Cell Transplantation 25(12) Abstracts

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Cell Transplantation, Vol. 25, pp. 2083-2097, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X
692069
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Short-Term Grafting of Human Neural Stem Cells: Electrophysiological Properties and Motor Behavioral Amelioration in Experimental Parkinson’s Disease

Alberto Martínez-Serrano,*† Marta P. Pereira,*† Natalia Avaliani,‡ Anna Nelke,*† Merab Kokaia,‡ and Tania Ramos-Moreno*†‡

*Department of Molecular Biology, Universidad Autónoma de Madrid, Madrid, Spain
†Department of Molecular Neurobiology, Center of Molecular Biology Severo Ochoa (UAM-CSIC), Madrid, Spain
‡Epilepsy Center/Stem Cell Center, Wallenberg Neuroscience Center, Lund University Hospital, Lund, Sweden

Cell replacement therapy in Parkinson’s disease (PD) still lacks a study addressing the acquisition of electrophysiological properties of human grafted neural stem cells and their relation with the emergence of behavioral recovery after transplantation in the short term. Here we study the electrophysiological and biochemical profiles of two ventral mesencephalic human neural stem cell (NSC) clonal lines (C30-Bcl-XL and C32-Bcl-XL) that express high levels of Bcl-XL to enhance their neurogenic capacity, after grafting in an in vitro parkinsonian model. Electrophysiological recordings show that the majority of the cells derived from the transplants are not mature at 6 weeks after grafting, but 6.7% of the studied cells showed mature electrophysiological profiles. Nevertheless, parallel in vivo behavioral studies showed a significant motor improvement at 7 weeks postgrafting in the animals receiving C30-Bcl-XL, the cell line producing the highest amount of TH+ cells. Present results show that, at this postgrafting time point, behavioral amelioration highly correlates with the spatial dispersion of the TH+ grafted cells in the caudate putamen. The spatial dispersion, along with a high number of dopaminergic-derived cells, is crucial for behavioral improvements. Our findings have implications for long-term standardization of stem cell-based approaches in Parkinson’s disease.

Key words: Human neural stem cells (hNSCs); Parkinson’s disease (PD); A9-dopaminergic phenotype; Electrophysiology; Ventral mesencephalon

Received March 20, 2016; final acceptance September 13, 2016. Online prepub date: June 17, 2016.
Address correspondence to Tania Ramos-Moreno, Ph.D., Glioma Cell Therapy Group, Lund Stem Cell Center, Division of Neurosurgery/Department of Clinical Sciences, Lund University, BMC B10, SE-221 84 Lund, Sweden. Tel: +46 46 222 31 59; Fax: +46 46 2220560; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2099-2109, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691493
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

The Effects of Thermal Preconditioning on Oncogenic and Intraspinal Cord Growth Features of Human Glioma Cells

Xiang Zeng,*† Inbo Han,*†1 Muhammad Abd-El-Barr,* Zaid Aljuboori,*† Jamie E. Anderson,*† John H. Chi,* Ross D. Zafonte,‡ and Yang D. Teng*†‡

*Department of Neurosurgery, Harvard Medical School/Brigham and Women’s Hospital, Boston, MA, USA
†Division of SCI Research, Veterans Affairs Boston Healthcare System, Boston, MA, USA
‡Department of Physical Medicine and Rehabilitation, Harvard Medical School/Spaulding Rehabilitation Hospital, Boston, MA, USA

The adult rodent spinal cord presents an inhibitory environment for donor cell survival, impeding efficiency for xenograft-based modeling of gliomas. We postulated that mild thermal preconditioning may influence the fate of the implanted tumor cells. To test this hypothesis, high-grade human astrocytoma G55 and U87 cells were cultured under 37°C and 38.5°C to mimic regular experimental or core body temperatures of rodents, respectively. In vitro, the 38.5°C-conditioned cells, relative to 37°C, grew slightly faster. Compared to U87 cells, G55 cells demonstrated a greater response to the temperature difference. Hyperthermal culture markedly increased production of Hsp27 in most G55 cells, but only promoted transient expression of cancer stem cell marker CD133 in a small cell subpopulation. We subsequently transplanted G55 cells following 37°C or 38.5°C culture into the C2 or T10 spinal cord of adult female immunodeficient rats (3 rats/each locus/per temperature; total: 12 rats). Systematic analyses revealed that 38.5°C-preconditioned G55 cells grew more malignantly at either C2 or T10 as determined by tumor size, outgrowth profile, resistance to bolus intratumor administration of 5-fluorouracil (0.1 μmol), and posttumor survival (p < 0.05; n = 6/group). Therefore, thermal preconditioning of glioma cells may be an effective way to influence the in vitro and in vivo oncological contour of glioma cells. Future studies are needed for assessing the potential oncogenic modifying effect of hyperthermia regimens on glioma cells.

Key words: Spinal cord; Spinal cord glioma; Intramedullary tumor; Glioma; Glioblastoma; Tumor stem cell; Thermal stress; Preconditioning; Heat shock protein (HSP); Cell doubling time

Received January 5, 2016; final acceptance August 4, 2016. Online prepub date: May 4, 2016.
1Current affiliation: Department of Neurosurgery, CHA Bundang Medical Center, Seongnam, South Korea.
Address correspondence to Yang D. Teng, Harvard Medical School—Neurosurgery and PM&R, 221 Longwood Avenue, Boston, MA 02115, USA. Tel: 617-525-8676; Fax: 617-264-5216; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2111-2128, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692636
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Transplantation of Pro-Oligodendroblasts, Preconditioned by LPS-Stimulated Microglia, Promotes Recovery After Acute Contusive Spinal Cord Injury

Xiaojing Lin,*†‡ Tingbao Zhao,‡ Melissa Walker,† Aishi Ding,* Shide Lin,*‡ Yongcheng Cao,§ Jinfeng Zheng,§ Xiaohong Liu,§ Ming Geng,§ Xiao-Ming Xu,† and Shaojun Liu*

*Department of Neurobiology, Beijing Institute of Basic Medical Science, The Academy of Military Medical Sciences of the Chinese PLA, Beijing, P.R. China
†Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, and Department of Neurological Surgery, Indiana University School of Medicine, Indianapolis, IN, USA
‡Department of Spinal Cord Injury and Repair, Trauma and Orthopedics Institute of Chinese PLA, General Hospital of Jinan Military Region, Shandong Province, P.R. China
§Department of Pathology, General Hospital of Jinan Military Region, Shandong Province, P.R. China

Spinal cord injury (SCI) is a significant clinical challenge, and to date no effective treatment is available. Oligodendrocyte progenitor cell (OPC) transplantation has been a promising strategy for SCI repair. However, the poorposttransplantation survival and deficiency in differentiation into myelinating oligodendrocytes (OLs) are two major challenges that limit the use of OPCs as donor cells. Here we report the generation of an OL lineage population [i.e., pro-oligodendroblasts (proOLs)] that is relatively more mature than OPCs for transplantation after SCI. We found that proOLs responded to lipopolysaccharide (LPS)-stimulated microglia conditioned medium (L+M) by preserving toll-like receptor 4 (TLR4) expression, improving cell viability, and enhancing the expression of a myelinating OL marker myelin basic protein (MBP), compared to other OL lineage cells exposed to either LPS-stimulated (L+M) or nonstimulatedmicroglia conditioned medium (LM). When L+M-stimulated proOLs were intrathecally delivered through a lumbar puncture after a T10 thoracic contusive SCI, they promoted behavioral recovery, as assessed by the Basso–Beattie–Bresnahan (BBB) locomotor rating scale, stride length, and slips on the grid tests. Histologically, transplantation of L+proOLs caused a considerable increase in intralesional axon numbers and myelination, and less accumulation of invading macrophages when compared with the vehicle control or OPC transplantation. Thus, transplantation of proOLs, preconditioned by L+M, may offer a better therapeutic potential for SCI than OPCs since the former may have initiated the differentiation process toward OLs prior to transplantation.

Key words: Microglial activation; Oligodendrocyte progenitor cells (OPCs); Pro-oligodendroblasts (proOLs); Spinal cord injury (SCI); Transplantation

Received May 27, 2016; final acceptance September 14, 2016. Online prepub date: August 1, 2016.
Address correspondence to Xiao-Ming Xu, M.D., Ph.D., Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Indiana University School of Medicine, 320 West 15th Street, NB E500, Indianapolis, IN 46202-2266, USA. Tel: (317) 274-1036; Fax: (317) 278-5849; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Shaojun Liu, M.D., Ph.D., Department of Neurobiology, and Key Laboratory of Proteomics, Beijing Institute of Basic Medical Science, The Academy of Military Medical Sciences of the Chinese PLA, 27 Taiping Road, Beijing 100850, P.R. China. Tel: (86) 138-108-13837; Fax: (86) 10-68213039; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2129-2144, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692230
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Intravenously Infused F3.Olig2 Improves Memory Deficits via Restoring Myelination in the Aged Hippocampus Following Experimental Ischemic Stroke

Ji Hyeon Ahn,*1 Bai Hui Chen,†1 Bich Na Shin,‡ Jeong Hwi Cho,§ In Hye Kim,§ Joon Ha Park,* Jae Chul Lee,§ Hyun Jin Tae,¶ Yun Lyul Lee,† Jaesuk Lee,# Kyunghee Byun,#** Goo-Bo Jeong,** Bonghee Lee,#** Seung U. Kim,†† Young-Myeong Kim,‡‡ Moo-Ho Won,§ and Soo Young Choi*

*Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chuncheon, South Korea
†Department of Histology and Embryology, Institute of Neuroscience, Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China
‡Department of Physiology, College of Medicine, Institute of Neurodegeneration and NeuroregenerationHallym University, Chuncheon, South Korea
§Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, South Korea Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University, Iksan, South Korea
#Center for Genomics and Proteomics, Institute for Regenerative Medicine, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon, South Korea
**Department of Anatomy and Cell Biology, Gachon University Graduate School of Medicine, Incheon, South Korea
††Department of Medicine, University of British Columbia, Vancouver, British Columbia, Canada
‡‡Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon, South Korea

Oligodendrocytes play a crucial role in creating the myelin sheath that is an important component in neural transmission. In an animal model of transient cerebral ischemia, application of oligodendrocyte progenitor cells (OPCs) has not yet been reported. In this study, the effects of F3.Olig2 transplantation on memory and cognitive dysfunction were investigated in the aged gerbil in which ischemic stroke was induced. To investigate the possible mechanisms underlying repair, changes in the expression of myelin basic protein (MBP), oligodendrocyte-specific protein (OSP), and brain-derived neurotrophic factor (BDNF) were examined. Experimental ischemic stroke was induced by occlusion of bilateral common carotid arteries in aged gerbils. Gerbils (n = 31 per group) were randomly divided into three groups: (1) vehicle sham group, (2) vehicle ischemia group, and (3) F3.Olig2 ischemia group. After 1, 3, and 7 days of ischemia–reperfusion (I-R), saline or F3.Olig2 cells (1 × 106 cells in 100 μl) were injected into the gerbils intravenously. The gerbils were sacrificed 10 days after I-R for identification of grafted F3.Olig2 cells, and 15 and 30 days after I-R for tissue analysis after conducting passive avoidance and novel object recognition test. Injected F3.Olig2 cells and MBP, OSP, and BDNF were detected by specific antibodies using immunohistochemistry and/or Western blots. Memory and cognition were significantly increased in the F3.Olig2 ischemia group compared with the vehicle ischemia group. In the F3.Olig2 ischemia group, the neurons were not protected from ischemic damage; however, MBP, OSP, and BDNF expressions were significantly increased. Our results show that injection of F3.Olig2 cells significantly improved impaired memory and cognition, which might be related to increased MBP expression via increasing OSP and BDNF expression in the aged gerbil hippocampus following transient cerebral ischemia.

Key words: Brain-derived neurotrophic factor (BDNF); Cognitive impairment; Ischemia–reperfusion injury; Neural stem cells (NSCs); Olig2 cDNA; Oligodendrocyte-specific protein (OSP)

Received January 25, 2016; final acceptance September 13, 2016. Online prepub date: July 20, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Professor Moo-Ho Won, DVM, Ph.D., Department of Neurobiology, School of Medicine, Kangwon National University, 1 Kangwondaehak-gilChuncheonGangwon 24341, South Korea. Tel: +82-33-250-8891; Fax: +82-33-256-1614; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Professor Soo Young Choi, Ph.D., Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, 1 Hallymdaehak-gilChuncheonGangwon 24252, South Korea. Tel: +82-33-248-2112; Fax: +82-33-241-1463; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2145-2156, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692005
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Automated Analysis of Microscopic Images of Isolated Pancreatic Islets

David Habart,* Jan Švihlík,†‡ Jan Schier,§ Monika Cahová,¶ Peter Girman,* Klára Zacharovová,¶ Zuzana Berková,¶ Jan Kříž,* Eva Fabryová,¶ Lucie Kosinová,¶ Zuzana Papáčková,¶ Jan Kybic,† and František Saudek*

*Diabetes Center, Institute for Clinical and Experimental Medicine (IKEM), Prague, Czech Republic
†Biomedical Imaging Algorithms Group, Department of Cybernetics, Faculty of Electrical Engineering, Czech Technical University in Prague, Prague, Czech Republic
‡University of Chemistry and Technology, Prague, Czech Republic
§Department of Image Processing, Institute of Information Theory and Automation, The Czech Academy of Sciences, Prague, Czech Republic
¶Center of Experimental Medicine, Institute for Clinical and Experimental Medicine (IKEM), Prague, Czech Republic

Clinical islet transplantation programs rely on the capacities of individual centers to quantify isolated islets. Current computer-assisted methods require input from human operators. Here we describe two machine learning algorithms for islet quantification: the trainable islet algorithm (TIA) and the nontrainable purity algorithm (NPA). These algorithms automatically segment pancreatic islets and exocrine tissue on microscopic images in order to count individual islets and calculate islet volume and purity. References for islet counts and volumes were generated by the fully manual segmentation (FMS) method, which was validated against the internal DNA standard. References for islet purity were generated via the expert visual assessment (EVA) method, which was validated against the FMS method. The TIA is intended to automatically evaluate micrographs of isolated islets from future donors after being trained on micrographs from a limited number of past donors. Its training ability was first evaluated on 46 images from four donors. The pixel-to-pixel comparison, binary statistics, and islet DNA concentration indicated that the TIA was successfully trained, regardless of the color differences of the original images. Next, the TIA trained on the four donors was validated on an additional 36 images from nine independent donors. The TIA was fast (67 s/image), correlated very well with the FMS method (R2= 1.00 and 0.92 for islet volume and islet count, respectively), and had small REs (0.06 and 0.07 for islet volume and islet count, respectively). Validation of the NPA against the EVA method using 70 images from 12 donors revealed that the NPA had a reasonable speed (69 s/image), had an acceptable RE (0.14), and correlated well with the EVA method (R2 = 0.88). Our results demonstrate that a fully automated analysis of clinical-grade micrographs of isolated pancreatic islets is feasible. The algorithms described herein will be freely available as a Fiji platform plugin.

Key words: Islet transplantation; Enumeration of islets; Quality control; Image processing; Image segmentation; Machine learning

Received February 20, 2015; final acceptance September 20, 2016. Online prepub date: June 9, 2016.
Address correspondence to David Habart, M.D., Ph.D., Diabetes Centre, Institute for Clinical and Experimental Medicine, Vídeňská 1958/9, PSČ 140 21, Prague 4, Czech Republic. Tel: +420 262 362 288; Fax: +420 261 362 820; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2157-2171, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691132
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Intravenous Injection of Clinical Grade Human MSCs After Experimental Stroke: Functional Benefit and Microvascular Effect

Anaïck Moisan,*†‡ Isabelle Favre,*†§ Claire Rome,*† Florence De Fraipont,¶# Emmanuelle Grillon,*† Nicolas Coquery,*† Hervé Mathieu,*† Virginie Mayan,*† Bernadette Naegele,*†§ Marc Hommel,** Marie-Jeanne Richard,‡¶# Emmanuel Luc Barbier,*† Chantal Remy,*† and Olivier Detante*†§

*The French National Institute of Health and Medical Research (Inserm), Grenoble, France
Université Grenoble Alpes, Grenoble, France
‡French Blood Company/CHU de Grenoble, Hôpital MichallonUnité de Thérapie et d’Ingénierie Cellulaire, Saint Ismier, France
§Unité NeurovasculaireDépartement de NeurologieHôpital Michallon, CHU Grenoble Alpes, Grenoble, France
¶UM Biochimie des Cancers et BiothérapiesHôpital Michallon, CHU Grenoble Alpes, Grenoble, France
#Institut Albert BonniotInsermUniversité Grenoble Alpes, Grenoble, France
**Département de Recherche Clinique, Hôpital Michallon, CHU de Grenoble, Grenoble, France

Stroke is the leading cause of disability in adults. Many current clinical trials use intravenous (IV) administration of human bone marrow-derived mesenchymal stem cells (BM-MSCs). This autologous graft requires a delay for ex vivo expansion of cells. We followed microvascular effects and mechanisms of action involved after an IV injection of human BM-MSCs (hBM-MSCs) at a subacute phase of stroke. Rats underwent a transient middle cerebral artery occlusion (MCAo) or a surgery without occlusion (sham) at day 0 (D0). At D8, rats received an IV injection of 3 million hBM-MSCs or PBS-glutamine. In a longitudinal behavioral follow-up, we showed delayed somatosensory and cognitive benefits 4 to 7 weeks after hBM-MSC injection. In a separate longitudinal in vivo magnetic resonance imaging (MRI) study, we observed an enhanced vascular density in the ischemic area 2 and 3 weeks after hBM-MSC injection. Histology and quantitative polymerase chain reaction (qPCR) revealed an overexpression of angiogenic factors such as Ang1 and transforming growth factor-β1 (TGF-β1) at D16 in hBM-MSC-treated MCAo rats compared to PBS-treated MCAo rats. Altogether, delayed IV injection of hBM-MSCs provides functional benefits and increases cerebral angiogenesis in the stroke lesion via a release of endogenous angiogenic factors enhancing the stabilization of newborn vessels. Enhanced angiogenesis could therefore be a means of improving functional recovery after stroke.

Key words: Mesenchymal stem cells (MSCs); Cell therapy; Cerebral ischemia; Stroke; Microvasculature; Angiogenesis

Received December 21, 2014; final acceptance May 20, 2016. Online prepub date: February 26, 2016.
Address correspondence to Anaïck MoisanUnité de Thérapie et d’Ingénierie Cellulaire–EFS Rhône Alpes, 464 route de Lancey, 38330 Saint Ismier, France. Tel: 0033. (0)4.76.52.92.41; Fax: 0033. (0)4.76.52.59.84; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2173-2185, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691132
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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The Dose of Intravenously Transplanted Bone Marrow Stromal Cells Determines the Therapeutic Effect on Vascular Remodeling in a Rat Model of Ischemic Stroke

Bin He,*1 Qianmin Yao,*1 Zijing Liang,* Junmin Lin,* Yubao Xie,* Shaoming Li,* Gongxiong Wu,† Zhihua Yang,* and Pingyi Xu*

*Department of Neurology, The First Affiliated Hospital of Guangzhou Medical University, Guangdong, P.R. China
†Cardiovascular Center, The Second Affiliated Hospital of Guangzhou Medical University, Guangdong, P.R. China

The therapeutic benefits of bone marrow-derived mesenchymal stem cell (BM-MSC) transplantation for ischemic stroke have been extensively demonstrated. However, studies on the optimal cell dose for intravenous administration are still limited. This study aimed to determine an appropriate cell dose for BM-MSC intravenous transplantation and to investigate the effect of cell dose on vascular remodeling in a rat model of ischemic stroke. BM-MSCs at doses of 5 × 104(low-dose group), 5 × 105 (medium-dose group), and 2 × 106 (high-dose group) were intravenously injected into rats at 72 h after ischemia. The therapeutic efficacy of BM-MSCs was evaluated by measuring infarct volume, vascular diameters, capillary area in the peri-infarct zone, level of basic fibroblast growth factor (bFGF) in the peri-infarct zone, and serum vascular endothelial growth factor (VEGF) level at 7 days after ischemia. Compared with the low-dose and control groups, medium-dose and high-dose BM-MSC transplantation significantly reduced the volume of the infarct area, enlarged the diameters of pial vessels and the basilar artery, and increased the capillary area in the peri-infarct zone of the cerebral cortex. Furthermore, transplanted BM-MSCs elevated the expressions of bFGF in the peri-infarct zone and the serum VEGF level. Administration of 5 × 105 BM-MSCs is an appropriate cell dose for ischemic stroke therapy in rats. These findings may be helpful for designing future clinical trials.

Key words: Cerebral infarction; Bone marrow-derived mesenchymal stem cells (BM-MSCs); Transplantation; Vascular endothelial growth factor (VEGF); Basic fibroblast growth factor (bFGF)

Received April 22, 2016; final acceptance September 14, 2016. Online prepub date: August 1, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Zhihua Yang, Department of Neurology, The First Affiliated Hospital of Guangzhou Medical University, No. 151 YanJiang Road, YueXiu District, Guangzhou 510120, P.R. China. Tel: +86-15602288528; Fax: +86-83062767; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Pingyi Xu, No. 151 YanJiang Road, YueXiu District, Guangzhou 510120, P.R. China. Tel: +86-13826161149; Fax: +86-83062767; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2187-2197, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692753
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Bone Marrow-Derived Endothelial Progenitor Cells Reduce Recurrent Miscarriage in Gestation

Kazuyoshi Kanki,* Masaaki Ii,†‡ Yoshito Terai,* Masahide Ohmichi,* and Michio Asahi†

*Department of Gynecology, Faculty of Medicine, Osaka Medical College, Osaka, Japan
†Department of Pharmacology, Faculty of Medicine, Osaka Medical College, Osaka, Japan
‡Division of Research Animal Laboratory and Translational Medicine, Research and Development Center, Osaka Medical College, Osaka, Japan

Bone marrow-derived endothelial progenitor cells (EPCs) have been shown to contribute to not only angiogenesis in ischemic tissue but also neovascularization in uterine endometrium formation. Reduced neovascularization and elevation of serum soluble Flt1, a functional blockage of VEGF, in the development of placenta is thought to be one of the major causes of repeated miscarriages in gestation. We then examined whether transfusion of VEGF-expressing extrinsic EPCs prevented frequent miscarriage via its promotional effect on neovascularization with a VEGF–eNOS signaling pathway in a mouse miscarriage model. The results showed that systemic EPC transfusion significantly reduced the rate of miscarriage, and EPCs were frequently observed in the miscarriage placenta. In contrast, only a few EPCs were detected in the placenta of normal gestation. The vascular pattern was irregular, and vessel size was small in the miscarriage placenta compared with that of normal gestation. The placental vascular pattern in miscarriage tended to be normalized with increased vessel size up to a similar level as normal gestation by EPC recruitment. For the mechanistic insight, since soluble Flt1 inhibits EPC functions, it was suggested that the increased soluble Flt1 could suppress the recruited EPC functional activity in the miscarriage placenta. In vitro experiments by soluble Flt1 treatment in cultured EPCs suggested that the vascular abnormality could be partly due to the inhibition of eNOS expression by the increased amounts of soluble Flt1. These findings from animal experiments indicated that autologous EPC therapy may be a novel therapy to prevent miscarriage in high-risk pregnancies, such as preeclampsia.

Key words: Bone marrow-derived stem cells (BMSCs); Endothelial progenitor cells (EPCs); Miscarriage; Vascular development; Placenta

Received October 7, 2015; final acceptance August 31, 2016. Online prepub date: August 5, 2016.
Address correspondence to Masaaki Ii, M.D., Ph.D., Division of Research Animal Laboratory and Translational Medicine, Research and Development Center, Osaka Medical College, 2-7 Daigaku-machi, Takatsuki, Osaka 569-8686, Japan. Tel: +81-72-684-6537; Fax: +81-72-684-7042; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2187-2197, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692104
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Long-Term Cell Tracking Following Local Injection of Mesenchymal Stromal Cells in the Equine Model of Induced Tendon Disease

Janina Burk,*†‡ Dagmar Berner,§ Walter Brehm,*†§ Aline Hillmann,*† Carolin Horstmeier,*†§ Christoph Josten,¶ Felicitas Paebst,§ Giacomo Rossi,# Susanna Schubert,*† and Annette B. Ahrberg*†¶

*Saxon Incubator for Clinical Translation (SIKT), University of Leipzig, Leipzig, Germany
†Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig, Germany
‡Institute of Veterinary Physiology, University of Leipzig, Leipzig, Germany
§Large Animal Clinic for Surgery, University of Leipzig, Leipzig, Germany
¶Department of Orthopedics, Traumatology and Plastic Surgery, University of Leipzig, Leipzig, Germany
#University of Camerino, School of Biosciences and Veterinary Medicine, Matelica (MC), Italy

Tendon disease has been treated with multipotent mesenchymal stromal cells (MSCs) in the equine large-animal model with promising success. The aim of this study was to gain more insight into the fate and biodistribution of MSCs after local application into tendon lesions by long-term cell tracking in this large-animal model. Superficial digital flexor tendon lesions were induced in all limbs in six horses and injected with 10 × 106 Molday ION Rhodamine B™-labeled MSCs suspended in serum or serum alone. Follow-up was performed using low-field magnetic resonance imaging (MRI), flow cytometry, and histology. Cell tracking based on the hypointense artifacts induced by the superparamagnetic iron oxide (SPIO) labeling agent in MRI as well as based on Rhodamine B fluorescence was feasible. However, Prussian blue staining for assessment of histology was not entirely specific for SPIO. Labeled cells could be traced at their injection site by MRI as well as histology for the whole follow-up period of 24 weeks. Although the numbers of labeled cells within the injected tendon lesions decreased over time, part of the applied cells appeared to remain viable and integrated within the injured tissue. Furthermore, small numbers of labeled cells were identified in peripheral blood within the first 24 h after cell injection and could also be found until week 24 within the contralateral control tendon lesions that had been injected with serum. The present findings unveil details on MSC biodistribution and persistence after their local application, which are of clinical relevance with regard to MSC safety and mechanisms of action.

Key words: Tendon; Mesenchymal stromal cells (MSCs); Cell therapy; Cell tracking; Biodistribution; Superparamagnetic iron oxide (SPIO)

Received December 23, 2015; final acceptance September 7, 2016. Online prepub date: July 7, 2016.
Address correspondence to Dr. Janina Burk, Saxon Incubator of Clinical Translation (SIKT), University of Leipzig, Philipp-Rosenthal-Str. 55, 04103 Leipzig, Germany. Tel: +49-341-9739616; Fax: +49-341-9738269; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2213-2220, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692618
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Stem Cell Delivery With Polymer Hydrogel for Treatment of Intervertebral Disc Degeneration: From 3D Culture to Design of the Delivery Device for Minimally Invasive Therapy

Deepak Kumar,*1 Alex Lyness,* Irini Gerges,†2 Christina Lenardi,†3 Nicholas R. Forsyth,‡ and Yang Liu*

*Centre of Biological Engineering, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough, UK
†Fondazione FilareteViale Ortles, Milan, Italy
‡Guy Hilton Research Centre, Keele University, Staffordshire, UK

Nucleus pulposus (NP) tissue damage can induce detrimental mechanical strain on the biomechanical performance of intervertebral discs (IVDs), causing subsequent disc degeneration. A novel, photocurable, injectable, synthetic polymer hydrogel (pHEMA-co-APMA grafted with PAA) has already demonstrated success in encapsulating and differentiating human mesenchymal stem cells (hMSCs) toward an NP phenotype during hypoxic conditions. After demonstration of promising results in our previous work, in this study we have further investigated the inclusion of mechanical stimulation and its impact on hMSC differentiation toward an NP phenotype through the characterization of matrix markers such as SOX-9, aggrecan, and collagen II. Furthermore, investigations were undertaken in order to approximate delivery parameters for an injection delivery device, which could be used to transport hMSCs suspended in hydrogel into the IVD. hMSC-laden hydrogel solutions were injected through various needle gauge sizes in order to determine its impact on postinjection cell viability and IVD tissue penetration. Interpretation of these data informed the design of a potential minimally invasive injection device, which could successfully inject hMSCs encapsulated in a UV-curable polymer into NP, prior to photo-cross-linking in situ.

Key words: Regenerative medicine; Cell encapsulation; Tissue engineering; Cell delivery; Device design

Received August 27, 2016; final acceptance September 22, 2016. Online prepub date: July 22, 2016.
1Current affiliation: School of Materials, University of Manchester, Manchester, UK.
2Current affiliation: Tensive s.r.l., Via Timavo, Milan, Italy.
3Current affiliation: CIMAINA, Dipartimento di FisicaUniversità degli Studi di Milano, Via Celoria, Milan, Italy.
Address correspondence to Yang Liu, Centre of Biological Engineering, Wolfson School of Mechanical, Electrical and Manufacturing Engineering, Loughborough University, Loughborough, Leicestershire LE11 3TU, UK. Tel: +44 (0) 1509 227602; Fax: +44 (0) 1509 227648; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2221-2243, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692014
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Human Urinary Epithelial Cells as a Source of Engraftable Hepatocyte-Like Cells Using Stem Cell Technology

Vanessa Sauer,*† Tatyana Tchaikovskaya,*† Xia Wang,*† Yanfeng Li,*† Wei Zhang,†‡§ Krisztina Tar,*†1 Zsuzsanna Polgar,*† Jianqiang Ding,*† Chandan Guha,†‡§ Ira J. Fox,¶ Namita Roy-Chowdhury,*†# and Jayanta Roy-Chowdhury*†#

*Department of Medicine, Albert Einstein College of Medicine, New York, NY, USA
†Marion Bessin Liver Research Center, Albert Einstein College of Medicine, New York, NY, USA
‡Department of Radiation Oncology, Albert Einstein College of Medicine, New York, NY, USA
§Department of Pathology, Albert Einstein College of Medicine, NY, New York, USA
¶Department of Surgery and McGowan Institute for Regenerative Medicine, Children’s Hospital of Pittsburgh of University of Pittsburgh Medical Center, Pittsburgh, PA, USA
#Department of Genetics, Albert Einstein College of Medicine, New York, NY, USA

Although several types of somatic cells have been reprogrammed into induced pluripotent stem cells (iPSCs) and then differentiated to hepatocyte-like cells (iHeps), the method for generating such cells from renal tubular epithelial cells shed in human urine and transplanting them into animal livers has not been described systematically. We report reprogramming of human urinary epithelial cells into iPSCs and subsequent hepatic differentiation, followed by a detailed characterization of the newly generated iHeps. The epithelial cells were reprogrammed into iPSCs by delivering the pluripotency factors OCT3/4, SOX2, KLF4, and MYC using methods that do not involve transgene integration, such as nucleofection of episomal (oriP/EBNA-1) plasmids or infection with recombinant Sendai viruses. After characterization of stable iPSC lines, a three-step differentiation toward hepatocytes was performed. The iHeps expressed a large number of hepatocyte-preferred genes, including nuclear receptors that regulate genes involved in cholesterol homeostasis, bile acid transport, and detoxification. MicroRNA profile of the iHeps largely paralleled that of primary human hepatocytes. The iHeps engrafted into the livers of Scid mice transgenic for mutant human SERPINA1 after intrasplenic injection. Thus, urine is a readily available source for generating human iHeps that could be potentially useful for disease modeling, pharmacological development, and regenerative medicine.

Key words: Urinary epithelial cells; Induced pluripotent stem cells (iPSCs); Differentiation; Hepatocytes

Received August 21, 2015; final acceptance September 7, 2016. Online prepub date: June 9, 2016.
1Current affiliation: University of Debrecen, Faculty of Medicine, Department of Medical Chemistry, Debrecen, Hungary.
Address correspondence to Vanessa Sauer at her current affiliation: Klinik für TransplantationsmedizinUniversitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A 14, 48145 Münster, Germany. Tel: +49-251-83-57935; Fax: +49-251-83-57949; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Jayanta Roy-Chowdhury, Albert Einstein College of Medicine, Jack and Pearl Resnick Campus, 1300 Morris Park Avenue, Ullmann Building, Room 523, Bronx, NY 10461, USA. Tel: +1-718-430-2265; Fax: +1-718-430-8975; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2245-2257, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692645
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Antifibrotic Effects of Human Amniotic Membrane Transplantation in Established Biliary Fibrosis Induced in Rats

Luciana B. Sant’Anna,* Raduan Hage,* Maria Angélica G. Cardoso,* Emilia A. L. Arisawa,*

Maria Martin Cruz,* Ornella Parolini,† Anna Cargnoni,† and Nilson Sant’Anna

*Institute of Research and Development, University of Vale do Paraíba (UNIVAP), São José dos Campos, São Paulo, Brazil
†Centro di Ricerca E. Menni (CREM), Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy
‡Computing and Applied Mathematics Laboratory, National Institute for Space Research (INPE), São José dos Campos, São Paulo, Brazil

Liver fibrosis is characterized by excessive accumulation of extracellular matrix components in the liver parenchyma that distorts the normal architecture and hepatic function. Progressive fibrosis could end in the advanced stage known as cirrhosis, resulting in the need to resort to liver transplantation. Amniotic membrane (AM) has emerged as an innovative therapeutic approach for chronic liver diseases due to its anti-inflammatory, antiscarring, and wound-healing effects. We have recently shown that AM can be used as a patch on the liver surface at the same time of fibrosis induction, resulting in significantly reduced progression and severity of biliary fibrosis. Here we investigated the effects of human AM on the established rat model of liver fibrosis, induced by the bile duct ligation (BDL). We also explored the effect of AM on the expression of transforming growth factor-β1 (TGF-β1), the main profibrogenic factor in hepatic fibrosis, and the proinflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and anti-inflammatory cytokine IL-10. Two weeks after BDL, the liver was covered with a fragment of AM or left untreated. Six weeks later, the fibrosis was first assessed by the semiquantitative Knodell and the METAVIR scoring systems and, thereafter, by CellProfiler digital image analysis to quantify the area occupied by collagen deposition, ductular reactions (DRs), activated myofibroblasts, and TGF-β1. The hepatic cytokines were determined by ELISA. AM-treated rats showed a significantly lower score compared to the control BDL rats (2.5 ± 0.9 vs. 3.5 ± 0.3, respectively; p < 0.05). The collagen deposition, DRs, number of activated myofibroblasts, and TGF-β1 were all reduced to about 50% of levels observed in untreated BDL rats. These findings suggest that AM, when applied as a patch onto the liver surface, is useful for treating well-established cholestatic fibrosis, and the mechanism was partly by means of downregulating the profibrotic factor TGF-β1 and IL-6.

Key words: Amniotic membrane (AM); Bile duct ligation (BDL); Digital image analysis; Immunohistochemistry; ELISA

Received June 11, 2015; final acceptance September 13, 2016. Online prepub date: August 1, 2016.
Address correspondence to Luciana Barros Sant’AnnaInstituto de PesquisaDesenvolvimentoUniversidade do Vale do Paraíba (UNIVAP), Av. Shishima Hifumi, 2911, Urbanova, CEP: 12244000, São José dos Campos, São Paulo, Brazil. Tel: 55-12-39471169; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2259-2268, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692023
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Alloimmune Monitoring After Islet Transplantation: A Prospective Multicenter Assessment of 25 Recipients

Vaihere Delaune,*† Christian Toso,*† Pierre-Yves Benhamou,‡ Anne Wojtusciszyn,§ Laurence Kessler,¶ Florence Slits,* Sandrine Demuylder-Mischler,† Nadine Pernin,# Sandrine Lablanche,‡ Lorenzo A. Orci,*† Graziano Oldani,*† Philippe Morel,† Thierry Berney,†# and Stéphanie Lacotte*

*Hepatology and Transplantation Laboratory, Department of Surgery, Faculty of Medicine, University of Geneva, Geneva, Switzerland
†Divisions of Abdominal and Transplantation Surgery, Department of Surgery, Geneva University Hospitals, Geneva, Switzerland
‡Division of Endocrinology, Grenoble University Hospital, Grenoble, France
§Division of Endocrinology, Diabetes and Nutrition, and Laboratory for Diabetes Cell Therapy, Montpellier University Hospital, Montpellier, France
¶Division of Diabetology, Strasbourg University Hospital, Strasbourg, France
#Cell Transplantation Laboratory, Department of Surgery, Faculty of Medicine, University of Geneva, Geneva, Switzerland

Islet transplantation is an effective treatment for selected patients with type 1 diabetes. However, an accurate test still lacks for the early detection of graft rejection. Blood samples were prospectively collected in four university centers (Geneva, Grenoble, Montpellier, and Strasbourg). Peripheral blood mononuclear cells were stimulated with donor splenocytes in the presence of interleukin-2. After 24 h of incubation, interferon-γ (IFN-γ) ELISpot analysis was performed. After a total of 5 days of incubation, cell proliferation was assessed by fluorescence-activated cell sorting (FACS) analysis for Ki-67. Immunological events were correlated with adverse metabolic events determined by loss of ≥1 point of β-score and/or an increased insulin intake ≥10%. Twenty-five patients were analyzed; 14 were recipients of islets alone, and 11 combined with kidney. Overall, 76% (19/25) reached insulin independence at one point during a mean follow-up of 30.7 months. IFN-g ELISpot showed no detectable correlation with adverse metabolic events [area under the curve (AUC) = 0.57]. Similarly, cell proliferation analysis showed no detectable correlation with adverse metabolic events (CD3+/CD4+AUC = 0.54; CD3+/CD8+ AUC = 0.55; CD3/CD56+ AUC = 0.50). CD3/CD56+ cell proliferation was significantly higher in patients with combined kidney transplantation versus islet alone (6 months, p = 0.010; 12 months, p = 0.016; and 24 months, p = 0.018). Donor antigen-stimulated IFN-γ production and cell proliferation do not predict adverse metabolic events after islet transplantation. This suggests that the volume of transplanted islets is too small to produce a detectable systemic immune response and/or that alloimmune rejection is not the sole reason for the loss of islet graft function.

Key words: Diabetes; Islet transplantation; Alloimmune monitoring; ELISpot; Cell proliferation

Received April 7, 2016; final acceptance August 31, 2016. Online prepub date: June 13, 2016.
Address correspondence to Stéphanie Lacotte, Transplantation Laboratory, Medical University Center, Rue Michel Servet 1, 1205 Geneva, Switzerland. Tel: +41 22 379 56 61; Fax: +41 22 379 59 55; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2269-2276, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691709
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Evaluation of Perfluorohexyloctane/Polydimethylsiloxane for Pancreas Preservation for Clinical Islet Isolation and Transplantation

Magnus Ståhle,* Aksel Foss,† Bengt Gustafsson,‡ Marko Lempinen,§ Torbjörn Lundgren,¶ Ehab Rafael,# Gunnar Tufveson,** Bastian Theisinger,†† Daniel Brandhorst,* Olle Korsgren,* and Andrew Friberg*

*Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
†Department of Transplantation Surgery, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway
‡Department of Transplantation, University Hospital, Gothenburg, Sweden
§Division of Transplantation, Surgical Hospital, Helsinki University, Helsinki, Finland
¶Division of Transplantation Surgery, CLINTEC, Karolinska Institute, Stockholm, Sweden
#Department of Nephrology and Transplantation, University Hospital, Malmö, Sweden
**Department of Surgical Sciences, Division of Transplantation Surgery, Uppsala University Hospital, Uppsala, Sweden
††Novaliq GmbH, Heidelberg, Germany

This study aimed to evaluate a 50:50 mix of perfluorohexyloctane/polydimethylsiloxane 5 (F6H8S5) preservation of pancreases in a clinical setting compared with standard solutions for 1) cold ischemia time (CIT) <10 h and 2) an extended CIT >20 h. Procured clinical-grade pancreases were shipped in either F6H8S5 or in standard preservation solutions, that is, University of Wisconsin (UW) or Custodiol. F6H5S5 was preoxygenated for at least 15 min. Included clinical-grade pancreases were procured in UW or Custodiol. Upon arrival at the islet isolation laboratory, the duodenum was removed followed by rough trimming while F6H8S5 was oxygenated for 15–20 min. Trimmed pancreases were immersed into oxygenated F6H8S5 and stored at 4°C overnight followed by subsequent islet isolation. Pancreas preservation using F6H8S5 proved as effective as UW and Custadiol when used within CIT up to 10 h, in terms of both isolation outcome and islet functionality. Preservation in F6H8S5 of pancreases with extended CIT gave results similar to controls with CIT <10 h for both isolated islet functionality and isolation outcome. This study of clinically obtained pancreases indicates a clear benefit of using F6H8S5 on pancreases with extended CIT as it seems to allow extended cold ischemic time without affecting islet function and islet numbers.

Key words: Islet isolation; Pancreas preservation; Clinical islet transplantation; Perfluorohydrocarbons; Oxygenation; Hypoxia

Received November 3, 2015; final acceptance September 23, 2016. Online prepub date: May 27, 2016.
Address correspondence to Magnus Ståhle, IGP, Clinical Immunology, Rudbeck Laboratory, C11, Dag Hammarskjölds väg 20, SE-75185 Uppsala, Sweden. Tel: +46-70-4250507; Fax: +46-18-6110222; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 2269-2276, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691709
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved