Cell Transplantation 21(8) Abstracts

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Cell Transplantation, Vol. 21, pp. 1603–1611, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X647234
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Teratocarcinoma Formation in Embryonic Stem Cell-Derived Neural Progenitor Hippocampal Transplants

Noélle D. Germain,* Nathaniel W. Hartman,*1 Chunyu Cai,*2 Sandy Becker,* Janice R. Naegele,*† and Laura B. Grabel*

*Department of Biology, Wesleyan University, Middletown, CT, USA
†Program in Neuroscience and Behavior, Wesleyan University, Middletown, CT, USA

Embryonic stem cells (ESCs) hold great therapeutic potential due to their ability to differentiate into cells of the three primary germ layers, which can be used to repopulate disease-damaged tissues. In fact, two cell therapies using ESC derivatives are currently in phase I clinical trials. A main concern in using ESCs and their derivatives for cell transplantation is the ability of undifferentiated ESCs to generate tumors in the host. Positive selection steps are often included in protocols designed to generate particular cell types from ESCs; however, the transition from ESC to progenitor cell or terminally differentiated cell is not synchronous, and residual undifferentiated cells often remain. In our transplants of ESC-derived neural progenitors (ESNPs) into the adult mouse hippocampus, we have observed the formation of teratocarcinomas. We set out to reduce teratocarcinoma formation by enrichment of ESNPs using fluorescence-activated cell sorting (FACS) and have found that, although enrichment prior to transplant reduces the overall rate of teratocarcinoma formation, the tumorigenicity of cell batches can vary widely, even after FACS enrichment to as much as 95% ESNPs. Our data suggest that this variability may be due to the percentage of residual ESCs remaining in the transplant cell population and to the presence of pluripotent epiblast-like cells, not previously identified in transplant batches. Our data emphasize the need for stringent characterization of transplant cell populations that will be used for cell replacement therapies in order to reduce the risk of tumor formation.

Key words: Embryonic stem cells; Hippocampus; Teratocarcinoma; Neural progenitor

Received April 22, 2011; final acceptance September 22, 2011. Online prepub date: July 5, 2012.
1Current address: Department of Neurosurgery, Yale University School of Medicine, New Haven, CT, USA.
2Current address: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.
Address correspondence to Laura B. Grabel, Department of Biology, Hall-Atwater Laboratories, Wesleyan University, Middletown, CT 06459, USA. Tel: +1 860 685 3238; Fax: +1 860 685 3279; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1613–1627, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640475
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bystander Effect on Brain Tissue of Mesoangioblasts Producing Neurotrophins

Tao Su,*†1 Raffaella Scardigli,‡§1 Luisa Fasulo,§1 Beatrice Paradiso,*† Mario Barbieri,*† Anna Binaschi,*† Roberta Bovolenta,*† Silvia Zucchini,*†¶ Giulio Cossu,# Antonino Cattaneo,§**2 and Michele Simonato*†¶2

*Department of Clinical and Experimental Medicine, Section of Pharmacology, Neuroscience Center, University of Ferrara, Ferrara, Italy
†National Institute of Neuroscience, Ferrara Italy
‡Istituto di Farmacologia Traslazionale, CNR, Rome, Italy
§European Brain Research Institute, Rome, Italy
¶Tecnopolo di Ferrara, Laboratorio per le Tecnologie delle Terapie Avanzate, Ferrara, Italy
#Division of Regenerative Medicine, San Raffaele Scientific Institute, Milan, Italy
**Scuola Normale Superiore, Pisa, Italy

Neurotrophic factors (NTFs) are involved in the regulation of neuronal survival and function and, thus, may be used to treat neurological diseases associated with neuronal death. A major hurdle for their clinical application is the delivery mode. We describe here a new strategy based on the use of progenitor cells called mesoangioblasts (MABs). MABs can be isolated from postnatal mesoderm tissues and, because of a high adhesin-dependent migratory capacity, can reach perivascular targets especially in damaged areas. We generated genetically modified MABs producing nerve growth factor (MABs-NGF) or brain-derived neurotrophic factor (MABs-BDNF) and assessed their bystander effects in vitro using PC12 cells, primary cultures, and organotypic cultures of adult hippocampal slices. MABs-NGF-conditioned medium induced differentiation of PC12 cells, while MABs-BDNF-conditioned medium increased viability of cultured neurons and slices. Slices cultured with MABs-BDNF medium also better retained their morphology and functional connections, and all these effects were abolished by the TrkB kinase blocker K252a or the BDNF scavenger TrkB-IgG. Interestingly, the amount of BDNF released by MABs-BDNF produced greater effects than an identical amount of recombinant BDNF, suggesting that other NTFs produced by MABs synergize with BDNF. Thus, MABs can be an effective vehicle for NTF delivery, promoting differentiation, survival, and functionality of neurons. In summary, MABs hold distinct advantages over other currently evaluated approaches for NTF delivery in the CNS, including synergy of MAB-produced NTF with the neurotrophins. Since MABs may be capable of homing into damaged brain areas, they represent a conceptually novel, promising therapeutic approach to treat neurodegenerative diseases.

Key words: Mesoangioblast; Neurotrophin; Neurodegeneration; Stem cells

Received March 4, 2011; final acceptance October 17, 2011. Online prepub date: April 17, 2012.
1These authors provided equal contribution to this work.
2These authors share the position of senior author.
Address correspondence to Michele Simonato, M.D., Neuroscience Center, University of Ferrara, via Fossato di Mortara 17–19, 44100 Ferrara, Italy. Tel: +39-0532-455211; Fax: +39-0532-455205; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1629–1640, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640556
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Mesenchymal Stem Cells Augment Neurogenesis in the Subventricular Zone and Enhance Differentiation of Neural Precursor Cells Into Dopaminergic Neurons in the Substantia Nigra of a Parkinsonian Model

Hyun-Jung Park,*† Jin Young Shin,*† Bo Ra Lee,*† Hyun Ok Kim,‡ and Phil Hyu Lee*†

*Severance Biomedical Science Institute, Yonsei University, Seoul, South Korea
†Department of Neurology, Yonsei University College of Medicine, Seoul, South Korea
‡Department of Laboratory Medicine, Yonsei Cell Therapy Center, Yonsei University College of Medicine, Seoul, South Korea

Growing evidence has demonstrated that neurogenesis in the subventricular zone (SVZ) is significantly decreased in Parkinson’s disease (PD). Modulation of endogenous neurogenesis would have a significant impact on future therapeutic strategies for neurodegenerative diseases. In the present study, we investigated the augmentative effects of human mesenchymal stem cells (hMSCs) on neurogenesis in a PD model. Neurogenesis was assessed in vitro with 1-methyl-4-phenylpyridinium (MPP+) treatment using neural precursor cells (NPCs) isolated from the SVZ and in vivo with a BrdU-injected animal model of PD using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Immunochemical analyses were used to measure neurogenic activity. The number of BrdU-ir cells in the SVZ and the substantia nigra (SN) was significantly increased in the hMSC-treated PD group compared with the MPTP-only-treated group. Double-stained cells for BrdU and tyrosine hydroxylase were notably observed in the SN of hMSC-treated PD animals, and they did not colocalize with the nuclear matrix; however, double-stained cells were not detected in the SN of the MPTP-induced PD animal model. Furthermore, hMSC administration increased the expression of the epidermal growth factor receptor (EGFR) in the SVZ of PD animals, and the coculture of hMSCs significantly increased the release of EGF in the medium of MPP+-treated NPCs. The present study demonstrated that hMSC administration significantly augmented neurogenesis in both the SVZ and SN of PD animal models, which led to increased differentiation of NPCs into dopaminergic neurons in the SN. Additionally, hMSC-induced modulation of EGF seems to be an underlying contributor to the enhancement of neurogenesis by hMSCs. The modulation of endogenous adult neurogenesis to repair the damaged PD brain using hMSCs would have a significant impact on future strategies for PD treatment.

Key words: Mesenchymal stem cells; Neurogenesis; Parkinson’s disease

Received March 21, 2011; final acceptance October 15, 2011. Online prepub date: April 24, 2011.
Address correspondence to Phil Hyu Lee, M.D., Ph.D., Department of Neurology, Yonsei University College of Medicine, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-752, South Korea. Tel: +82-2-2228-1608; Fax: +82-2-393-0705; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1641–1650, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637488
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Mesenchymal Stem Cells Overexpressing Hepatocyte Growth Factor (HGF) Inhibit Collagen Deposit and Improve Bladder Function in Rat Model of Bladder Outlet Obstruction

Yun Seob Song,*1 Hong Jun Lee,†‡1 Seung Hwan Doo,* Sun Ju Lee,§ Inja Lim,¶ Kyu-Tae Chang,# and Seung U. Kim†‡

*Department of Urology, Soonchunhyang University School of Medicine, Seoul, South Korea
†Medical Research Institute, Chung-Ang University College of Medicine, Seoul, South Korea
‡Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, Canada
§Department of Medicine, Kyunghee University School of Medicine, Seoul, South Korea
¶Department of Physiology, Chung-Ang University College of Medicine, Seoul, South Korea
#National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, South Korea

Bladder outlet obstruction (BOO) caused by collagen deposit is one of the most common problems in elderly male. This study was performed to examine the capability of human mesenchymal stem cells (MSCs) overexpressing hepatocyte growth factor (HGF) to inhibit collagen deposition in rat model of bladder outlet obstruction (BOO). HGF is known for its antifibrotic effect and the most promising agent for treating bladder fibrosis. BM3.B10 stable immortalized human MSC line (B10) was transduced to encode human HGF with a retroviral vector was prepared (B10.HGF). Two weeks after the onset of BOO, B10, and B10.HGF cells were injected into the rat’s bladder wall. After 4 weeks, bladder tissues were harvested and Masson’s trichrome staining was performed. Transgene expression in HGF-expressing B10 cells was demonstrated by reverse transcriptase polymerase chain reaction and immunohistochemical staining, and the high levels of HGF secreted by B10. HGF cells was confirmed by ELISA. The mean bladder weight in BOO rats was 5.8 times of the normal controls, while in animals grafted with B10.HGF cells, the weight was down to four times of the control [90.2 ± 1.6 (control), 89.9 ± 2.8 (sham), 527.9 ± 150.9 (BOO), 447.7 ± 41.0 (BOO + B10), and 362.7 ± 113.2 (BOO + B10.HGF)]. The mean percentage of collagen area increased in BOO rats, while in the animals transplanted with B10.HGF cells, the collagen area decreased to the normal control level [12.2 ± 1.3, (control), 12.8 ± 1.1 (sham), 26.6 ± 2.7 (BOO), 19.9 ± 6.0 (BOO + B10), and 13.3 ± 2.1 (BOO + B10.HGF)]. The expression of collagen and TGF-β protein increased after BOO, while the expression of HGF and c-met protein increased in the group with B10.HGF transplantation after BOO. Intercontraction interval decreased after BOO, but it recovered after B10.HGF transplantation. Maximal voiding pressure (MVP) increased after BOO, and it recovered to levels of the normal control after transplantation of B10.HGF cells. Residual urine volume (RU) increased after BOO, but the RU increase was not reversed by transplantation of B10.HGF cells. Human MSCs overexpressing HGF inhibited collagen deposition and improved cystometric parameters in bladder outlet obstruction of rats. The present study indicates that transplantation of MSCs modified to overexpress HGF could serve as a novel therapeutic strategy against bladder fibrosis in patients with bladder outlet obstruction.

Key words: Bladder outlet obstruction; Bladder fibrosis; Collagen deposit; Human mesenchymal stem cells; Cell transplantation; Hepatocyte growth factor

Received May 25, 2011; final acceptance October 15, 2011. Online prepub date: April 10, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Seung U. Kim, M.D., Ph.D., Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, BC V6T2B5, Canada. Tel: +1-604-822-7145; Fax: +1-604-822-7897; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1651–1665, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X647234
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Therapeutic Potential of Anterior Cruciate Ligament-Derived Stem Cells for Anterior Cruciate Ligament Reconstruction

Yutaka Mifune,*†‡ Tomoyuki Matsumoto,*†‡ Shusuke Ota,*† Makoto Nishimori,*† Arvydas Usas,*† Sebastian Kopf,*† Ryosuke Kuroda,‡ Masahiro Kurosaka,‡ Freddie H. Fu,† and Johnny Huard*†§

*Stem Cell Research Center, Children’s Hospital of Pittsburgh, Pittsburgh, PA, USA
†Department of Orthopedic Surgery, University of Pittsburgh, Pittsburgh, PA, USA
‡Department of Orthopedic Surgery, Kobe University Graduate School of Medicine, Kobe, Japan
§Departments of Molecular Genetics and Biochemistry, and Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA

We recently reported that the ruptured regions of the human anterior cruciate ligament (ACL) contained vascular-derived stem cells, which showed the potential for high expansion and multilineage differentiation. In this study, we performed experiments to test the hypothesis that ACL-derived CD34+ cells could contribute to tendon–bone healing. ACL-derived cells were isolated from the rupture site of human ACL by fluorescence-activated cell sorting. Following ACL reconstruction, immunodeficient rats received intracapsular administration of either ACL-derived CD34+ cells, nonsorted (NS) cells, CD34 cells, or phosphate-buffered saline (PBS). We also performed in vitro cell proliferation assays and enzyme-linked immunosorbent assays for vascular endothelial growth factor (VEGF) secretion. We confirmed the recruitment of the transplanted cells into the perigraft site after intracapuslar injection by immunohistochemical staining at week 1. Histological evaluation showed a greater area of collagen fiber formation and more collagen type II expression in the CD34+ group than the other groups at the week 2 time point. Immunostaining with isolectin B4 and rat osteocalcin demonstrated enhanced angiogenesis and osteogenesis in the CD34+ group at week 2. Moreover, double immunohistochemical staining for human-specific endothelial cell (EC) and osteoblast (OB) markers at week 2 demonstrated a greater ability of differentiation into ECs and OBs in the CD34+ group. Microcomputerized tomography showed the greatest healing of perigraft bone at week 4 in the CD34+ cell group, and the failure load of tensile test at week 8 demonstrated the greatest biomechanical strength in the CD34+ group. Furthermore, the in vitro studies indicated that the CD34+ group was superior to the other groups in their cell proliferation and VEGF secretion capacities. We demonstrated that ACL-derived CD34+ cells contributed to the tendon–bone healing after ACL reconstruction via the enhancement of angiogenesis and osteogenesis, which also contributed to an increase in biomechanical strength.

Key words: Anterior cruciate ligament; Stem cells; Reconstruction; Tendon–bone healing; Angiogenesis; Osteogenesis

Received December 17, 2010; final acceptance September 30, 2011. Online prepub date: June 20, 2012.
Address correspondence to Johnny Huard, Ph.D., Stem Cell Research Center, Children’s Hospital of Pittsburgh, Department of Orthopedic Surgery, University of Pittsburgh, 450 Technology Drive, 2 Bridgeside Point, Suite 206, Pittsburgh, PA 15219, USA. Tel: +1 412-648-2641; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1667–1677, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638937
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Endothelial Progenitor Cells Improve Directly and Indirectly Early Vascularization of Mesenchymal Stem Cell-Driven Bone Regeneration in a Critical Bone Defect in Rats

C. Seebach, D. Henrich, K. Wilhelm, J. H. Barker, and I. Marzi

Department of Trauma Surgery, Johann-Wolfgang-Goethe University, Frankfurt/Main, Germany

Early vascularization of a composite in a critical bone defect is a prerequisite for ingrowth of osteogenic reparative cells to regenerate bone, since lack of vessels does not ensure a sufficient nutritional support of the bone graft. The innovation of this study was to investigate the direct and indirect effects of endothelial progenitor cells (EPCs) and cotransplanted mesenchymal stem cells (MSCs) on the in vivo neovascularization activity in a critical size defect at the early phase of endochondral ossification. Cultivated human EPCs and MSCs were loaded onto β-TCP in vitro. A critical-sized bone defect (5 mm) was created surgically in the femoral diaphysis of adult athymic rat and stabilized with an external fixateur. The bone defects were filled with β-TCP, MSCs seeded on β-TCP, EPCs seeded on β-TCP, and coculture of MSCs and EPCs seeded on β-TCP or autologous bone of rat. After 1 week, the rats were sacrificed. Using quantitative CD34 immunohistochemistry as well as qualitative analysis of vascularization (staining of MHC and VEGF) in decalcified serial sections were performed by means of an image analysis system. Fluorescence microscopy analyzed the direct effects and indirect effects of human implanted EPCs for vessel formation at bone regeneration site. Formation of a primitive vascular plexus was also detectable in the β-TCP, MSC, or autologous bone group, but on a significantly higher level if EPCs alone or combined with MSCs were transplanted. Moreover, highest amount of vascularization were detected when EPCs and MSCs together were implanted. Early vascularization is improved by transplanted EPCs, which formed new vessels directly. Indeed the indirect effect of EPCs to vascularization is much higher. Transplanted EPC release chemotactic factors (VEGF) to recruit EPCs of the host and stimulate vascularization in the bone defect. Transplantation of human EPCs displays a promising approach to improve early vascularization of a scaffold in a critical bone defect. Moreover, coculture of EPCs and MSCs demonstrate also a synergistic effect on new vessel formation and seems to be a potential osteogenic construct for in vivo application.

Key words: Critical size defect; Coculture of EPCs and MSCs; Early vascularization; Bone regeneration; Cell transplantation

Received July 27, 2009; final acceptance October 21, 2011. Online prepub date: April 10, 2012.
Address correspondence to Caroline Seebach, M.D., Ph.D., Department of Trauma Surgery, Johann-Wolfgang-Goethe University Hospital, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany. Tel: +49(0)69-6301-7110; Fax: +49(0)69-6301-7108; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1679–1686, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X637434
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Mesenchymal Stromal Cell-Based Treatment of Jaw Osteoradionecrosis in Swine

Junji Xu,*1 Zongmei Zheng,*1 Dianji Fang,*1 Runtao Gao,* Yi Liu,* Zhipeng Fan,* Chunmei Zhang,* Songtao Shi,† and Songlin Wang*‡

*Molecular Laboratory for Gene Therapy and Tooth Regeneration, Capital Medical University School of Stomatology, Beijing, China
†Center for Craniofacial Molecular Biology, University of Southern California Ostrow, School of Dentistry, Los Angeles, CA, USA
‡Department of Biochemistry and Molecular Biology, Capital Medical University School of Basic Medical Sciences, Beijing, China

Jaw osteoradionecrosis (ORN) is a common and serious complication of radiation therapy for head and neck cancers. Bone marrow mesenchymal stromal cells (BMMSCs) are multipotent postnatal stem cells and have been widely used in clinical therapies. In the present study, we generated the mandibular ORN model in swine using a combination of single-dose 25-Gy irradiation and tooth extraction. A typical ORN phenotype, including loss of bone regeneration capacity and collagen collapse with the obliteration of vessels, gradually appeared after irradiation. After autologous BMMSC transplantation, new bone and vessels were regenerated, and the advanced mandibular ORN was treated successfully. In summary, we developed a swine model of jaw ORN, and our results indicate that autologous BMMSC transplantation may be a promising therapeutic approach for ORN.

Key words: Bone marrow mesenchymal stromal cells (BMMSCs); Jaw osteoradionecrosis (ORN); Bone regeneration; Large animal model; Swine

Received April 20, 2010; final acceptance October 28, 2011. Online prepub date: March 28, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Songlin Wang, Molecular Laboratory for Gene Therapy and Tooth Regeneration, Capital Medical University School of Stomatology, Tian Tan Xi Li No. 4, Beijing 100050, China. Tel: +86-10-67062012; Fax: +86-10-83911708; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1687–1696, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653039
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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In Vivo Differentiation of Human Amniotic Epithelial Cells Into Cardiomyocyte-Like Cells and Cell Transplantation Effect on Myocardial Infarction in Rats: Comparison With Cord Blood and Adipose Tissue-Derived Mesenchymal Stem Cells

Cheng-Hu Fang,*†1 Jiyong Jin,†1 Jun-Ho Joe,‡ Yi-Sun Song,‡ Byung-Im So,‡ Sang Moo Lim,§ Gi Jeong Cheon,§ Sang-Keun Woo,¶ Jeong-Chan Ra,# Young-Yiul Lee,** and Kyung-Soo Kim*

*Division of Cardiology, Hanyang University College of Medicine, Seoul, South Korea
†Division of Cardiology, Yanbian University, Yanji, China
‡Department of Biomedical Sciences, Hanyang University, Seoul, South Korea
§Department of Nuclear Medicine, Korea Cancer Center Hospital, Seoul, South Korea
¶Laboratory of Nuclear Medicine Research, Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul, South Korea
#Stem Cell Research Center, RNLBIO Co., Ltd., Seoul, South Korea
**Division of Hematology/Oncology, Hanyang University College of Medicine, Seoul, South Korea

Human amniotic epithelial cells (h-AECs), which have various merits as a cell source for cell therapy, are known to differentiate into cardiomyocytes in vitro. However, the ability of h-AECs to differentiate into cardiomyocytes in vivo and their cell transplantation effects on myocardial infarction are still unknown. In this study, we assessed whether h-AECs could differentiate into cardiomyocytes in vivo and whether h-AECs transplantation can decrease infarct size and improve cardiac function, in comparison to transplantation of cord blood-derived mesenchymal stem cells (MSCs) or adipose tissue-derived MSCs. For our study, we injected h-AECs, cord blood-derived MSCs, adipose tissue-derived MSCs, and saline into areas of myocardial infarction in athymic nude rats. After 4 weeks, 3% of the surviving h-AECs expressed myosin heavy chain, a marker specific to the myocardium. Compared with the saline group, all cell-implanted groups showed a higher ejection fraction, lower infarct area by positron emission tomography and histology, and more abundant myocardial gene and protein expression in the infarct area. We showed that h-AECs can differentiate into cardiomyocyte-like cells, decrease infarct size, and improve cardiac function in vivo. The beneficial effects of h-AECs were comparable to those of cord blood and adipose tissue-derived MSCs. These results support the need for further studies of h-AECs as a cell source for myocardial regeneration due to their plentiful availability, low immunity, and lack of ethical issues related to their use.

Key words: Human amniotic epithelial cells; Cardiomyocytes; Differentiation; Myocardial infarction

Received February 28, 2011; final acceptance October 10, 2011. Online prepub date: July 5, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Kyung-Soo Kim, M.D., Ph.D., Division of Cardiology, Department of Internal Medicine, Hanyang University College of Medicine, 17 Haengdang-dong, Sungdong-ku, Seoul, 133-792, South Korea. Tel: +82-2-2290-8312; Fax: +82-2-2298-9183; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1697–1709, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627462
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Intramyocardial Injections of Human Mesenchymal Stem Cells Following Acute Myocardial Infarction Modulate Scar Formation and Improve Left Ventricular Function

Jan Otto Beitnes,* Erik Øie,† Aboulghassem Shahdadfar,‡ Tommy Karlsen,‡# Regine M. B. Müller,‡ Svend Aakhus,* Finn P. Reinholt,§¶ and Jan E. Brinchmann‡#

*Department of Cardiology, Oslo University Hospital, Oslo, Norway
†Research Institute for Internal Medicine, Oslo University Hospital, Oslo, Norway
‡Institute of Immunology, Oslo University Hospital, Oslo, Norway
§Department of Pathology, Oslo University Hospital, Oslo, Norway
¶Institute of Pathology, University of Oslo, Oslo, Norway
#Norwegian Center for Stem Cell Research, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway

Cell therapy is a promising treatment modality to improve heart function in acute myocardial infarction. However, the mechanisms of action and the most suitable cell type have not been finally determined. We performed a study to compare the effects of mesenchymal stem cells (MSCs) harvested from different tissues on LV function and explore their effects on tissue structure by morphometry and histological staining for species and lineage relationship. MSCs from skeletal muscle (SM-MSCs) and adipose tissue (ADSCs) were injected in the myocardium of nude rats 1 week after myocardial infarction. After 4 weeks of observation, LVEF was significantly improved in the SM-MSCs group (39.1%) and in the ADSC group (39.6%), compared to the placebo group (31.0%, p < 0.001 for difference in change between groups). Infarct size was smaller after cell therapy (16.3% for SM-MSCs, 15.8% for ADSCs vs. 26.0% for placebo, p < 0.001), and the amount of highly vascularized granulation tissue in the border zone was significantly increased in both groups receiving MSCs (18.3% for SM-MSCs, 22.6% for ADSCs vs. 13.1% for placebo, p = 0.001). By in situ hybridization, moderate engraftment of transplanted cells was found, but no transdifferentiation to cardiomyocytes, endothelial cells, or smooth muscle cells was observed. We conclude that MSC injections lead to improved LVEF after AMI in rats predominantly by reduction of infarct size. After 4 weeks, we observed modulation of scar formation with significant increase in granulation tissue. Transdifferentiation of MSCs to cardiomyocytes or vascular cells did not contribute significantly in this process. MSCs from skeletal muscle and adipose tissue had similar effects.

Key words: Cell therapy; Mesenchymal stem cells (MSCs); In situ hybridization; Immunofluorescence; Echocardiography; Animal model

Received January 13, 2011; final acceptance June 25, 2011. Online prepub date: March 8, 2012.
Address correspondence to Jan E. Brinchmann, M.D., Ph.D., Norwegian Center for Stem Cell Research, Oslo University Hospital Rikshospitalet and University of Oslo, Domus Medica, Sognsvannsveien 9, PO Box 1121, Blindern, 0317 Oslo, Norway. Tel: +47 22 84 04 89; Fax: +47 22 85 10 58; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1711–1726, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640493
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
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Restricted Myogenic Potential of Mesenchymal Stromal Cells Isolated From Umbilical Cord

Iwona Grabowska,* Edyta Brzoska,* Agnieszka Gawrysiak,* Wladyslawa Streminska,* Jerzy Moraczewski,* Zbigniew Polanski,† Grazyna Hoser,‡ Jerzy Kawiak,‡ Eugeniusz K. Machaj,§¶ Zygmunt Pojda,§¶ and Maria A. Ciemerych*

*Department of Cytology, Institute of Zoology, Faculty of Biology, University of Warsaw, Warsaw, Poland
†Department of Genetics and Evolution, Institute of Zoology, Jagiellonian University, Cracow, Poland
‡Department of Clinical Cytology, Medical Centre of Postgraduate Education, Warsaw, Poland
§Department of Cellular Engineering, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland
¶Department of Regenerative Medicine, Military Institute of Hygiene and Epidemiology, Warsaw, Poland

Nonhematopoietic cord blood cells and mesenchymal cells of umbilical cord Wharton’s jelly have been shown to be able to differentiate into various cell types. Thus, as they are readily available and do not raise any ethical issues, these cells are considered to be a potential source of material that can be used in regenerative medicine. In our previous study, we tested the potential of whole mononucleated fraction of human umbilical cord blood cells and showed that they are able to participate in the regeneration of injured mouse skeletal muscle. In the current study, we focused at the umbilical cord mesenchymal stromal cells isolated from Wharton’s jelly. We documented that limited fraction of these cells express markers of pluripotent and myogenic cells. Moreover, they are able to undergo myogenic differentiation in vitro, as proved by coculture with C2C12 myoblasts. They also colonize injured skeletal muscle and, with low frequency, participate in the formation of new muscle fibers. Pretreatment of Wharton’s jelly mesenchymal stromal cells with SDF-1 has no impact on their incorporation into regenerating muscle fibers but significantly increased muscle mass. As a result, transplantation of mesenchymal stromal cells enhances the skeletal muscle regeneration.

Key words: Skeletal muscle regeneration; Stem cells; Human umbilical cord; Wharton’s jelly; Mouse

Received October 4, 2010; final acceptance October 10, 2011. Online prepub date: April 17, 2012.
Address correspondence to Maria A. Ciemerych, Department of Cytology, Institute of Zoology, Faculty of Biology, University of Warsaw, Ilji Miecznikowa 1, 02-096 Warsaw, Poland. Tel: +48 225542216; Fax: +48 225541203; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1727–1741, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X623925
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adipose Tissue-Derived Stem Cell Treatment Prevents Renal Disease Progression

Cassiano Donizetti-Oliveira,* Patricia Semedo,* Marina Burgos-Silva,* Marco Antonio Cenedeze,* Denise Maria Avancini Costa Malheiros,† Marlene A. Reis,‡ Alvaro Pacheco-Silva,* and Niels Olsen Saraiva Câmara*§

*Experimental and Clinical Immunology Laboratory, Division of Nephrology, Federal University of São Paulo, Escola Paulista de Medicina, São Paulo, Brazil
†Pathology Department, University of São Paulo, São Paulo, Brazil
‡Pathology Division, UFTM, Uberaba, Minas Gerais, Brazil
§Laboratory of Transplantation Immunobiology, Department of Immunology, University of São Paulo, São Paulo, Brazil

Adipose tissue-derived stem cells (ASCs) are an attractive source of stem cells with regenerative properties that are similar to those of bone marrow stem cells. Here, we analyze the role of ASCs in reducing the progression of kidney fibrosis. Progressive renal fibrosis was achieved by unilateral clamping of the renal pedicle in mice for 1 h; after that, the kidney was reperfused immediately. Four hours after the surgery, 2 × 105 ASCs were intraperitoneally administered, and mice were followed for 24 h posttreatment and then at some other time interval for the next 6 weeks. Also, animals were treated with 2 × 105 ASCs at 6 weeks after reperfusion and sacrificed 4 weeks later to study their effect when interstitial fibrosis is already present. At 24 h after reperfusion, ASC-treated animals showed reduced renal dysfunction and enhanced regenerative tubular processes. Renal mRNA expression of IL-6 and TNF was decreased in ASC-treated animals, whereas IL-4, IL-10, and HO-1 expression increased despite a lack of ASCs in the kidneys as determined by SRY analysis. As expected, untreated kidneys shrank at 6 weeks, whereas the kidneys of ASC-treated animals remained normal in size, showed less collagen deposition, and decreased staining for FSP-1, type I collagen, and Hypoxyprobe. The renal protection seen in ASC-treated animals was followed by reduced serum levels of TNF-α, KC, RANTES, and IL-1α. Surprisingly, treatment with ASCs at 6 weeks, when animals already showed installed fibrosis, demonstrated amelioration of functional parameters, with less tissue fibrosis observed and reduced mRNA expression of type I collagen and vimentin. ASC therapy can improve functional parameters and reduce progression of renal fibrosis at early and later times after injury, mostly due to early modulation of the inflammatory response and to less hypoxia, thereby reducing the epithelial–mesenchymal transition.

Key words: Adipose-derived stem cell; Acute kidney injury; Ischemia-reperfusion injury; Fibrosis; Inflammation

Received October 27, 2010; final acceptance September 20, 2011. Online prepub date: February 2, 2012.
Address correspondence to Niels Olsen Saraiva Câmara, M.D., Nephrology Division, Federal University of São Paulo, 740 Botucatu St., 04023-900, Vila Clementino, São Paulo, Brazil. Tel: (5511)-59041699; Fax: (5511)-59041894; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1743–1759, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627598
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Evaluation of Intracellular Labeling With Micron-Sized Particles of Iron Oxide (MPIOs) as a General Tool for In Vitro and In Vivo Tracking of Human Stem and Progenitor Cells

Jean-Luc Boulland,*†1 Doreen S. Y. Leung,*‡1 Marte Thuen,§1 Einar Vik-Mo,¶1 Mrinal Joel,*†¶ Marie-Claude Perreault,*† Iver A. Langmoen,†‡¶ Olav Haraldseth,§# and Joel C. Glover*†‡

*Department of Physiology, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway
†Norwegian Center for Stem Cell Research, Oslo University Hospital–National Hospital, Oslo, Norway
‡Center for Cancer Stem Cell Innovation, Oslo University Hospital–Radium Hospital, Oslo, Norway
§Department of Circulation and Medical Imaging, NTNU, Trondheim, Norway
¶Institute for Surgical Research and Department of Neurosurgery, Oslo University Hospital–National Hospital, Oslo, Norway
#Department of Radiology, St. Olav’s Hospital, Trondheim, Norway

Magnetic resonance imaging (MRI)-based tracking is increasingly attracting attention as a means of better understanding stem cell dynamics in vivo. Intracellular labeling with micrometer-sized particles of iron oxide (MPIOs) provides a practical MRI-based approach due to superior detectability relative to smaller iron oxide particles. However, insufficient information is available about the general utility across cell types and the effects on cell vitality of MPIO labeling of human stem cells. We labeled six human cell types from different sources: mesenchymal stem cells derived from bone marrow (MSCs), mesenchymal stem cells derived from adipose tissue (ASCs), presumptive adult neural stem cells (ad-NSCs), fetal neural progenitor cells (f-NPCs), a glioma cell line (U87), and glioblastoma tumor stem cells (GSCs), with two different sizes of MPIOs (0.9 and 2.84 μm). Labeling and uptake efficiencies were highly variable among cell types. Several parameters of general cell function were tested in vitro. Only minor differences were found between labeled and unlabeled cells with respect to proliferation rate, mitotic duration, random motility, and capacity for differentiation to specific phenotypes. In vivo behavior was tested in chicken embryos and severe combined immunodeficient (SCID) mice. Postmortem histology showed that labeled cells survived and could integrate into various tissues. MRI-based tracking over several weeks in the SCID mice showed that labeled GSCs and f-NPCs injected into the brain exhibited translocations similar to those seen for unlabeled cells and as expected from migratory behavior described in previous studies. The results support MPIO-based cell tracking as a generally useful tool for studies of human stem cell dynamics in vivo.

Key words: Adult stem cells; Progenitor cells; Neural stem cell; Mesenchymal stem cells; Stem cell transplantation; NOD/SCID chimeras; In vivo tracking; Cell migration

Received February 28, 2011; final acceptance October 21, 2011. Online prepub date: March 27, 2012.
1These authors provide equal contribution to this work.
Address correspondence to Joel C. Glover, Department of Physiology, Institute of Basic Medical Sciences, University of Oslo, PB 1103 Blindern, 0317 Oslo, Norway. Tel: +47-22851230; Fax: +47-22851249; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1761–1774, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X612521
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Antisense miR-7 Impairs Insulin Expression in Developing Pancreas and in Cultured Pancreatic Buds

Margarita Nieto,1 Pedro Hevia,1 Enrique Garcia,1 Dagmar Klein, Silvia Alvarez-Cubela, Valia Bravo-Egana, Samuel Rosero, R. Damaris Molano, Nancy Vargas, Camillo Ricordi, Antonello Pileggi, Juan Diez, Juan Domínguez-Bendala, and Ricardo L. Pastori

Diabetes Research Institute, University of Miami, Miller School of Medicine, Miami, FL, USA

MicroRNAs regulate gene expression by inhibiting translation or inducing target mRNA degradation. MicroRNAs regulate organ differentiation and embryonic development, including pancreatic specification and islet function. We showed previously that miR-7 is highly expressed in human pancreatic fetal and adult endocrine cells. Here we determined the expression profile of miR-7 in the mouse-developing pancreas by RT-PCR and in situ hybridization. MiR-7 expression was low between embryonic days e10.5 and e11.5, then began to increase at e13.5 through e14.5, and eventually decreased by e18. In situ hybridization and immunostaining analysis showed that miR-7 colocalizes with endocrine marker Isl1, suggesting that miR-7 is expressed preferentially in endocrine cells. Whole-mount in situ hybridization shows miR-7 highly expressed in the embryonic neural tube. To investigate the role of miR-7 in development of the mouse endocrine pancreas, antisense miR-7 morpholinos (MO) were delivered to the embryo at an early developmental stage (e10.5 days) via intrauterine fetal heart injection. Inhibition of miR-7 during early embryonic life results in an overall downregulation of insulin production, decreased b-cell numbers, and glucose intolerance in the postnatal period. This phenomenon is specific for miR-7 and possibly due to a systemic effect on pancreatic development. On the other hand, the in vitro inhibition of miR-7 in explanted pancreatic buds leads to b-cell death and generation of β-cells expressing less insulin than those in MO control. Therefore, in addition to the potential indirect effects on pancreatic differentiation derived from its systemic downregulation, the knockdown of miR-7 appears to have a β-cell-specific effect as well. These findings suggest that modulation of miR-7 expression could be utilized in the development of stem cell therapies to cure diabetes.

Key words: MicroRNA; b-Cells; Islets; Antisense microRNA; In vivo delivery

Received March 8, 2011; final acceptance June 1, 2011. Online prepub date: December 20, 2011.
1These authors contributed equally to this study.
Address correspondence to Ricardo L. Pastori, Ph.D., Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, 1450 NW 10th Avenue, Miami, FL 33136, USA. Tel: +1 (305) 243-5349; Fax: +1 (305) 243-4404; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it



Cell Transplantation, Vol. 21, pp. 1775–1789, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640628
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Molecularly Engineered Islet Cell Clusters for Diabetes Mellitus Treatment

Simmyung Yook,* Jee-Heon Jeong,* Yoon Suk Jung,* Sung Woo Hong,† Bok Hyeon Im,* Jin Won Seo,* Jun Beom Park,† Minhyung Lee,‡ Cheol-Hee Ahn,§ Haeshin Lee,¶ Dong Yun Lee,‡1 and Youngro Byun*†1

*College of Pharmacy, Seoul National University, Seoul, Republic of Korea
†WCU Departments of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, Republic of Korea
‡Department of Bioengineering, College of Engineering, and Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, Seoul, Republic of Korea
§Research Institute of Advanced Materials, Department of Materials Science and Engineering, Seoul National University, Seoul, Republic of Korea
¶Graduate School Nanoscience and Technology, College of Natural Science, Korea Advanced Institute of Science and Technology, Daejeon, South Korea

Pancreatic islet transplantation is a promising method for curing diabetes mellitus. We proposed in this study a molecularly engineered islet cell clusters (ICCs) that could overcome problems posed by islet transplantation circumstances and host’s immune reactions. A gene containing highly releasable exendin-4, an insulinotropic protein, was delivered into single islet cells to enhance glucose sensitivity; thereafter, the cells were reaggregated into small size ICCs. Then the surface of ICCs was modified with biocompatible poly(ethylene glycol)-lipid (PEG) (C18) for preventing immune reactions. The regimen of ICCs with low doses of anti-CD154 mAb and tacrolimus could effectively maintain the normal glucose level in diabetic mice. This molecularly engineered PEG-Sp-Ex-4 ICC regimen prevented cell death in transplantation site, partly through improving the regulation of glucose metabolism and by preventing hypoxia- and immune response-induced apoptosis. Application of this remedy is also potentially far-reaching; one would be to help overcome islet supply shortage due to the limited availability of pancreas donors and reduce the immunosuppressant regimens to eliminate their adverse effects.

Key words: Islet cell clusters (ICCs); Exendin-4; Polyethylene glycol; Transplantation

Received February 22, 2011; final acceptance August 25, 2011. Online prepub date: May 14, 2012.
1These authors provide equal contribution to this work.
Address correspondence to Youngro Byun, Ph.D., WCU Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, and College of Pharmacy, Seoul National University, Seoul 151-742, Republic of Korea. Tel: +82-2-880-9179; Fax: +82-2-872-7864; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Dong Yun Lee, Ph.D., Department of Bioengineering, College of Engineering, and Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, Seoul 133-791, Republic of Korea. Tel: +82-2-2220-2348; Fax: +82-2-2220-4741; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 1791–1802, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653011
E-ISSN 1555-3892
Copyright ©2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Early Islet Damage After Direct Exposure of Pig Islets to Blood: Has Humoral Immunity Been Underestimated?

Dirk J. van der Windt,*†1 Marco Marigliano,*‡1 Jing He,* Tatyana V. Votyakova,* Gabriel J. Echeverri,§¶ Burcin Ekser,§ David Ayares,# Fadi G. Lakkis,§** David K. C. Cooper,§ Massimo Trucco,* and Rita Bottino*

*Division of Immunogenetics, Department of Pediatrics, Children’s Hospital of Pittsburgh, Pittsburgh, PA, USA
†Department of Surgery, Erasmus University Medical Center, Rotterdam, The Netherlands
‡Regional Center for Diabetes in Children and Adolescents, Salesi’s Hospital, Ancona, Italy
§Thomas E. Starzl Transplantation Institute, Department of Surgery, University of Pittsburgh, Pittsburgh, PA, USA
¶Transplantation Unit, Fundacion Valle del Lili, Cali, Colombia
#Revivicor, Inc., Blacksburg, VA, USA
**Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA

Currently, islet transplantation as a cell therapeutic option for type 1 diabetes occurs via islet injection into the portal vein. Direct contact between islets and blood is a pathophysiological “provocation” that results in the instant blood-mediated inflammatory reaction (IBMIR) and is associated with early islet loss. However, the nature of the various insults on the islets in the blood stream remains mostly unknown. To gain insight into the mechanisms, we utilized a simplified in vitro model in which islets were exposed to blood in different clinically relevant but increasingly challenging, autologous, allogeneic, and xenogeneic combinations. Irrespective of the blood type and species compatibility, islets triggered blood clotting. Islet damage was worse as islet, and blood compatibility diminished, with substantial islet injury after exposure of porcine islets to human blood. Islet damage involved membrane leakage, antibody deposition, complement activation, positive staining for the membrane attack complex, and mitochondrial dysfunction. Islet damage occurred even after exposure to plasma only, and specific complement inactivation and neutralization of IgM substantially prevented islet damage, indicating the importance of humoral immunity. Efficacious measures are needed to reduce this injury, especially in view of a potential clinical use of porcine islets to treat diabetes.

Key words: Antibodies; Complement activation; Humoral immunity; IBMIR; Islet xenotransplantation

Received March 30, 2011; final acceptance October 5, 2011. Online prepub date: July 5, 2011.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Rita Bottino, Division of Immunogenetics, Rangos Research Center, 6th floor, Room 6126, Children’s Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA. Tel: +1 (412) 692-6644; Fax: +1 (412) 692-5809; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it