore uniform pigmentation, but may also promote neoplasia and tumor formation. To evaluate this risk, 16 strains of hM were isolated and propagated in selective culture medium, then injected subcutaneously into athymic mice (1 × 107
hM/animal; n = 6/strain) and followed for 24 weeks. Human melanoma cells (SK-Mel-2, SK-Mel-5) served as positive controls. No detectable tumors formed from hM strains derived from normal skin. In contrast, SK-Mel-2 formed tumors in 50% of mice, and SK-Mel-5 formed tumors in 83% of mice. Histopathology confirmed the tumorigenic anatomy of the controls and the presence of hM that were not tumorigenic in the test groups. These results support the safety of cultured hM for transplantation to restore uniform skin pigmentation in wounds closed with ESS.

Key words: Hypopigmentation; Tumorigenicity; Cultured melanocytes; Transplantation; Engineered skin; Regenerative medicine

Received January 28, 2014; final acceptance July 22, 2014. Online prepub date: July 23, 2014.
Address correspondence to Steven Boyce, Ph.D., Department of Surgery, University of Cincinnati College of Medicine, P.O. Box 670558, Cincinnati, OH 45267-0558, USA. Tel: +1-513-872-6080; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1432-1450, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682107
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Nuclear Localization Signal-Enhanced Polyurethane-Short Branch Polyethylenimine-Mediated Delivery of Let-7a Inhibited Cancer Stem-Like Properties by Targeting the 3-UTR of HMGA2 in Anaplastic Astrocytoma

Meng-Yin Yang,*†‡ Ming-Teh Chen,§¶1 Pin-I Huang,§#1 Chien-Ying Wang,§** Yun-Chin Chang,§†† Yi-Ping Yang,§†† Wen-Liang Lo,§†† Wen-Hsing Sung,§‡‡ Yi-Wen Liao,§†† Yi-Yen Lee,§¶ Yuh-LihChang,§†† Ling-Ming Tseng,§** Yi-Wei Chen,§# and Hsin-I Ma*§§

*Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan
†Department of Neurological Surgery, Taichung Veterans General Hospital, Taichung, Taiwan
‡Department of Neurological Surgery, Jan-Ai Hospital, Taichung City, Taiwan
§School of Medicine, National Yang-Ming University, Taipei, Taiwan
¶Department of Neurosurgery, Neurological Institute, Taipei Veterans General Hospital, Taipei, Taiwan
#Cancer Center, Taipei Veterans General Hospital, Taipei, Taiwan
**Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
††Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan
‡‡Department of Neurosurgery, Cheng-Hsin General Hospital, Taipei, Taiwan
§§Department of Neurological Surgery, Tri-Service General Hospital and National Defense Medical Center, Taipei, Taiwan

Anaplastic astrocytoma (AA) is a grade III glioma that often occurs in middle-aged patients and presents a uniformly poor prognosis. A small subpopulation of cancer stem cells (CSCs) possessing a self-renewing capacity is reported to be responsible for tumor recurrence and therapeutic resistance. An accumulating amount of microRNAs (miRNA) were found aberrantly expressed in human cancers and regulate CSCs. Efforts have been made to couple miRNAs with nonviral gene delivery approaches to target specific genes in cancer cells. However, the efficiency of delivery of miRNAs to AA-derived CSCs is still an applicability hurdle. The present study aimed to investigate the effectiveness and applicability of nonviral vector-mediated delivery of Let-7a with regard to eradication of AA and AA-derived CSC cells. Herein, our miRNA/mRNA microarray and RT-PCR analysis showed that the expression of Let-7a, a tumor-suppressive miRNA, is inversely correlated with the levels of HMGA2 and Sox2 in the AA side population (SP+) cells. Luciferase reporter assay showed that Let-7a directly targets the 3′-UTRs of HMGA2 in AA-SP+
cells. Knockdown of HMGA2 significantly suppressed the protein expression of Sox2 in AA-SP+ cells, whereas overexpression of HMGA2 upregulated Sox2 expression in AA-SP. Nuclear localization signal (NLS) peptides can facilitate nuclear targeting of DNA and are used to improve gene delivery. Using polyurethane-short branch polyethylenimine (PU-PEI) as a therapeutic delivery vehicle, we conjugated NLS with Let-7 and successfully delivered it to AA-SP+ cells, resulting in significantly suppressed expression of HMGA2 and Sox2, tumorigenicity, and CSC-like abilities. This treatment facilitated the differentiation of AA-SP+ cells into non-SP CSCs. Furthermore, PU-PEI-mediated delivery of NLS-conjugated Let-7a in AA-SP+ cells suppressed the expression of drug-resistant and antiapoptotic genes, and increased cell sensitivity to radiation. Finally, the in vivo delivery of PU-PEI-NLS-Let-7a significantly suppressed the tumorigenesis of AA-SP+ cells and synergistically improved the survival rate of orthotopically AA-SP+-transplanted immunocompromised mice when combined with radiotherapy. Therefore, PU-PEI-NLS-Let-7a is a potential novel therapeutic approach for AA.

Key words: MicroRNA (miRNA); Nuclear localization signal (NLS); Nonviral vector; Anaplastic astrocytoma (AA)

Received January 3, 2014; final acceptance May 24, 2014. Online prepub date: June 3, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Hsin-I Ma, Department of Neurological Surgery, Tri-Service General Hospital and Graduate Institute of Medical Sciences, National Defense Medical Center, No. 325, Sec. 2, Cheng-Kung Road, Taipei 11490, Taiwan. Tel: +886-2-8792-3311; Fax: +886-2-8792-7178; E-Mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1451-1467, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683034
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Electrophysiological Characterization of eGFP-Labeled Intrastriatal Dopamine Grafts

Meltem Hohmann,*† Regina Rumpel,* Martin Fischer,‡ Malte Donert,* Andreas Ratzka,* Alexander Klein,* Maike Wesemann,* Anna Effenberg,* Christoph Fahlke,§ and Claudia Grothe*†

*Institute of Neuroanatomy, Hannover Medical School, Hannover, Germany
†Center for Systems Neuroscience, Hannover, Germany
‡Institute of Neurophysiology, Hannover Medical School, Hannover, Germany
§Institute of Complex Systems, Zelluläre Biophysik, FZ JülichJülich, Germany

Substitution of degenerated dopaminergic (DA) neurons by intrastriatally transplanted ventral mesencephalon (VM)-derived progenitor cells has been shown to improve motor functions in parkinsonian patients and animal models, whereas investigations of electrophysiological properties of the grafted DA neurons have been rarely performed. Here we show electrophysiological properties of grafted VM progenitor cells at different time intervals up to 12 weeks after transplantation measured in acute brain slices using eGFP-Flag transfection to identify the graft. We were able to classify typical DA neurons according to the biphasic progression (voltage “sag”) to hyperpolarizing current injections. Two types of DA-like neurons were classified. Whereas type 1 neurons were characterized by delayed action potentials after hyperpolarization and irregular spontaneous firing, type 2 neurons displayed burst firing after hyperpolarization, spontaneous bursts, and regular firing. Comparison to identified DA neurons in vitro indicates a high integration of the intrastriatally grafted neurons, since in vitro cultures displayed regular firing spontaneously, whereas grafted identified DA neurons showed irregular firing. Additionally, type 1 and type 2 neurons exhibited a slight increase in the spontaneous firing frequency over time intervals after grafting, which might reflect a progressive integration of the grafted DA neurons. Our results provide evidence of the differentiation of grafted VM progenitor cells into mature integrated DA neurons, which are shown to replace the missing DA neurons functionally early after grafting.

Key words: Parkinson’s disease (PD); Dopaminergic (DA) neurons; Cell replacement; Patch clamp technique; Enhanced green fluorescent protein (eGFP) transfection

Received February 14, 2014; final acceptance July 11, 2014. Online prepub date: July 15, 2014.
Address correspondence to Prof. Dr. C. Grothe, Hannover Medical School, Institute of Neuroanatomy (OE4140), Carl-Neuberg-Strasse 1, 30625 Hannover, Germany. Tel: +49 511 532-2897; Fax: +49 511 532-2880; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1469-1479, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682413
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Feasibility and Safety of Intra-arterial Pericyte Progenitor Cell Delivery Following Mannitol-Induced Transient Blood–Brain Barrier Opening in a Canine Model

Sung Won Youn,*1 Keun-Hwa Jung,†‡1 Kon Chu,†‡ Jong-Young Lee,§ Soon-Tae Lee,†‡ Jae-jun Bahn,‡ Dong-Kyu Park,‡ Jung-Suk Yu,‡ So-Yun Kim,‡ Manho Kim,†‡ Sang Kun Lee,†‡ Moon-Hee Han,¶ and Jae-Kyu Roh†‡#

*Department of Neuroradiology, Catholic University of Daegu Medical Center, School of Medicine, Catholic University of Daegu, Daegu, South Korea
†Department of Neurology, Seoul National University Hospital, College of Medicine, Seoul National University, Seoul, South Korea
‡Laboratory for Neurotherapeutics, Biomedical Research Institute, Seoul National University Hospital, Seoul, South Korea
§Department of Neurosurgery, Kangdong Sacred Heart Hospital, Seoul, South Korea
¶Department of Radiology, Seoul National University Hospital, College of Medicine, Seoul National University, Seoul, South Korea
#Department of Neurology, The Armed Forces Capital Hospital, Sungnam, South Korea

Stem cell therapy is currently being studied with a view to rescuing various neurological diseases. Such studies require not only the discovery of potent candidate cells but also the development of methods that allow optimal delivery of those candidates to the brain tissues. Given that the blood–brain barrier (BBB) precludes cells from entering the brain, the present study was designed to test whether hyperosmolar mannitol securely opens the BBB and enhances intra-arterial cell delivery. A noninjured normal canine model in which the BBB was presumed to be closed was used to evaluate the feasibility and safety of the tested protocol. Autologous adipose tissue-derived pericytes with platelet-derived growth factor receptor b positivity were utilized. Cells were administered 5 min after mannitol pretreatment using one of following techniques: (1) bolus injection of a concentrated suspension, (2) continuous infusion of a diluted suspension, or (3) bolus injection of a concentrated suspension that had been shaken by repeated syringe pumping. Animals administered a concentrated cell suspension without mannitol pretreatment served as a control group. Vital signs, blood parameters, neurologic status, and major artery patency were kept stable throughout the experiment and the 1-month posttreatment period. Although ischemic lesions were noted on magnetic resonance imaging in several mongrel dogs with concentrated cell suspension, the injection technique using repeated syringe shaking could avert this complication. The cells were detected in both ipsilateral and contralateral cortices and were more frequent at the ipsilateral and frontal locations, whereas very few cells were observed anywhere in the brain when mannitol was not preinjected. These data suggest that intra-arterial cell infusion with mannitol pretreatment is a feasible and safe therapeutic approach in stable brain diseases such as chronic stroke.

Key words: Adipose tissue-derived, platelet-derived growth factor receptor β-positive (AT-PDGFRβ+) cells; Canine; Intra-arterial injection; Hyperosmolar mannitol; Blood–brain barrier (BBB)

Received November 17, 2013; final acceptance June 12, 2014. Online prepub date: June 13, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Jae-Kyu Roh, M.D., Ph.D., Department of Neurology, Seoul National University Hospital, 28, Yongon-dong, Chongro-gu, Seoul 110-744, South Korea. Tel: +82-2-20723265; Fax: +82-2-36724949; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Kon Chu, M.D., Ph.D., Department of Neurology, Seoul National University Hospital, 28, Yongon-dong, Chongro-gu Seoul, 110-744, South Korea. Tel: +82-2-20721878; Fax: +82-2-36724949; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1481-1492, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682800
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Early Inflammatory Responses Following Cell Grafting in the CNS Trigger Activation of the Subventricular Zone: A Proposed Model of Sequential Cellular Events

Jelle Praet,*†‡ Eva Santermans,§ Jasmijn Daans,*† Debbie Le Blon,*† Chloe Hoornaert,*† Herman Goossens,† Niel Hens,†§¶ Annemie Van der Linden,‡ Zwi Berneman,*† and Peter Ponsaerts*†

*Experimental Cell Transplantation Group, Laboratory of Experimental Hematology, University of Antwerp, Wilrijk, Belgium
†Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Wilrijk, Belgium
BioImaging Laboratory, University of Antwerp, Wilrijk, Belgium
§Center for Statistics, I-Biostat, Hasselt University, Diepenbeek, Belgium
¶Centre for Health Economic Research and Modeling Infectious Diseases (Chermid), University of Antwerp, Wilrijk, Belgium

While multiple rodent preclinical studies, and to a lesser extent human clinical trials, claim the feasibility, safety, and potential clinical benefit of cell grafting in the central nervous system (CNS), currently only little convincing knowledge exists regarding the actual fate of the grafted cells and their effect on the surrounding environment (or vice versa). Our preceding studies already indicated that only a minor fraction of the initially grafted cell population survives the grafting process, while the surviving cell population becomes invaded by highly activated microglia/macrophages and surrounded by reactive astrogliosis. In the current study, we further elaborate on early cellular and inflammatory events following syngeneic grafting of eGFP
+ mouse embryonic fibroblasts (mEFs) in the CNS of immunocompetent mice. Based on obtained quantitative histological data, we here propose a detailed mathematically derived working model that sequentially comprises hypoxia-induced apoptosis of grafted mEFs, neutrophil invasion, neoangiogenesis, microglia/macrophage recruitment, astrogliosis, and eventually survival of a limited number of grafted mEFs. Simultaneously, we observed that the cellular events following mEF grafting activates the subventricular zone neural stem and progenitor cell compartment. This proposed model therefore further contributes to our understanding of cell graft-induced cellular responses and will eventually allow for successful manipulation of this intervention.

Key words: Cell transplantation; Graft apoptosis; Immune response; Angiogenesis; Cell proliferation

Received March 4, 2014; final acceptance July 8, 2014. Online prepub date: July 14, 2014.
Address correspondence to Prof. Dr. Peter Ponsaerts, Experimental Cell Transplantation Group, Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Campus Drie Eiken (CDE-S6.51), 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. 24, pp. 1493-1509, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682828
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Purinergic Receptors in Spinal Cord-Derived Ependymal Stem/Progenitor Cells and Their Potential Role in Cell-Based Therapy for Spinal Cord Injury

Rosa Gomez-Villafuertes,* Francisco Javier Rodriguez-Jimenez,† Ana Alastrue-Agudo,† Miodrag Stojkovic,‡§ Maria Teresa Miras-Portugal,*1 and Victoria Moreno-Manzano†1

*Departamento de BioquimicaFacultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain
†Neuronal and Tissue Regeneration Lab., Centro de Investigacion Principe Felipe, Valencia, Spain
Spebo Medical, Leskovac, Serbia
§Human Genetics, Faculty of Medical Sciences, University of Kragujevac, Serbia

Spinal cord injury (SCI) is a major cause of paralysis with no current therapies. Following SCI, large amounts of ATP and other nucleotides are released by the traumatized tissue leading to the activation of purinergic receptors that, in coordination with growth factors, induce lesion remodeling and repair. We found that adult mammalian ependymal spinal cord-derived stem/progenitor cells (epSPCs) are capable of responding to ATP and other nucleotidic compounds, mainly through the activation of the ionotropic P2X4, P2X7, and the metabotropic P2Y1
and P2Y4 purinergic receptors. A comparative study between epSPCs from healthy rats versus epSPCis, obtained after SCI, shows a downregulation of P2Y1 receptor together with an upregulation of P2Y4 receptor inepSPCis. Moreover, spinal cord after severe traumatic contusion shows early and persistent increases in the expression of P2X4 and P2X7 receptors around the injury, which are completely reversed when epSPCis were ectopically transplanted. Since epSPCitransplantation significantly rescues neurological function after SCI in parallel to inhibition of the induced P2 ionotropic receptors, a potential avenue is open for therapeutic alternatives in SCI treatments based on purinergic receptors and the endogenous reparative modulation.

Key words: Purinergic receptors; Pharmacological properties; Ependymal cells; Stem/progenitor cells; Spinal cord injury (SCI); Locomotion recovery

Received March 27, 2014; final acceptance July 10, 2014. Online prepub date: July 15, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Rosa Gomez-Villafuertes, Ph.D., Biochemistry Department, School of Veterinary, Universidad Complutense, Av. Puerta de Hierro s/n, 28040 Madrid, Spain. Tel: +34913943890; Fax: +34913943909; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1511-1532, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683511
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

3,2-Dihydroxyflavone-Treated Pluripotent Stem Cells Show Enhanced Proliferation, Pluripotency Marker Expression, and Neuroprotective Properties

Dawoon Han,*1 Han Jun Kim,†1 Hye Yeon Choi,* BongWoo Kim,* Gwangmo Yang,* Jihae Han,* Ahmed Abdal Dayem,* Hye-Rim Lee,† Jin Hoi Kim,* Kyung-Mi Lee,‡ Kyu-Shik Jeong,§ Sun Hee Do,† and Ssang-Goo Cho*

*Department of Animal Biotechnology, Animal Resources Research Center, and Incurable Disease Animal Model and Stem Cell Institute (IDASI), Konkuk University, Gwangjin-Gu, Seoul, Republic of Korea
†Department of Veterinary Clinical Pathology, College of Veterinary Medicine, Konkuk University, Gwangjin-Gu, Seoul, Republic of Korea
‡Global Research Laboratory, Department of Biochemistry and Molecular Biology, Korea University College of Medicine, Seoul, Republic of Korea
§College of Veterinary Medicine, Kyungpook National University, Daegu City, Republic of Korea

Efficient maintenance of the undifferentiated status of embryonic stem cells (ESCs) may be important for preparation of high-quality cell sources that can be successfully used for stem cell research and therapy. Here we tried to identify a compound that can enhance the quality of pluripotent stem cells. Treatment of ESCs and induced pluripotent stem cells (iPSCs) with 3,2′-dihydroxyflavone (3,2′-DHF) led to increases in cell growth, colony formation, and cell proliferation. Treatment with 3,2′-DHF resulted in high expression of pluripotency markers (OCT4, SOX2, and NANOG) and significant activation (STAT3 and AKT) or suppression (GSK3β and ERK) of self-renewal-related kinases. 3,2′-DHF-treated high-quality pluripotent stem cells also showed enhanced differentiation potential. In particular, treatment of iPSCs with 3,2′-DHF led to elevated expression of ectodermal differentiation markers and improved differentiation into fully matured neurons. Next, we investigated the in vivo effect of 3,2′-DHF-pretreatediPSCs (3,2′-DHF iPSCs) in a peripheral nerve injury model and found that transplantation of 3,2′-DHF iPSCs resulted in more efficient axonal regeneration and functional recovery than in controls. Upon histopathological and gene expression analyses, we found that transplantation of 3,2′-DHF iPSCs stimulated expression of cytokines, such as TNF-a, in the early phase of injury and successfully reduced convalescence time of the injured peripheral nerve, showing an effective neuroprotective property. Taken together, our data suggest that 3,2′-DHF can be used for more efficient maintenance of pluripotent stem cells as well as for further applications in stem cell research and therapy.

Key words: 3,2′-Dihydroxyflavone; Pluripotent stem cell; Self-renewal; Peripheral nerve injury model; Neuroprotective property

Received November 1, 2013; final acceptance July 26, 2014. Online prepub date: July 30, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Ssang-Goo Cho, Department of Animal Biotechnology and Animal Resources Research Center, Konkuk University, Gwangjin-GuAchasan-rho 263, Seoul 143-701, Republic of Korea. Tel: +82-2-450-4207; Fax: +82-2-450-1044; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Sun Hee Do, Department of Veterinary Clinical Pathology, College of Veterinary Medicine, Konkuk University, Gwangjin-GuAchasan-rho 263, Seoul, 143-701, Republic of Korea. Tel: +82-2-450-3706; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1533-1544, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679345
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Combined Bone Mesenchymal Stem Cell and Olfactory Ensheathing Cell Transplantation Promotes Neural Repair Associated With CNTF Expression in Traumatic Brain-Injured Rats

Xue-Mei Fu,* Su-Juan Liu,† Qi-Qin Dan,‡ Yan-Ping Wang,§ Na Lin,§ Long-Yun LV,§ Yu Zou,‡ Su Liu,§ Xue Zhou,†1 and Ting-Hua Wang†‡§1

*Shenzhen Children’s Hospital, Shenzhen, Guangdong, China
†Department of Histology and Neurobiology, West China School of Preclinical and Forensic Medicine, Chengdu, Sichuan, China
‡Institute of Neurological Disease, State Key Lab of Biotherapy, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, Sichuan, China
§Institute of Neuroscience, Kunming Medical University, Kunming, Yunnan, China

This study examined the role of bone mesenchymal stem cell (BMSC) and olfactory ensheathing cell (OEC) cografting on neural function and underlying molecular mechanisms in acute stage of traumatic brain injury (TBI) rats. Eighty Sprague–Dawley (SD) female rats were randomly divided into five groups (n = 16 per category): sham operated group (Sham), weight-drop-induced TBI group (TBI), BMSC transplantation group (BMSC), OEC transplantation group (OEC), and cotransplantation group (CO). Eight rats were randomly selected from each group for behavioral and morphological assessment. Another category (n = 8 rats) was employed in the genetic expression detection. BMSCs were isolated from GFP mice and identified by CD44 antibody. OECs were isolated from the SD rats, identified by P75 antibody and labeled by Hoechst 33342. They were then transplanted into the surrounding tissue of the epicenter of TBI rats. The result of neurological severity scores revealed that BMSC or OEC transplantation alone and BMSC and OEC cografting significantly ameliorated the neurological deficits of TBI rats. Quantitative immunohistochemical analysis showed that graft-recipient animals possessed dramatically more neurons and regenerated axons and smaller amounts of astrocytes than controls 14 days posttransplantation (p < 0.05). However, the expressional level of ciliary neurotrophic factor significantly decreased in the cografting group as determined by RT-PCR (p < 0.05), and the Janus kinase/signal transducer and activator of transcription pathway was significantly activated at 7 days after cell transplantation (p < 0.05). This study is the first to report the role of cotransplantation of BMSCs and OECs in the therapy of TBI and explore its potential molecular mechanisms, therefore providing the important morphological and molecular biological evidence for the clinical application of BMSC and/or OEC transplantation in TBI.

Key words: Bone mesenchymal stem cells (BMSCs); Olfactory ensheathing cells (OECs); Transplantation; Traumatic brain injury (TBI)

Received April 30, 2013; final acceptance February 18, 2014. Online prepub date: March 7, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Professor Ting-Hua Wang, Institute of Neuroscience, Kunming Medical University, Kunming, Yunnan 650031, China. Tel/Fax: +86-871-65922818; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1545-1554, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683043
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Expired and Pathogen-Inactivated Platelet Concentrates Support Differentiation and Immunomodulation of Mesenchymal Stromal Cells in Culture

Sandra Mjoll Jonsdottir-Buch,*† Hildur Sigurgrimsdottir,*† Ramona Lieder,* and Olafur Eysteinn Sigurjonsson*†‡

*REModeL Lab, The Blood Bank, Landspitali University Hospital, Reykjavik, Iceland
†Faculty of Medicine, Department of Biomedical Sciences, University of Iceland, Reykjavík, Iceland
‡School of Science and Engineering, Reykjavik University, Reykjavik, Iceland

Platelet lysates have been reported as suitable cell culture supplement for cultures of mesenchymal stromal cells (MSCs). The demand for safe and animal-free cultures of MSCs is linked to the potential application of MSCs in clinics. While the use of platelet lysates offers an alternative to animal serum in MSC cultures, obtaining supplies of fresh platelet concentrates for lysate production is challenging and raises concerns due to the already existing shortage of platelet donors. We have previously demonstrated that expired platelet concentrates may represent a good source of platelets for lysate production without competing with blood banks for platelet donors. The INTERCEPT Blood System™ treatment of platelet concentrates allows for prolonged storage up to 7 days, using highly specific technology based on amotosalen and UV-A light. The INTERCEPT system has therefore been implemented in blood processing facilities worldwide. In this study, we evaluated the suitability of INTERCEPT-treated, expired platelet concentrates, processed into platelet lysates, for the culture of MSCs compared to nontreated expired platelets. Bone marrow-derived MSCs were cultured in media supplemented with either platelet lysates from traditionally prepared expired platelet concentrates or in platelet lysates from expired and pathogen-inactivated platelet concentrates. The effects of pathogen inactivation on the ability of the platelets to support MSCs in culture were determined by evaluating MSC immunomodulation,immunophenotype, proliferation, and trilineage differentiation. Platelet lysates prepared from expired and pathogen-inactivated platelet concentrates supported MSC differentiation and immunosuppression better compared to traditionally prepared platelet lysates from expired platelet units. Pathogen inactivation of platelets with the INTERCEPT system prior to use in MSC culture had no negative effects on MSC immunophenotype or proliferation. In conclusion, the use of expired pathogen-inactivated platelet units from blood banks to prepare platelet lysates for the culture of MSCs is desirable and attainable.

Key words: Mesenchymal stromal cells (MSCs); Differentiation; Pathogen inactivation; Platelet; Platelet lysate; Amotosalen; INTERCEPT

Received December 10, 2013; final acceptance July 11, 2014. Online prepub date: July 15, 2014.
Address correspondence to Olafur E. SigurjonssonREModeL Lab, The Blood Bank, Landspitali University Hospital, Snorrabraut 60, 105, Reykjavik, Iceland. Tel: +3545435523; Fax: +3545435532; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1555-1570, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683016
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Undifferentiated Wharton’s Jelly Mesenchymal Stem Cell Transplantation Induces Insulin-Producing Cell Differentiation and Suppression of T-Cell-Mediated Autoimmunity in Nonobese Diabetic Mice

Pei-Jiun Tsai,*† Hwai-Shi Wang,‡ Gu-Jiun Lin,§ Shu-Cheng Chou,¶ Tzu-Hui Chu,§ Wen-Ting Chuan,§ Ying-Jui Lu,§ Zen-Chung Weng,#** Cheng-Hsi Su,†† Po-Shiuan Hsieh,‡‡ Huey-Kang Sytwu,§§ Chi-Hung Lin,*¶¶ Tien-Hua Chen,‡¶1 and Jia-Fwu Shyu§1

*Institute of Clinical Medicine, National Yang Ming University, Taipei, Taiwan
†Department of Critical Care Medicine, Veteran General Hospital, Taipei, Taiwan
‡Institute of Anatomy and Cell Biology, School of Medicine, National Yang Ming University, Taipei, Taiwan
§Department of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan
¶Department of Surgery, Veteran General Hospital, Taipei, Taiwan
#Division of Cardiovascular Surgery, Department of Surgery, Taipei Medical University Hospital, Taipei, Taiwan
**Department of Surgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan
††Department of Surgery, Cheng Hsin General Hospital, Taipei, Taiwan
‡‡Department of Physiology and Biophysics, National Defense Medical Center, Taipei, Taiwan
§§Department of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan
¶¶Institute of Microbiology and Immunology, National Yang Ming University, Taipei, Taiwan

Type 1 diabetes mellitus is caused by T-cell-mediated autoimmune destruction of pancreatic β-cells. Systemic administration of mesenchymal stem cells (MSCs) brings about their incorporation into a variety of tissues with immunosuppressive effects, resulting in regeneration of pancreatic islets. We previously showed that human MSCs isolated from Wharton’s jelly (WJ-MSCs) represent a potential cell source to treat diabetes. However, the underlying mechanisms are unclear. The purpose of this study was to discern whether undifferentiated WJ-MSCs can differentiate into pancreatic insulin-producing cells (IPCs) and modify immunological responses in nonobese diabetic (NOD) mice. Undifferentiated WJ-MSCs underwent lentiviral transduction to express green fluorescent protein (GFP) and then were injected into the retro-orbital venous sinus of NOD mice. Seven days after transplantation, fluorescent islet-like cell clusters in the pancreas were apparent. WJ-MSC-GFP-treated NOD mice had significantly lower blood glucose and higher survival rates than saline-treated mice. Systemic and local levels of autoaggressive T-cells, including T helper 1 cells and IL-17-producing T-cells, were reduced, and regulatory T-cell levels were increased. Furthermore, anti-inflammatory cytokine levels were increased, and dendritic cells were decreased. At 23 days, higher human C-peptide and serum insulin levels and improved glucose tolerance were found. Additionally, WJ-MSCs-GFP differentiated into IPCs as shown bycolocalization of human C-peptide and GFP in the pancreas. Significantly more intact islets and less severe insulitis were observed. In conclusion, undifferentiated WJ-MSCs can differentiate into IPCs in vivo with immunomodulatory effects and repair the destroyed islets in NOD mice.

Key words: Wharton’s jelly; Mesenchymal stem cells (MSCs); Insulin-producing cells (IPCs); Immunomodulation; NOD mice

Received November 17, 2013; final acceptance July 10, 2014. Online prepub date: July 15, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Jia-Fwu Shyu, M.D., Ph.D., Department of Biology and Anatomy, National Defense Medical Center, 161 Ming Chuan E. Road Section 6, Taipei, Taiwan 114, R.O.C. Tel: +(886)-2-87922484; Fax: +(886)-2-87922484; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Dr. Tien-Hua Chen, M.D., Institute of Anatomy and Cell Biology, School of Medicine, National Yang Ming University, 201 Shih-Pai Road Section 2, Taipei, Taiwan 112, R.O.C. Tel: +(886)-2-2-28267035; Fax: +(886)-2-28212884; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1571-1584, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682792
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Diabetic Mesenchymal Stem Cells Are Ineffective for Improving Limb Ischemia Due to Their Impaired Angiogenic Capability

Hyongbum Kim,*†1 Ji Woong Han,*1 Ji Yoon Lee,*‡ Yong Jin Choi,*§ Young-Doug Sohn,* Myungjae Song,† and Young-sup Yoon*

*Department of Medicine, Division of Cardiology, Emory University School of Medicine, Atlanta, GA, USA
†Graduate School of Biomedical Science and Engineering/College of Medicine, Hanyang University, Seoul, Korea
‡Department of Hematology, College of Medicine, The Catholic University of Korea, Seoul, Korea
§Division of Cellular and Developmental Biology, Molecular and Cell Biology Department, University of California at Berkeley, Berkeley, CA, USA

The purpose of this study was to investigate the effects of diabetes on mesenchymal stem cells (MSCs) in terms of their angiogenic and therapeutic potential for repairing tissue ischemia. We culture-isolated MSCs from streptozotocin-induced diabetic rats (D-MSCs) and compared their proliferation, differentiation, and angiogenic effects with those from normal rats (N-MSCs). The angiogenic effects of MSCs were evaluated by real-time PCR, in vitro tube formation assay, and transplantation of the MSCs into ahindlimb ischemia model followed by laser Doppler perfusion imaging. The number of MSCs derived from diabetic rats was smaller, and their proliferation rate was slower than N-MSCs. Upon induction of differentiation, the osteogenic and angiogenicdifferentiation of D-MSCs were aberrant compared to N-MSCs. The expression of angiogenic factors was lower in D-MSCs than N-MSCs. D-MSCs cocultured with endothelial cells resulted in decreased tube formation compared to N-MSCs. D-MSCs were ineffective to improve hindlimb ischemia and showed lower capillary density and angiogenic gene expression in ischemic limbs than N-MSCs. D-MSCs have defective proliferation and angiogenic activities and are ineffective for repairing hindlimb ischemia. Newer measures are needed before MSCs can be employed as a source for autologous cell therapy.

Key words: Diabetes mellitus; Mesenchymal stem cells (MSCs); Ischemic; Angiogenesis

Received January 26, 2012; final acceptance June 30, 2014. Online prepub date: July 08, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Young-sup Yoon, M.D., Ph.D., Department of Medicine, Division of Cardiology, Emory University, 101 Woodruff Circle, WMB 3309, Atlanta, GA 30322, USA. Tel: +1-404-727-8176; Fax: +1-404-727-3988; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1584-1598, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682819
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Clinical Immunosuppressants Inhibit Inflammatory, Proliferative, and Reprogramming Potential, But Not Angiogenesis of Human Pancreatic Duct Cells

Lei Ding,*1 Yves Heremans,†1 Daniel Pipeleers,‡ Zhidong Ling,‡ Harry Heimberg,† Conny Gysemans,*2 and Chantal Mathieu*2

*Laboratory of Clinical and Experimental Endocrinology, Campus Gasthuisberg O&N1, Faculty of Medicine, Katholieke Universiteit Leuven (KU Leuven), Leuven, Belgium
†Laboratory of Beta Cell Neogenesis (BENE), Diabetes Research Center, Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel (VUB), Brussels, Belgium
‡Laboratory of Diabetes Pathology and Therapy, Diabetes Research Center, Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel (VUB), Brussels, Belgium

The presence of pancreatic duct cells in clinical islet grafts may affect long-term metabolic success. Human pancreatic duct cells express factors that may exert both protective and damaging effects on islet cells in the graft. Here we studied the potential of commonly used immunosuppressive drugs in islet transplantation—sirolimus, tacrolimus, and mycophenolate mofetil (MMF)—to influence the inflammatory and angiogenic capacity of human pancreatic duct cells in addition to their proliferation and reprogramming abilities. Our data show that the expression of specific proinflammatory cytokines by the human pancreatic duct cells was either unaltered or inhibited by the immunosuppressants studied, especially tacrolimus and MMF, whereas expression of chemotactic and angiogenic factors was unaffected. Although none of the immunosuppressants directly led to duct cell death, MMF prevented duct cell proliferation, and sirolimus inhibited neurogenin 3-mediated duct-to-(neuro)endocrine cell reprogramming. Our data indicate that the immunosuppressant tacrolimus was the least aggressive on the angiogenic, proliferative, and reprogramming potential of human pancreatic duct cells, while it was most powerful in inhibiting inflammatory cytokines, which may influence the outcome of islet transplantation.

Key words: Human pancreatic duct cell; Islet transplantation; Immunosuppressant; Cytokines; Angiogenesis; Dedifferentiation

Received November 28, 2013; final acceptance July 9, 2014. Online prepub date: July 14, 2014.
1These authors provided equal contribution to this work.
2Share senior authorship.
Address correspondence to Chantal Mathieu, M.D., Ph.D., Campus Gasthuisberg O&N1, Laboratory of Clinical and Experimental Endocrinology, Herestraat 49 bus 902, B-3000 Leuven, Belgium. Tel: +32-16-346023; Fax: +32-16-330718; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1599-1614, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X678571
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

TIM4 Regulates the Anti-Islet Th2 Alloimmune Response

Andrea Vergani,*†1 Francesca Gatti,*‡1 Kang M. Lee Francesca D’Addio,*† Sara Tezza,* Melissa Chin,* Roberto Bassi,* Ze Tian,¶ Erxi Wu,# Paola Maffi,† Moufida Ben Nasr,* James I. Kim,§ AntonioSecchi,†** James F. Markmann,§ David M. Rothstein,†† Laurence A. Turka,‡‡ Mohamed H. Sayegh,* and Paolo Fiorina*†

*Transplantation Research Center, Division of Nephrology, Boston Children’s Hospital and Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA
†Transplant Medicine, Ospedale San Raffaele, Milan, Italy
‡University of Salento, Lecce, Italy
§Transplant Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
¶Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA
#Department of Pharmaceutical Sciences, North Dakota State University, Fargo, ND, USA
**Vita-Salute San Raffaele University, Milan, Italy
††Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA
‡‡Department of Surgery, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

The role of the novel costimulatory molecule TIM4 in anti-islet response is unknown. We explored TIM4 expression and targeting in Th1 (BALB/c islets into C57BL/6 mice) and Th2 (BALB/c islets into Tbet−/−
C57BL/6 mice) models of anti-islet alloimmuneresponse and in a model of anti-islet autoimmune response (diabetes onset in NOD mice). The targeting of TIM4, using the monoclonal antibody RMT4-53, promotes islet graft survival in a Th1 model, with 30% of the graft surviving in the long term; islet graft protection appears to be mediated by a Th1 to Th2 skewing of the immune response. Differently, in the Th2 model, TIM4 targeting precipitates graft rejection by further enhancing the Th2 response. The effect of anti-TIM4 treatment in preventing autoimmune diabetes was marginal with only minor Th1 to Th2 skewing. B-Cell depletion abolished the effect of TIM4 targeting. TIM4 is expressed on human B-cells and is upregulated in diabetic and islet-transplanted patients. Our data suggest a model in which TIM4 targeting promotes Th2 response over Th1 via B-cells. The targeting of TIM4 could become a component of an immunoregulatory protocol in clinical islet transplantation, aiming at redirecting the immune system toward a Th2 response.

Key words: Islet transplantation; Autoimmune diabetes; Costimulatory molecules; Regulatory cells

Received March 3, 2013; final acceptance January 15, 2014. Online prepub date: March 7, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Paolo Fiorina, M.D. Ph.D., Nephrology Division, Enders Building 5
th Floor Room EN530, Boston Children’s Hospital, Harvard Medical School, Boston, 02115 MA, USA. Tel: +1 617-919-2624; Fax: +1 617-732-5254; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1615-1626, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X672440
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Improved Insulin Secretion by Autologous Islet Transplantation, Compared to Oral Antidiabetic Agents, After Distal Pancreatectomy

Ji Won Yoon,*† Hye Seung Jung,*‡ Jin Young Jang,§ Min Joo Kim,*1 Jung Hee Kim,* Jung Hun Ohn,* Jae Hyeon Kim,*2 Hak Mo Lee,‡ Hyo Cheol Kim,¶ Kyoung Bun Lee,# Seung A Choi,† Sun-WheKim,§ and Kyong Soo Park*‡

*Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea
†Healthcare Research Institute, Seoul National University Hospital Healthcare System Gangnam Center, Seoul, Korea
‡Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea
§Department of Surgery, Seoul National University College of Medicine, Seoul, Korea
¶Department of Radiology, Seoul National University College of Medicine, Seoul, Korea
#Department of Pathology, Seoul National University College of Medicine, Seoul, Korea

In this study, the effects of autologous islet transplantation (ITx) were compared to those of oral antidiabetic drugs (OAD) after distal pancreatectomy (NCT01922492). We enrolled nondiabetic patients who underwent distal pancreatectomy for benign tumors. In the ITx group, islets were isolated from the normal part of the resected pancreas and implanted via the portal vein. Patients who did not receive ITx were regularly monitored and were enrolled in the OAD group if diabetes mellitus developed. The OAD group was treated with metformin with or without vildagliptin. Metabolic parameters were monitored for 12 months postoperatively. Nine patients in the ITx group and 10 in the OAD group were included in the analysis. After 12 months, hemoglobin A1csignificantly increased by 5% of the baseline in each group. Area under the curve for blood glucose (AUCglucose) of the 75-g oral glucose tolerance test increased similarly in the immediate postoperative period in both groups but significantly reduced only in the ITx group thereafter. Insulinogenic index (INSindex) significantly decreased from 25.6 ± 18.9 to 4.7 ± 3.7 in the OAD group, while no significant change was observed in the ITx group (from 15.0 ± 4.5 to 11.0 ± 8.2). In the multiple regression analysis,ITx was an independent factor for changes in AUCglucose
and INSindex. In addition, changes in INSindex in the ITx group after postoperative 6 months were associated with the efficacy of islet isolation, amount of grafts, and peak serum HMGB1 and VEGF levels after ITxITx was superior to OAD in maintaining insulin secretory capacity and glucose tolerance after distal pancreatectomy.

Key words: Autologous islet transplantation (ITx); Partial pancreatectomy; Glucose tolerance; Glucose-stimulated insulin secretion; Pancreatogenic diabetes

Received December 27, 2013; final acceptance June 25, 2014. Online prepub date: June 27, 2014.
1Current address: Department of Internal Medicine, Korea Cancer Center Hospital, Seoul, Korea.
2Current address: Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. Address correspondence to Hye Seung Jung, M.D., Ph.D., Department of Internal Medicine, Seoul National University College of Medicine, 101Daehak-roJongno-gu, Seoul 110-744, Republic of Korea. Tel: +82-2-2072-0240; Fax: +82-2-762-9662; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Kyong Soo Park, M.D., Ph.D., Department of Internal Medicine, Seoul National University College of Medicine, 101 Daehak-roJongno-gu, Seoul 110-744, Republic of Korea. Tel: +82-2-2072-2946; Fax: +82-2-762-9662; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1627-1638, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682422
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Phases I–II Matched Case-Control Study of Human Fetal Liver Cell Transplantation for Treatment of Chronic Liver Disease

Giada Pietrosi,* Giovanni Vizzini,* Jorg Gerlach,† Cinzia Chinnici,‡ Angelo Luca,§ Giandomenico Amico,‡ Monica D’Amato,‡ Pier Giulio Conaldi,¶ Sergio Li Petri,# Marco Spada,# Fabio Tuzzolino,** Luigi Alio,†† Eva Schmelzer,† and Bruno Gridelli#

*Hepatology Unit, Department of Medicine, Mediterranean Institute for Transplantation and Advanced Specialized Therapies (IRCCS-ISMETT), Palermo, Italy
†McGowan Institute for Regenerative Medicine, Departments of Surgery and Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA
‡Fondazione Ri.MED, Palermo, Italy
§Department of Diagnostic and Therapeutics Services, Mediterranean Institute for Transplantation and Advanced Specialized Therapies (IRCCS-ISMETT), Palermo, Italy
¶Department of Laboratory Medicine and Advanced Biotechnologies, Mediterranean Institute for Transplantation and Advanced Specialized Therapies (IRCCS-ISMETT), Palermo, Italy
#Abdominal and Transplantation Surgery Unit, Department of Surgery, Mediterranean Institute for Transplantation and Advanced Specialized Therapies (IRCCS-ISMETT), Palermo, Italy
**Research Office, Mediterranean Institute for Transplantation and Advanced Specialized Therapies (IRCCS-ISMETT), Palermo, Italy
††Department of Obstetrics and Gynecology, Civico Hospital, Palermo, Italy

Fetal hepatocytes have a high regenerative capacity. The aim of the study was to assess treatment safety and clinical efficacy of human fetal liver cell transplantation through splenic artery infusion. Patients with endstage chronic liver disease on the waiting list for liver transplantation were enrolled. A retrospectively selected contemporary matched-pair group served as control. Nonsorted raw fetal liver cell preparations were isolated from therapeutically aborted fetuses. The end points of the study were safety and improvement of the Model for End-Stage Liver Disease (MELD) and Child-Pugh scores. Nine patients received a total of 13 intrasplenic infusions and were compared with 16 patients on standard therapy. There were no side effects related to the infusion procedure. At the end of follow-up, the MELD score (mean ± SD) in the treatment group remained stable from baseline (16.0 ± 2.9) to the last observation (15.7 ± 3.8), while it increased in the control group from 15.3 ± 2.5 to 19 ± 5.7 (p = 0.0437). The Child-Pugh score (mean ± SD) dropped from 10.1 ± 1.5 to 9.1 ± 1.4 in the treatment group and increased from 10.0 ± 1.2 to 11.1 ± 1.6 in the control group (p = 0.0076). All treated patients with history of recurrent portosystemic encephalopathy (PSE) had no further episodes during 1-year follow-up. No improvement was observed in the control group patients with PSE at study inclusion. Treatment was considered a failure in six of the nine patients (three deaths not liver related, one liver transplant, two MELD score increases) compared with 14 of the 16 patients in the control group (six deaths, five of which were caused by liver failure, four liver transplants, and four MELD score increases). Intrasplenic fetal liver cell infusion is a safe and well-tolerated procedure in patients with end-stage chronic liver disease. A positive effect on clinical scores and on encephalopathy emerged from this preliminary study.

Key words: Human fetal liver cell transplantation (hFLCTx); Fetal hepatocytes; Chronic liver disease; Model for End-Stage Liver Disease (MELD) score; Portosystemic encephalopathy (PSE)

Received July 7, 2013; final acceptance June 20, 2014. Online prepub date: June 26, 2014.
Address correspondence to Giada Pietrosi, M.D., IRCCS-ISMETT, Via E. Tricomi 5, 90127, Palermo, Italy. Tel: +0039 335 786 54 48; Fax: +0039 091 21 92 288; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1639-1652, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681955
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Progenitor Cell Quantification After Xenotransplantation in Rat and Mouse Models by a Sensitive qPCR Assay

Julie Prigent,* Astrid Herrero,* Jérôme Ambroise,† Françoise Smets,* Gisèle A. Deblandre,*1 and Etienne M. Sokal*1

*Université Catholique de Louvain, Institut de Recherche Expérimentale et Clinique, Paediatric Hepatology and Cell Therapy, Brussels, Belgium
Université Catholique de Louvain, Institut de Recherche Expérimentale et Clinique, Center for Applied Molecular Technologies (CTMA), Brussels, Belgium

Xenotransplantation of human cells in animal models is an essential tool for evaluation of safety and efficacy of cell-based products for therapeutic use. Sensitive and reproducible methods are needed to detect and quantify human cells engrafted into the host tissue either in the targeted organ or in undesired locations. We developed a robust quantitative polymerase chain reaction (qPCR) assay based on amplification of human AluYb8 repeats, to assess the number of human cells present in rat or mouse tissues after transplantation. Standard curves of mixed human/rodent DNA and mixed human/rodent cells have been performed to determine the limit of detection and linear range of the assay. Standard curves from DNA mixing differed significantly from standard curves from cell mixing. We show here that the AluYb8 qPCR assay is highly reproducible and is able to quantify human cells in a rodent cell matrix over a large linear range that extends from 50% to 0.01% human cells. Short-term in vivo studies showed that human cells could be quantified in mouse liver up to 7 days after intrasplenic transplantation and in rat liver 4 h after intrahepatic transplantation.

Key words: Hepatic progenitor cells; Xenotransplantation; Quantitative polymerase chain reaction (qPCR); Alu repeat sequences; Engraftment analysis

Received May 20, 2013; final acceptance May 17, 2014. Online prepub date: May 20, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Professor Etienne M. SokalCliniques Universitaires Saint-Luc, Paediatric Gastroenterology and Hepatology unit, Avenue Hippocrate 10, B-1200 Brussels, Belgium. Tel: +32-27641387; Fax: +32-2-7648909; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1653-1659, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X682783
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Subthreshold High-Frequency Electrical Field Stimulation Induces VEGF Expression in Cardiomyocytes

Gediminas Rackauskas,*†1 Erol Saygili,‡1 Obaida R. Rana,‡ Esra Saygili,§ Christopher Gemein,* Aleksandras Laucevicius,† Audrius Aidietis,† Germanas Marinskis,† Pranas Serpytis,† Jurgita PlisieneDainius H. Pauza,# and Patrick Schauerte**

*Department of Cardiology, University Hospital, Aachen, Germany
†Department of Cardiovascular Medicine, Vilnius University Hospital, Santariskiu Klinikos, Vilnius University, Vilnius, Lithuania
‡Division of Cardiology, Pulmonology, and Vascular Medicine, Medical Faculty, University Hospital, Düsseldorf, Germany
§Clinic for Gastroenterology, Hepatology and Infectious Diseases, Medical Faculty, University Hospital, Düsseldorf, Germany
¶Hospital of Lithuanian University of Health Sciences Kauno Klinikos, Department of Cardiology, Kaunas, Lithuania
#Institute of Anatomy, Lithuanian Health Science University, Kaunas, Lithuania
**Cardiology Center Berlin, Germany

Subthreshold electrical stimulation (SES) has been shown to induce an improvement of angiogenesis in ischemic and nonischemic skeletal muscles, mediated by increased VEGF expression. VEGF plays a key role in physiological and pathological angiogenesis. Cardiomyocytes possess the ability to synthesize and secrete VEGF. Thus, we thought to investigate the effect of SES on VEGF regulation in cultured neonatal rat ventricular myocytes (NRVMs), in the aim to reveal new techniques for therapeutic angiogenesis in ischemic heart disease. Cell cultures of NRVMs were electrically stimulated with field strengths below the myocyte depolarization threshold (0.5 V/cm with 1 ms bipolar impulse duration). Frequencies ranging from 5 Hz up to 25, 50, and 99 Hz were applied over a period of 48 h. The expression of VEGF and its receptor KDR was determined with Western blot and ELISA. To reveal the biological activity of the secreted VEGF amount, cultured human coronary artery endothelial cells (HCAECs) were treated with the cell culture supernatant of NRVMs exposed to SES. A dominant effect of SES was observed at 25 Hz. Within this particular frequency the VEGF protein amount in the cytoplasm as well as in the cell culture supernatant increased significantly. In parallel, the protein expression of the KDR receptor decreased in a significant manner. Moreover, cell culture supernatant of NRVMs exposed to SES augmented the growth of HCAECs. Cardiomyocytes respond to SES with an increase in biologically active VEGF expression that promotes cell proliferation of HCAECs. This mechanism may provide new approaches to develop therapeutic angiogenesis in the ischemic heart.

Key words: Vascular endothelial growth factor (VEGF); Angiogenesis; Cardiomyocytes; Electrical stimulation

Received December 19, 2012; final acceptance June 27, 2014. Online prepub date: July 8, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Gediminas Rackauskas, M.D., Department of Cardiovascular Medicine, Vilnius University Hospital, Santariskiu KlinikosSantariškių g. 2, 08661 Vilnius, Lithuania. Tel: +00370-5-236 5000; Fax: +00370-5-236 5111; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it