Cell Transplantation 24(5) Abstracts

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Cell Transplantation, Vol. 24, pp. 779-796, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676925
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Inhibition of Neurosphere Formation in Neural Stem/Progenitor Cells by Acrylamide

Jong-Hang Chen,* Don-Ching Lee,* Mei-Shu Chen,* Ying-Chin Ko,† and Ing-Ming Chiu*‡

*Institute of Cellular and System Medicine, National Health Research Institutes, Miaoli, Taiwan
†Environment-Omics-Disease Research Centre, China Medical University Hospital, Taichung, Taiwan
‡Department of Internal Medicine, The Ohio State University, Columbus, OH, USA

Previous studies showed that transplantation of cultured neural stem/progenitor cells (NSPCs) could improve functional recovery for various neurological diseases. This study aims to develop a stem cell-based model for predictive toxicology of development in the neurological system after acrylamide exposure. Treatment of mouse (KT98/F1B-GFP) and human (U-1240 MG/F1B-GFP) NSPCs with 0.5 mM acrylamide resulted in the inhibition of neurosphere formation (definition of self-renewal ability in NSPCs), but not inhibition of cell proliferation. Apoptosis and differentiation of KT98 (a precursor of KT98/F1B-GFP) and KT98/F1B-GFP are not observed in acrylamide-treated neurospheres. Analysis of secondary neurosphere formation and differentiation of neurons and glia illustrated that acrylamide-treated KT98 and KT98/F1B-GFP neurospheres retain the NSPC properties, such as self-renewal and differentiation capacity. Correlation of acrylamide-inhibited neurosphere formation with cell–cell adhesion was observed in mouse NSPCs by live cell image analysis and the presence of acrylamide. Protein expression levels of cell adhesion molecules [neural cell adhesion molecule (NCAM) and N-cadherin] and extracellular signal-regulated kinases (ERK) in acrylamide-treated KT98/F1B-GFP and U-1240 MG/F1B-GFP neurospheres demonstrated that NCAM decreased and phospho-ERK (pERK) increased, whereas expression of N-cadherin remained unchanged. Analysis of AKT (protein kinase B, PKB)/b-catenin pathway showed decrease in phospho-AKT (p-AKT) and cyclin D1 expression in acrylamide-treated neurospheres of KT98/F1B-GFP. Furthermore, PD98059, an ERK phosphorylation inhibitor, attenuated acrylamide-induced ERK phosphorylation, indicating that pERK contributed to the cell proliferation, but not in neurosphere formation in mouse NSPCs. Coimmunoprecipitation results of KT98/F1B-GFP cell lysates showed that the complex of NCAM and fibroblast growth factor receptor 1 (FGFR1) is present in the neurosphere, and the amount of this complex decreases after acrylamide treatment. Our results reveal that acrylamide inhibits neurosphere formation through the disruption of the neurosphere architecture in NSPCs. The downregulation of cell–cell adhesion resulted from decreasing the levels of NCAM as well as the formation of NCAM/FGFR complex.

Key words: Neural stem/progenitor cells (NSPCs); Neurosphere formation; Acrylamide; Neural cell adhesion molecule (NCAM)

Received October 11, 2013; final acceptance December 20, 2013. Online prepub date: December 30, 2013.
Address correspondence to Ing-Ming Chiu, Institute of Cellular and System Medicine, National Health Research Institutes, 35, Keyan Rd, Miaoli 350, Taiwan. Tel: +886-37-246166; Fax: +886-2-2363-0495; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 797-809, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679228
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Improved Stem Cell-Derived Motoneuron Survival, Migration, Sprouting, and Innervation With Enhanced Expression of Polysialic Acid

Abderrahman El MaaroufDamali Moyo-Lee Yaw, and Urs Rutishauser

Cellular and Developmental Neuroscience, Department of Cell Biology, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

Motoneurons (MNs) derived from mouse embryonic stem cells (ESCs) begin to express low levels of polysialic acid (PSA) at the time when they acquire an ability to migrate and extend neurites. PSA is known to promote cell migration and process outgrowth/guidance in the developing nervous system. To test if experimentally enhanced expression of PSA would augment these cellular events, the PSA-synthesizing polysialyltransferase was introduced into ESCs. In culture, the resulting higher PSA expression specifically increased neurite outgrowth and cell migration from differentiated embryoid bodies. In addition, the MN population obtained after sorting for HB9::GFP expression showed enhanced survival as well as extensive neurite outgrowth. Following transplantation of ESC-derived MNs into an adult sciatic nerve devoid of endogenous axons, the PSA augmentation increased the numbers of axons growing toward the denervated muscles. Migration of some transplanted cells inside the nerve toward muscle was also enhanced. Moreover, higher PSA expression selectively affected target innervation. It produced greater numbers of neuromuscular junctions in a predominantly fast twitch muscle and had no effect in a slow twitch muscle. These findings suggest that engineering of PSA expression in ESC could serve as an enhancement for MN cell therapy.

Key words: Stem cells; Polysialylation; Cell engineering; Motor neurons; Transplantation; Innervation

Received August 6, 2013; final acceptance January 29, 2014. Online prepub date: March 3, 2014.
Address correspondence to Abderrahman El Maarouf, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA. Tel: +1-212-639-8343; Fax: +1-212-794-6236; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Urs Rutishauser, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA. Tel: +1-212-639-8342; Fax: +1-212-794-6236; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 811-817, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676222
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Donor-Specific Anti-HLA Antibodies in Huntington’s Disease Recipients of Human Fetal Striatal Grafts

Berardino Porfirio,*† Marco Paganini,* Benedetta Mazzanti,* Silvia Bagnoli,‡ Sandra Bucciantini,* Elena Ghelli,* Benedetta Nacmias,*‡ Anna Laura Putignano,* Giovanni Rombolà,* Riccardo Saccardi,*Letizia Lombardini,* Nicola Di Lorenzo,§ Gabriella B. Vannelli,¶ and Pasquale Gallina

*Careggi University Hospital, Florence, Italy
†Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy
‡Department of Neurosciences, University of Florence, Florence, Italy
§Department of Surgery and Translational Medicine, University of Florence, Florence, Italy
¶Department of Clinical and Experimental Medicine, University of Florence, Florence, Italy

Fetal grafting in a human diseased brain was thought to be less immunogenic than other solid organ transplants, hence the minor impact on the efficacy of the transplant. How much prophylactic immune protection is required for neural allotransplantation is also debated. High-sensitive anti-HLA antibody screening in this field has never been reported. Sixteen patients with Huntington’s disease underwent human fetal striatal transplantation in the frame of an open-label observational trial, which is being carried out at Florence University. All patients had both brain hemispheres grafted in two separate robotic–stereotactic procedures. The trial started in February 2006 with the first graft to the first patient (R1). R16 was given his second graft on March 2011. All patients received triple immunosuppressive treatment. Pre- and posttransplant sera were analyzed for the presence of anti-HLA antibodies using the multiplexed microsphere-based suspension array Luminex xMAP technology. Median follow-up was 38.5 months (range 13–85). Six patients developed anti-HLA antibodies, which turned out to be donor specific. Alloimmunization occurred in a time window of 0–49 months after the first neurosurgical procedure. The immunogenic determinants were non-self-epitopes from mismatched HLA antigens. These determinants were both public epitopes shared by two or more HLA molecules and private epitopes unique to individual HLA molecules. One patient had non-donor-specific anti-HLA antibodies in herpretransplant serum sample, possibly due to previous sensitization events. Although the clinical significance of donor-specific antibodies is far from being established, particularly in the setting of neuronal transplantation, these findings underline the need of careful pre- and posttransplant immunogenetic evaluation of patients with intracerebral grafts.

Key words: Donor-specific antibodies; Fetal striatal tissue; Huntington’s disease; Neural transplantation

Received September 19, 2013; final acceptance December 13, 2013. Online prepub date: December 30, 2013.
Address correspondence to Berardino PorfirioImmunogenetics Laboratory, Clinical Physiopathology Unit, Department of Clinical and Experimental Biomedical Sciences, University of Florence, Viale G. Pieraccini, 6 50139 Florence, Italy. Tel: +39-055-427-1481; Fax: +39-055-427-1413; E-mail: nporfirio@unifi.it


Cell Transplantation, Vol. 24, pp. 819-828, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679336
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Differential Clearance of Rat and Human Bone Marrow-Derived Mesenchymal Stem Cells From the Brain After Intra-arterial Infusion in Rats

Joonas Khabbal,*1 Erja Kerkelä,†1 Bhimashankar Mitkari,*1 Mari Raki,‡ Johanna Nystedt,† Ville Mikkonen,* Kim Bergström,‡§ Saara Laitinen,† Matti Korhonen,† and Jukka Jolkkonen*

*Institute of Clinical Medicine – Neurology, University of Eastern Finland, Kuopio, Finland
†Finnish Red Cross Blood Service, Advanced Therapies and Product Development, Helsinki, Finland
‡Centre for Drug Research, Faculty of Pharmacy, University of Helsinki, Helsinki, Finland
§HUS Medical Imaging Centre, Helsinki University Central Hospital, Helsinki, Finland

Intra-arterial (IA) delivery of bone marrow-derived mesenchymal stem cells (BM-MSCs) has shown potential as a minimally invasive therapeutic approach for stroke. The aim of the present study was to determine the whole-body biodistribution and clearance of technetium-99m (99mTc)-labeled rat and human BM-MSCs after IA delivery in a rat model of transient middle cerebral artery occlusion (MCAO) using single-photon emission computed tomography (SPECT). Our hypothesis was that xenotransplantation has a major impact on the behavior of cells. Male RccHan:Wistar rats were subjected to sham operation or MCAO. Twenty-four hours after surgery, BM-MSCs (2 × 106
cells/animal) labeled with 99mTc were infused into the external carotid artery. Whole-body SPECT images were acquired 20 min, 3 h, and 6 h postinjection, after which rats were sacrificed, and organs were collected and weighed for measurement of radioactivity. The results showed that the majority of the cells were located in the brain and especially in the ipsilateral hemisphere immediately after cell infusion both in sham-operated and MCAO rats. This was followed by fast disappearance, particularly in the case of human cells. At the same time, the radioactivity signal increased in the spleen, kidney, and liver, the organs responsible for destroying cells. Further studies are needed to demonstrate whether differential cell behavior has any functional impact.

Key words: Intra-arterial (IA) delivery; Biodistribution; Bone marrow-derived mesenchymal cells (BM-MSCs); Cerebral ischemia; Single-photon emission computed tomography (SPECT) imaging; Xenografting

Received August 30, 2013; final acceptance February 11, 2014. Online prepub date: March 3, 2014.
1These authors provided equal contribution to this work. Address correspondence to Jukka Jolkkonen, Ph.D., Institute of Clinical Medicine – Neurology, University of Eastern Finland, Yliopistonranta 1 C, 70210 Kuopio, Finland. Tel: +358-40-3552519; Fax: +358-17-162048; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 829-844, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675179
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Prerequisite OCT4 Maintenance Potentiates the Neural Induction of Differentiating Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

Sheng-Mei Chen,* Maw-Sheng Lee,†‡§¶# Chia-Yu Chang,** Shinn-Zong Lin,††‡‡§§ En-Hui Cheng,# Yung-Hsien Liu,¶¶ Hong-Chuan Pan,## Hsiu-Chin Lee,** and Hong-Lin Su***

*Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, Taichung, Taiwan
†Institute of Medicine, China Medical University, Taichung, Taiwan
‡Department of Obstetrics and Gynecology, China Medical University, Taichung, Taiwan
§Department of Obstetrics and Gynecology, Chung-Shan Medical University Hospital, Taichung, Taiwan
¶Department of Medicine, Chung-Shan Medical University, Taichung, Taiwan
#Division of Infertility, Lee Women’s Hospital, Taichung, Taiwan
**Ph.D. Program in Tissue Engineering and Regenerative Medicine, Agricultural Biotechnology Center, National Chung Hsing University, Taichung, Taiwan
††Graduate Institute of Immunology, China Medical University, Taichung, Taiwan
‡‡Center for Neuropsychiatry, China Medical University and Hospital, Taichung, Taiwan
§§Department of Neurosurgery, China Medical University Beigan Hospital, Yunlin, Taiwan
¶¶Profertile IVF Center, Kaohsiung, Taiwan
##Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan

Establishing an efficient differentiation procedure is prerequisite for the cell transplantation of pluripotent stem cells. Activating fibroblast growth factor (FGF) signals and inhibiting the activin/nodal pathway are both conserved principles to direct the neural induction (NI) of developing embryos and human embryonic stem cells (hESCs). Wnt signal and OCT4 expression are critical for the hESC pluripotency; however, their roles in cell differentiation are largely unclear. We demonstrate that in the presence of FGF2 and activin inhibitor SB431542, applying a small-molecule Wnt agonist, BIO, efficiently and rapidly steers the NI of all our tested hESCs. A human induced pluripotent stem cell (iPSC), which is refractory for efficient neural conversion by FGF2, effectively differentiated to SOX1+
cells after the BIO/SB431542/FGF2 treatment. In addition, BIO promoted cell survival and transiently sustained OCT4 expression at the early NI stage with FGF2 and SB431542. Interestingly, at the late NI stage, the OCT4 level rapidly declined in the treated hESCs and consequently initiated the formation of neural rosettes with forebrain neuron characteristics. This study illustrates the distinct effects of Wnt activation on maintaining pluripotency and committing neural lineages at the early and late NI stages of hESCs and iPSCs, respectively.

Key words: Embryonic stem cells; Neural induction (NI); Neural progenitor cells; Wnt

Received October 2, 2013; final acceptance November 8, 2013. Online prepub date: November 20, 2013.
Address correspondence to Hong-Lin Su, Department of Life Sciences, National Chung Hsing University, 250 Kuo-Kuang Road, Taichung 402, Taiwan. Tel: +886-422840416; Fax: +886-422854391; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 845-864, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X674639
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Characteristic Expression of Major Histocompatibility Complex and Immune Privilege Genes in Human Pluripotent Stem Cells and Their Derivatives

Hsin-Fu Chen,*†‡ Chun-Ying Yu,§ Mei-Jou Chen,* Shiu-Huey Chou,¶ Ming-Shan Chiang,* Wen-Hsi Chou,* Bor-Sheng Ko,#** Hsiang-Po Huang,‡ Hung-Chih Kuo,§1 and Hong-Nerng Ho*†††1

*Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, College of Medicine and the Hospital, National Taiwan University, Taipei, Taiwan
†Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan
‡Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan
§Institute of Cellular and Organismic Biology and Genomics Research Center, Academia Sinica, Taipei, Taiwan
¶Department of Life Science, Fu-Jen Catholic University, Taipei, Taiwan
#Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan
**Institute of Cellular and Systemic Medicine, National Health Research Institute, Miaoli County, Taiwan
††Graduate Institute of Immunology, College of Medicine, National Taiwan University, Taipei, Taiwan

Pluripotent stem cells, including human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs), have been regarded as useful sources for cell-based transplantation therapy. However, immunogenicity of the cells remains the major determinant for successful clinical application. We report the examination of several hESC lines (NTU1 and H9), hiPSC lines, and their derivatives (including stem cell-derived hepatocytes) for the expression of major histocompatibility complex (MHC), natural killer (NK) cell receptor (NKp30, NKp44, NKp46) ligand, immune-related genes, human leukocyte antigen (HLA) haplotyping, and the effects in functional mixed lymphocyte reaction (MLR). Flow cytometry showed lower levels (percentages and fluorescence intensities) of MHC class I (MHC-I) molecules, β2-microglobulin, and HLA-E in undifferentiated stem cells. The levels were increased after cotreatment with interferon-γ and/or in vitro differentiation. Antigen-presenting cell markers (CD11c, CD80, and CD86) and MHC-II (HLA-DP, -DQ, and -DR) remained low throughout the treatments. Recognition of stem cells/derivatives by NK lysis receptors were lower or absent. Activation of responder lymphocytes was significantly lower by undifferentiated stem cells than by allogeneic lymphocytes in MLR, but differentiated NTU1 hESCs induced a cell number-dependent lymphocyte proliferation comparable with that by allogeneic lymphocytes. Interestingly, activation of lymphocytes by differentiated hiPSCs or H9 cells became blunted at higher cell numbers. Real-time reverse transcriptase PCR (RT-PCR) showed significant differential expression of immune privilege genes (TGF-β2Arginase 2Indole 1GATA3POMCVIPCALCA,CALCBIL-1RNCD95LCR1LSerpine 1HMOX1IL6LGALS3HEBP1THBS1CD59, and LGALS1) in pluripotent stem cells/derivatives when compared to somatic cells. It was concluded that pluripotent stem cells/derivatives are predicted to be immunogenic, though evidence suggests some level of potential immune privilege. In addition, differential immunogenicity may exist between different pluripotent stem cell lines and their derivatives.

Key words: Pluripotent stem cells; Embryonic stem cells; Induced pluripotent stem cells; Immunogenicity; Major histocompatibility complex (MHC); Immune privilege

Received April 18, 2012; final acceptance October 9, 2013. Online prepub date: October 18, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Hong-Nerng Ho, M.D., National Taiwan University Hospital, Department of Obstetrics and Gynecology, No.7 Chung-Shan S. Rd., Taipei 100, Taiwan. Tel: +886-2-23123456, #71501; Fax: +886-2-23116056; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Hung-Chih Kuo, Ph.D., Stem Cell Program, Genomics Research Center/Institute of Cellular and Organismic Biology, Academia Sinica, No. 128, Sec. 2, Academia Road, Nankang, Taipei 11529, Taiwan. Tel: +886 2 27899580, ext. 201; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 865-877, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676231
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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The Human Umbilical Cord Tissue-Derived MSC Population UCX® Promotes Early Motogenic Effects on Keratinocytes and Fibroblasts and G-CSF-Mediated Mobilization of BM-MSCs When Transplanted In Vivo

Joana P. Miranda,*1 Elysse Filipe,*1 Ana Sofia Fernandes,*† Joana M. Almeida,‡ José P. Martins,‡ Alexandre De la Fuente,§ Miguel Abal,§ Rita N. Barcia,‡ Pedro Cruz,‡ Helder Cruz,‡ Matilde Castro,* and Jorge M. Santos‡

*Research Institute for Medicines, Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal
†Centro de Biociências da Universidade Lusófona (CBios), Lisbon, Portugal
ECBio S.A., Amadora, Portugal
§Translational Medical Oncology, Health Research Institute of Santiago (IDIS), Complexo Hospitalario Universitario de Santiago de Compostela/SERGAS, Santiago de Compostela, Spain

Mesenchymal stromal cells (MSCs) play an important role in tissue regeneration mainly through the secretion of trophic factors that enhance the repair of damaged tissues. The main goal of this work was to study the paracrine mechanisms by which an umbilical cord tissue-derived MSC population (UCX®) promotes the migration capacity of human dermal fibroblasts and keratinocytes, which is highly relevant for skin regeneration. Furthermore, the differences between paracrine activities of MSCs from the umbilical cord tissue and the bone marrow (BM-MSCs) were also evaluated. In vitro scratch assays revealed that conditioned media (CM) obtained from both growing and stationary-phase UCX
® cultures induced human dermal fibroblast (HDF) and keratinocyte (HaCaT) migration. These assays showed that the motogenic activity of UCX® CM to HaCaTs was significantly higher than to HDFs, in opposition to the effect seen with CM produced by BM-MSCs that preferentially induced HDF migration. Accordingly, a comparative quantification of key factors with vital importance in the consecutive stages of wound healing revealed very different secretome profiles between UCX® and BM-MSCs. The relatively higher UCX® expression of EGF, FGF-2, and KGF strongly supports early induction of keratinocyte migration and function, whereas the UCX®-specific expression of G-CSF suggested additional roles in mobilization of healing-related cells including CD34/CD45 precursors (MSCs) known to be involved in tissue regeneration. Accordingly, in vitro chemotaxis assays and an in vivo transplantation model for chemoattraction confirmed that UCX® are chemotactic to CD34/CD45 BM-MSCs via a cell-specific mobilization mechanism mediated by G-CSF. Overall, the results strongly suggest different paracrine activities between MSCs derived from different tissue sources, revealing the potential of UCX® to extend the regenerative capacity of the organism by complementing the role of endogenous BM-MSCs.

Key words: Mesenchymal stromal cells (MSCs); UCX®; Stem cell transplantation; Fibroblast and keratinocyte migration; BM-MSC mobilization; G-CSF; Skin regeneration

Received July 19, 2013; final acceptance December 17, 2013. Online prepub date: December 30, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Jorge Miguel Santos, ECBio S.A., Rua Henrique Paiva Couceiro, Nº 27, 2700-451 Amadora, Portugal. Tel: +351 214997579; Fax: +351 214997594; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 879-890, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676899
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Two Distinct Mechanisms Mediate the Involvement of Bone Marrow Cells in Islet Remodeling: Neogenesis of Insulin-Producing Cells and Support of Islet Recovery

Svetlana Iskovich,* Nitza Goldenberg-Cohen,† Tamila Sadikov,*† Isaac Yaniv,‡ Jerry Stein,§ and Nadir Askenasy*

*Frankel Laboratory, Schneider Children’s Medical Center of Israel, Petach Tikva, Israel
†Krieger Eye Research Laboratory, Center for Stem Cell Research, Schneider Children’s Medical Center of Israel, Petach Tikva, Israel
‡Department of Pediatric Hematology-Oncology, Schneider Children’s Medical Center of Israel, Petach Tikva, Israel
§Bone Marrow Transplant Unit, Schneider Children’s Medical Center of Israel, Petach Tikva, Israel

We have recently reported that small-sized bone marrow cells (BMCs) isolated by counterflow centrifugal elutriation and depleted of lineage markers (Fr25lin) have the capacity to differentiate and contribute to regeneration of injured islets. In this study, we assess some of the characteristics of these cells compared to elutriated hematopoietic progenitors (R/O) and whole BMCs in a murine model of streptozotocin-induced chemical diabetes. The GFPbrightCD45+
progeny of whole BMCs and R/O progenitors progressively infiltrate the pancreas with evolution of donor chimerism; are found at islet perimeter, vascular, and ductal walls; and have a modest impact on islet recovery from injury. In contrast, Fr25lin cells incorporate in the islets, convert to GFPdimCD45PDX-1+ phenotypes, produce proinsulin, and secrete insulin with significant contribution to stabilization of glucose homeostasis. The elutriated Fr25lin cells express low levels of CD45 and are negative for SCA-1 and c-kit, as removal of cells expressing these markers did not impair conversion to produce insulin. BMCs mediate two synergistic mechanisms that contribute to islet recovery from injury: support of islet remodeling by hematopoietic cells and neogenesis of insulin-producing cells from stem cells.

Key words: Chemical diabetes; Islet remodeling; β-Cell regeneration; Insulin-producing cells; Bone marrow stem cells; Hematopoietic progenitors; Bone marrow transplantation

Received June 12, 2013; final acceptance December 18, 2013. Online prepub date: December 30, 2013.
Address correspondence to Nadir Askenasy, M.D., Ph.D., Frankel Laboratory, Center for Stem Cell Research, Schneider Children’s Medical Center of Israel, 14 Kaplan Street, Petach Tikva, Israel 49202. Tel: +972 3921 3954; Fax: +972 3921 4156; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 891-907, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X681919
E-ISSN 1555-3892
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Engineered Hair Follicle Mesenchymal Stem Cells Overexpressing Controlled-Release Insulin Reverse Hyperglycemia in Mice With Type l Diabetes

Chunling Wu,* Feilin Liu,* Pengdong Li,* Guifang Zhao,* Shaowei Lan,* Wenyue Jiang,† Xiangwei Meng,‡ Lixing Tian,§ Gang Li,¶ Yulin Li,* and Jin Yu Liu*

*The Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, P.R. China
†School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin, P.R. China
‡Department of Gastroenterology, Fist Hospital of Jilin University, Changchun, Jilin, P.R. China
§Department of Pathology, Changchun Obstetrics and Gynecology Hospital, Changchun, Jilin, P.R. China
¶Department of Orthopaedics and Traumatology, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, P.R. China

Genetically engineered stem cells that overexpress genes encoding therapeutic products can be exploited to correct metabolic disorders by repairing and regenerating diseased organs or restoring their function. Hair follicles are readily accessible and serve as a rich source of autologous stem cells for cell-based gene therapy. Here we isolated mesenchymal stem cells from human hair follicles (HF-MSCs) and engineered them to overexpress the human insulin gene and release human insulin in a time- and dose-dependent manner in response to rapamycin. The engineered HF-MSCs retained their characteristic cell surface markers and retained their potential to differentiate into adipocytes and osteoblasts. When mice with streptozotocin-induced type 1 diabetes were engrafted with these engineered HF-MSCs, these cells expressed and released a dose of human insulin, dramatically reversed hyperglycemia, and significantly reduced death rate. Moreover, the engineered HF-MSCs did not form detectable tumors throughout the 120-day animal tests in our experiment. Our results show that HF-MSCs can be used to safely and efficiently express therapeutic transgenes and therefore show promise for cell-based gene therapy of human disease.

Key words: Mesenchymal stem cells; Hair follicles; Gene therapy; Lentivirus; Type 1 diabetes

Received December 18, 2013; final acceptance May 10, 2014. Online prepub date: May 15, 2014.
Address correspondence to Jin Yu Liu, The Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin 130021, P.R. China. Tel: +086-431-85619481; Fax: +086-431-85619143; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or YulinLi, The Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin 130021, P.R. China. E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 909-920, 2015
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DOI: http://dx.doi.org/10.3727/096368913X676907
E-ISSN 1555-3892
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Bone Morphogenetic Protein 9 and 13 Induce C3H10T1/2 Cell Differentiation to Cardiomyocyte-Like Cells In Vitro

Lu Chen,*† Hao Wei,‡ Jintong Tan,§ Hengsheng Chen,† Zhenguo Liu,* and Yuan Chen*

*Department of Cardiology, Children’s Hospital of Chongqing Medical University, Chongqing, China
†Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Children’s Hospital of Chongqing Medical University, Chongqing, China
‡Department of Pediatrics, Hospital of North Sichuan Medical College, Nanchong, China
§Department of Pediatrics, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

The present study aimed to evaluate the effect of bone morphogenetic protein 9 (BMP9) and BMP13 on cardiac differentiation of C3H10T1/2 cells in vitro and to characterize the differentiated cells on their ultrastructure and transmembrane electrophysiological features. C3H10T1/2 cells were transfected with the vectors for BMP9 or BMP13 and differentiated into cardiomyocytes in vitro for up to 28 days. The expression of cardiac-specific genes Gata4 and Mef2c and proteins troponin T (cTnT) and connexin 43 (Cx43) was significantly increased in the cells transfected with BMP9 or BMP13 after differentiation over the controls as evaluated using quantitative RT-PCR, Western blotting, and immunofluorescence staining. Transmission electron microscopy and Masson trichrome staining showed that the specific myocardial leap dish and myofilament-like structure were present in the cells overexpressing BMP9 or BMP13, not in the control cells. Whole-cell patch-clamping study demonstrated the presence of delayed rectifier potassium current, inward rectifier potassium current, and T-type calcium current in the cells overexpressing BMP9 or BMP13. Sodium current was detected in a small number of cells overexpressing BMP9, not in the BMP13-transfected cells or the control cells. The expression of Mef2c gene and Cx43 and cTnT proteins was also significantly higher in the cells overexpressing BMP9 than those overexpressing BMP13. Our data indicate that BMP9 and BMP13 (BMP9 might be more effective) promoted the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells with cellular ultrastructures and ion channel currents similar to mature cardiomyocytes in vitro.

Key words: Bone morphogenetic protein 9 (BMP9); Bone morphogenetic protein 13 (BMP13); C3H10T1/2 cells; Cardiomyocytes; Ion channel current

Received July 8, 2013; final acceptance December 19, 2013. Online prepub date: December 30, 2013.
Address correspondence to Yuan Chen, M.D., Department of Cardiology, the Children’s Hospital of Chongqing Medical University, 136 Zhongshan Er Road, Yu Zhong District, Chongqing 400014, China. Tel: +86 023 63631938; Fax: +86 023 63622874; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Zhenguo Liu, M.D., Ph.D., Department of Cardiology, the Children’s Hospital of Chongqing Medical University, 136 Zhongshan Er Road, Yu Zhong District, Chongqing 400014, China. E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 921-937, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X678580
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Transplantation of Human Dental Pulp-Derived Stem Cells Protects Against Heatstroke in Mice

Ling-Shu Tseng,* Sheng-Hsien Chen,†‡§ Mao-Tsun Lin,†§ and Ying-Chu Lin*

*School of Dentistry, Kaohsiung Medical University, Kaohsiung City, Taiwan
†Department of Medical Research, Chi Mei Medical Center, Tainan City, Taiwan
‡Da-An Women and Children Hospital, Tainan, Taiwan
§Department of Biotechnology, Southern Taiwan University of Science and Technology, Tainan, Taiwan

Stem cells from human exfoliated deciduous tooth pulp (SHED) is a promising approach for the treatment of stroke and spinal cord injury. In this study, we investigated the therapeutic effects of SHED for the treatment of multiple organ (including brain, particularly hypothalamus) injury in heatstroke mice. ICR male mice were exposed to whole body heating (WBH; 41.2°C, relative humidity 50–55%, for 1 h) and then returned to normal room temperature (26°C). We observed that intravenous administration of SHED immediately post-WBH exhibited the following therapeutic benefits for recovery after heatstroke: (a) inhibition of WBH-induced neurologic and thermoregulatory deficits; (b) reduction of WBH-induced ischemia, hypoxia, and oxidative damage to the brain (particularly the hypothalamus); (c) attenuation of WBH-induced increased plasma levels of systemic inflammatory response molecules, such as tumor necrosis factor-α and intercellular adhesion molecule-1; (d) improvement of WBH-inducedhypothalamo–pituitary–adrenocortical (HPA) axis activity (as reflected by enhanced plasma levels of both adrenocorticotrophic hormone and corticosterone); and (e) attenuation of WBH-induced multiple organ apoptosis as well as lethality. In conclusion, post-WBH treatment with SHED reduced induction of proinflammatory cytokines and oxidative radicals, enhanced plasma induction of both adrenocorticotrophic hormone and corticosterone, and improved lethality in mouse heatstroke. The protective effect of SHED may be related to a decreased inflammatory response, decreased oxidative stress, and an increased HPA axis activity following the WBH injury.

Key words: Heatstroke; Human exfoliated deciduous teeth pulp stem cells; Brain; Thermoregulation; Neurologic severity scores; Multiple organs

Received July 16, 2013; final acceptance January 29, 2014. Online prepub date: March 7, 2014.
Address correspondence to Dr. Ying-Chu Lin, Ph.D., School of Dentistry, Kaohsiung Medical University, Kaohsiung City 807, Taiwan. Tel: 866 7 3121101, ext. 2753; Fax: 866 7 3157024; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 939-953, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X678599
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Therapeutic Effects of Mouse Adipose-Derived Stem Cells and Losartan in the Skeletal Muscle of Injured Mdx Mice

Eun-Mi Lee,*† Ah-Young Kim,*† Eun-Joo Lee,*† Jin-Kyu Park,* Myeong-Mi Lee,*† Meeyul Hwang,*† Choong-Yong Kim,‡ Shin-Yoon Kim,§¶ and Kyu-Shik Jeong*†

*College of Veterinary Medicine, Kyungpook National University, Daegu City, Republic of Korea
†Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu City, Republic of Korea
‡Animal Center, Daegu-Gueongbuk Medical Innovation Foundation, Daegu City, Republic of Korea
§Department of Orthopedic Surgery, Kyungpook National University School of Medicine, Deagu City, Republic of Korea
¶Skeletal Diseases Genome Research Center, Kyungpook National University and Hospital, Deagu City, Republic of Korea

Duchenne muscular dystrophy (DMD) is an X-linked genetic disorder caused by mutations in the dystrophin gene. Adipose-derived stem cells (ASCs) are an attractive source of cells for stem cell therapy. Losartan has been reported to improve ASC transplantation in injured mouse muscles. In the present study, we investigated whether the combined treatment of losartan and ASCs in the injured muscles of mdx mice improves regeneration. The combined treatment of ASCs and losartan remarkably improved muscle regeneration and induced muscle hypertrophy. In addition, ASCs and losartan treatment downregulated transforming growth factor-β and inhibited muscle fibrosis. We observed cells coexpressing green fluorescent protein (GFP) and dystrophin in the muscle samples of mice transplanted with GFP-positive ASCs. In the coculture in vitro experiment, we also observed that the GFP ASCs differentiated into dystrophin-expressing myotubes. The present study shows that the combination of transplanted ASCs and treatment with losartan ameliorated muscle fibrosis and improved muscle regeneration in injured mdx mice. Thus, we suggest that combined treatment with losartan and ASCs could help to improve muscle regeneration in the muscles of injured patients, including DMD patients.

Key words: Adipose stem cells; Losartan; Stem cell microenvironment; Muscular dystrophy; Skeletal muscle regeneration

Received January 20, 2012; final acceptance January 27, 2014. Online prepub date: March 3, 2014.
Address correspondence to Kyu-Shik Jeong, D.V.M., Ph.D., Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea, 702-701. Tel: +82-53-950-5975; Fax: +82-53-950-5955; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it