Cell Transplantation 22(6) Abstracts

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Cell Transplantation, Vol. 22, pp. 921-944, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657477
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Direct Reprogramming of Adult Somatic Cells Into Other Lineages: Past Evidence and Future Perspectives

Monica Nizzardo, Chiara Simone, Marianna Falcone, Giulietta Riboldi, Giacomo P. Comi, Nereo Bresolin, and Stefania Corti

Department of Neurological Sciences, Dino Ferrari Centre, University of Milan, IRCCS Foundation Ca’ Granda, Ospedale Maggiore Policlinico, Milan, Italy

Direct reprogramming of an adult cell into another differentiated lineage—such as fibroblasts into neurons, cardiomyocytes, or blood cells—without passage through an undifferentiated pluripotent stage is a new area of research that has recently emerged alongside stem cell technology and induced pluripotent stem cell reprogramming; indeed, this avenue of investigation has begun to play a central role in basic biological research and regenerative medicine. Even though the field seems new, its origins go back to the 1980s when it was demonstrated that differentiated adult cells can be converted into another cell lineage through the overexpression of transcription factors, establishing mature cell plasticity. Here, we retrace transdifferentiation experiments from the discovery of master control genes to recent in vivo reprogramming of one somatic cell into another from the perspective of possible applications for the development of new therapeutic approaches for human diseases.

Key words: Transdifferentiation; Gene expression; Direct reprogramming

Received April 13, 2011; final acceptance April 18, 2012. Online prepub date: October 3, 2012.
Address correspondence to Stefania Corti, Department of Neurological Sciences, Dino Ferrari Centre, University of Milan, IRCCS Foundation Ca’ Granda, Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milan, Italy. Tel: +39-0255033817, Fax: +39-0250320430; Email: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 945-959, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657279
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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SSEA4-Positive Pig Induced Pluripotent Stem Cells Are Primed for Differentiation Into Neural Cells

Jeong-Yeh Yang,*† Jennifer L. Mumaw,*† Yubing Liu,*† Steve L. Stice,*† and Franklin D. West*†

*Regenerative Bioscience Center, University of Georgia, Rhodes Center for Animal and Dairy Science, Athens, GA, USA
†Department of Animal and Dairy Science, University of Georgia, Rhodes Center for Animal and Dairy Science, Athens, GA, USA

Neural cells derived from induced pluripotent stem cells (iPSCs) have the potential for autologous cell therapies in treating patients with severe neurological disorders or injury. However, further study of efficacy and safety are needed in large animal preclinical models that have similar neural anatomy and physiology to humans such as the pig. The pig model for pluripotent stem cell therapy has been made possible for the first time with the development of pig iPSCs (piPSCs) capable of in vitro and in vivo differentiation into tissues of all three germ layers. Still, the question remains if piPSCs are capable of undergoing robust neural differentiation using a system similar to those being used with human iPSCs. In this study, we generated a new line of piPSCs from fibroblast cells that expressed pluripotency markers and were capable of embryoid body differentiation into all three germ layers. piPSCs demonstrated robust neural differentiation forming βIII-TUB/MAP2+ neurons, GFAP+ astrocytes, and O4+ oligodendrocytes and demonstrated strong upregulation of neural cell genes representative of all three major neural lineages of the central nervous system. In the presence of motor neuron signaling factors, piPSC-derived neurons showed expression of transcription factors associated with motor neuron differentiation (HB9 and ISLET1). Our findings demonstrate that SSEA4 expression is required for piPSCs to differentiate into neurons, astrocytes, and oligodendrocytes and furthermore develop specific neuronal subtypes. This indicates that the pigs can fill the need for a powerful model to study autologous neural iPSC therapies in a system similar to humans.

Key words: Neural differentiation; Motor neuron; Induced pluripotent; Biomedical models

Received December 19, 2011; final acceptance May 23, 2012. Online prepub date: October 3, 2012.
Address correspondence to Franklin D. West, Regenerative Bioscience Center, Department of Animal and Dairy Science, University of Georgia, Rhodes Center for Animal and Dairy Science, 425 River Road, Athens, GA 30602-2771, USA. Tel: +1-706-542-0988; Fax: +1-706-542-7925; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 961-975, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657413
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Cholinergic Neuron-Like Cells Derived From Bone Marrow Stromal Cells Induced by Tricyclodecane-9-yl-xanthogenate Promote Functional Recovery and Neural Protection After Spinal Cord Injury

Chunhui Sun,*1 Jing Shao,*1 Le Su,*† Jing Zhao,*† Jianzhong Bi,‡ Shaonan Yang,‡ Shangli Zhang,*† Jiangang Gao,* and Junying Miao*†

*Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, Institute of Developmental Biology, School of Life Science, Shandong University, Jinan, China
†Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University, Qilu Hospital, Jinan, China
‡Department of Neural Medicine, The Second Hospital of Shandong University, Jinan, China

The rate of neuronal differentiation of bone marrow stromal cells (BMSCs) in vivo is very low; therefore, it is necessary to elevate the number of BMSC-derived neurons to cure neurodegenerative diseases. We previously reported that tricyclodecane-9-yl-xanthogenate (D609), an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), induced BMSCs to differentiate into neuron-like cells in vitro. However, the neuronal type is not clear, and it is still unknown whether these neuron-like cells possess physiological properties of functional neurons and whether they can contribute to the recovery of neuron dysfunction. To answer these questions, we investigated their characteristics by detecting neuronal function-related neurotransmitters and calcium image. The results showed that these cells exhibited functional cholinergic neurons in vitro. Transplantation of these cholinergic neuron-like cells promoted the recovery of spinal cord-injured mice, and they were more effective than BMSCs. The number of cholinergic neurons was increased after injection with BMSC-derived cholinergic neuron-like cells, indicating their high differentiation rate in vivo. Moreover, the proportion of cholinergic neurons in host cells and secretion of acetylcholine were increased, and preservation of neurofilament was also observed in the lesion of mice implanted with BMSC-derived neurons, suggesting the neuronal protection of BMSC-derived neurons. Our findings provide both a simple method to induce the differentiation of BMSCs into cholinergic neuron-like cells and a putative strategy for the therapy of spinal cord injuries.

Key words: Bone marrow stromal cells (BMSCs); Cholinergic neuron differentiation; Cell transplantation; Spinal cord injury (SCI); Tricyclodecane-9-yl-xanthogenate (D609)

Received June 14, 2011; final acceptance May 6, 2012. Online prepub date: October 1, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Prof. Junying Miao or Prof. Jiangang Gao, Institute of Developmental Biology, School of Life Science, Shandong University, Jinan 250100, China. Fax: +86 531 88565610; Tel: +86 531 88364929; E-mail address: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 977-991, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657251
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Delayed Intranasal Delivery of Hypoxic-Preconditioned Bone Marrow Mesenchymal Stem Cells Enhanced Cell Homing and Therapeutic Benefits After Ischemic Stroke in Mice

Ning Wei,*† Shan Ping Yu,* Xiaohuan Gu,* Tammi M. Taylor,* Denise Song,* Xin-feng Liu,† and Ling Wei*‡

*Department of Anesthesiology, Emory University School of Medicine, Atlanta, GA, USA
†Department of Neurology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, Jiangsu Province, People’s Republic of China
‡Department of Neurology, Emory University School of Medicine, Atlanta, GA, USA

Stem cell transplantation therapy has emerged as a potential treatment for ischemic stroke and other neurodegenerative diseases. Effective delivery of exogenous cells and homing of these cells to the lesion region, however, have been challenging issues that hinder the efficacy and efficiency of cell-based therapy. In the present investigation, we tested a delayed treatment of noninvasive and brain-targeted intranasal delivery of bone marrow mesenchymal stem cells (BMSCs) in a mouse focal cerebral ischemia model. The investigation tested the feasibility and effectiveness of intranasal delivery of BMSCs to the ischemic cortex. Hypoxia preconditioning (HP) of BMSCs was performed before transplantation in order to promote their survival, migration, and homing to the ischemic brain region after intranasal transplantation. Hoechst dye-labeled normoxic- or hypoxic-pretreated BMSCs (1 × 106 cells/animal) were delivered intranasally 24 h after stroke. Cells reached the ischemic cortex and deposited outside of vasculatures as early as 1.5 h after administration. HP-treated BMSCs (HP-BMSCs) showed a higher level of expression of proteins associated with migration, including CXC chemokine receptor type 4 (CXCR4), matrix metalloproteinase 2 (MMP-2), and MMP-9. HP-BMSCs exhibited enhanced migratory capacities in vitro and dramatically enhanced homing efficiency to the infarct cortex when compared with normoxic cultured BMSCs (N-BMSCs). Three days after transplantation and 4 days after stroke, both N-BMSCs and HP-BMSCs decreased cell death in the peri-infarct region; significant neuroprotection of reduced infarct volume was seen in mice that received HP-BMSCs. In adhesive removal test of sensorimotor functional assay performed 3 days after transplantation, HP-BMSC-treated mice performed significantly better than N-BMSC- and vehicle-treated animals. These data suggest that delayed intranasal administration of stem cells is feasible in the treatment of stroke and hypoxic preconditioning of transplanted cells, significantly enhances cell’s homing to the ischemic region, and optimizes the therapeutic efficacy.

Key words: Ischemic stroke; Bone marrow mesenchymal stem cells (BMSCs); Intranasal delivery; Hypoxic preconditioning; Cell homing

Received November 3, 2011; final acceptance May 18, 2012. Online prepub date: October 2, 2012.
Address correspondence to Ling Wei, M.D., 101 Woodruff Circle, Woodruff Memorial Research Building, Suite 617, Emory University School of Medicine, Atlanta, GA 30322, USA. Tel: +1-404-712-8661; Fax: +1-404-712-1351; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Xinfeng Liu, M.D., Ph.D., Department of Neurology, Jinling Hospital, Nanjing University School of Medicine, 305 East Zhongshan Road, Nanjing, Jiangsu Province, People’s Republic of China. Tel: +86 25 8537 2631; Fax: +86 25 8480 1861; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 993-1010, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657468
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Role of Matrix Metalloproteinases in Migration and Neurotrophic Properties of Nasal Olfactory Stem and Ensheathing Cells

Adlane Ould-Yahoui,*† Oualid Sbai,*† Kévin Baranger,*† Anne Bernard,*† Yatma Gueye,*† Eliane Charrat,*† Benoît Clément,‡§ Didier Gigmes,‡§ Vincent Dive,¶ Stéphane D. Girard,*† François Féron,*† Michel Khrestchatisky,*† and Santiago Rivera*†

*Aix-Marseille Univ, Neurobiologie des Interactions Cellulaires et Neurophysiopathologie (NICN), UMR 7259, 13344, Marseille, France

†CNRS, Neurobiologie des Interactions Cellulaires et Neurophysiopathologie (NICN), UMR 7259, 13344, Marseille, France
‡Aix-Marseille Univ, Institut de Chimie Radicalaire, Equipe Chimie Radicalaire, Organique et Polymères de Spécialité, UMR 7273, Marseille, France
§CNRS, Institut de Chimie Radicalaire, Equipe Chimie Radicalaire, Organique et Polymères de Spécialité, UMR 7273, Marseille, France
¶Département d’Ingénierie et d’Etudes des Protéines (DIEP), CEA/Saclay, Gif-sur-Yvette, France

Adult olfactory ectomesenchymal stem cells (OE-MSCs) and olfactory ensheathing cells (OECs), both from the nasal olfactory lamina propria, display robust regenerative properties when transplanted into the nervous system, but the mechanisms supporting such therapeutic effects remain unknown. Matrix metalloproteinases (MMPs) are an important family of proteinases contributing to cell motility and axonal outgrowth across the extracellular matrix (ECM) in physiological and pathological conditions. In this study, we have characterized for the first time in nasal human OE-MSCs the expression profile of some MMPs currently associated with cell migration and invasiveness. We demonstrate different patterns of expression for MMP-1, MMP-2, MMP-9, and MT1-MMP upon cell migration when compared with nonmigrating cells. Our results establish a correspondence between the localization of these proteinases in the migration front with the ability of cells to migrate. Using various modulators of MMP activity, we also show that at least MMP-2, MMP-9, and MT1-MMP contribute to OE-MSC migration in an in vitro 3D test. Furthermore, we demonstrate under the same conditions of culture used for in vivo transplantation that OE-MSCs and OECs secrete neurotrophic factors that promote neurite outgrowth of cortical and dorsal root ganglia (DRG) neurons, as well as axo-dendritic differentiation of cortical neurons. These effects were abolished by the depletion of MMP-2 and MMP-9 from the culture conditioned media. Altogether, our results provide the first evidence that MMPs may contribute to the therapeutic features of OE-MSCs and OECs through the control of their motility and/or their neurotrophic properties. Our data provide new insight into the mechanisms of neuroregeneration and will contribute to optimization of cell therapy strategies.

Key words: Adult olfactory stem cells; Olfactory ensheathing cells (OECs); Matrix metalloproteinases (MMPs); TIMPs; Cell therapy; Nervous system

Received January 24, 2012; final acceptance May 10, 2012. Online prepub date: October 3, 2012.
Address correspondence to Dr. Santiago Rivera, Neurobiologie des Interactions Cellulaires et Neurophysiopathologie (NICN), CNRS - Aix-Marseille Univ., Faculté de Médecine Secteur Nord, CS80011, 51 Bd Pierre Dramard, 13344 Marseille Cedex 15, France. Tel: +33 4 91 69 87 71; Fax: +33 4 91 25 89 70; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1011-1021, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657495
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Neurotrophin-3 Accelerates Wound Healing in Diabetic Mice by Promoting a Paracrine Response in Mesenchymal Stem Cells

Lei Shen,* Wen Zeng,† Yang-Xiao Wu,† Chun-Li Hou,† Wen Chen,† Ming-Can Yang,† Li Li,† Ya-Fang Zhang,*1 and Chu-Hong Zhu†1

*Department of Anatomy, Harbin Medical University, Harbin, China
†Department of Anatomy, Key Lab for Biomechanics of Chongqing, Third Military Medical University, Chongqing, China

Angiogenesis is a major obstacle for wound healing in patients with diabetic foot wounds. Mesenchymal stem cells (MSCs) have an important function in wound repair, and neurotrophin-3 (NT-3) can promote nerve regeneration and angiogenesis. We investigated the effect of NT-3 on accelerating wound healing in the diabetic foot by improving human bone marrow MSC (hMSC) activation. In vitro, NT-3 significantly promoted VEGF, NGF, and BDNF secretion in hMSCs. NT-3 improved activation of the hMSC conditioned medium, promoted human umbilical vein endothelial cell (HUVEC) proliferation and migration, and significantly improved the closure rate of HUVEC scratches. In addition, we produced nanofiber mesh biological tissue materials through the electrospinning technique using polylactic acid, mixed silk, and collagen. The hMSCs stimulated by NT-3 were implanted into the material. Compared with the control group, the NT-3-stimulated hMSCs in the biological tissue material significantly promoted angiogenesis in the feet of diabetic C57BL/6J mice and accelerated diabetic foot wound healing. These results suggest that NT-3 significantly promotes hMSC secretion of VEGF, NGF, and other vasoactive factors and that it accelerates wound healing by inducing angiogenesis through improved activation of vascular endothelial cells. The hMSCs stimulated by NT-3 can produce materials that accelerate wound healing in the diabetic foot and other ischemic ulcers.

Key words: Neurotrophin-3 (NT-3); Mesenchymal stem cells (MSCs); Diabetic foot; Paracrine; Vascular

Received November 15, 2011; final acceptance June 23, 2012. Online prepub date: October 3, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Chuhong Zhu, Department of Anatomy, Key Lab for Biomechanics of Chongqing, Third Military Medical University, Gao Tan Yan Street, Chongqing, 400038, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or


Cell Transplantation, Vol. 22, pp. 1023-1039, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653237
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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A Population of Selected Renal Cells Augments Renal Function and Extends Survival in the ZSF1 Model of Progressive Diabetic Nephropathy

Rusty Kelley,* Andrew Bruce,* Tom Spencer,* Eric Werdin,* Roger Ilagan,* Sumana Choudhury,* Elias Rivera,* Shay Wallace,* Kelly Guthrie,* Manuel Jayo,* Fengfeng Xu,† Anjali N. Rao,† Benjamin D. Humphreys,† Sharon Presnell,* and Tim Bertram*

*Tengion, Inc., Science and Technology, Winston-Salem, NC, USA
†Brigham and Women’s Hospital, Boston, MA, USA

New treatment paradigms that slow or reverse progression of chronic kidney disease (CKD) are needed to relieve significant patient and healthcare burdens. We have shown that a population of selected renal cells (SRCs) stabilized disease progression in a mass reduction model of CKD. Here, we further define the cellular composition of SRCs and apply this novel therapeutic approach to the ZSF1 rat, a model of severe progressive nephropathy secondary to diabetes, obesity, dyslipidemia, and hypertension. Injection of syngeneic SRCs into the ZSF1 renal cortex elicited a regenerative response that significantly improved survival and stabilized disease progression to renal structure and function beyond 1 year posttreatment. Functional improvements included normalization of multiple nephron structures and functions including glomerular filtration, tubular protein handling, electrolyte balance, and the ability to concentrate urine. Improvements to blood pressure, including reduced levels of circulating renin, were also observed. These functional improvements following SRC treatment were accompanied by significant reductions in glomerular sclerosis, tubular degeneration, and interstitial inflammation and fibrosis. Collectively, these data support the utility of a novel renal cell-based approach for slowing renal disease progression associated with diabetic nephropathy in the setting of metabolic syndrome, one of the most common causes of end-stage renal disease.

Key words: Diabetes; Regeneration; Renal; Survival; Nephropathy

Received January 26, 2012; final acceptance May 18, 2012. Online prepub date: August 10, 2012.
Address correspondence to Rusty Kelley, Ph.D., Director, Regenerative Medicine and Biology, Tengion, Inc., 3929 Westpoint Blvd., Suite G, Winston-Salem, NC 27103, USA. Tel: +1-(336)-722-5855; Fax: +1-(336)-722-2436; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1041-1051, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X655046
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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The Impact of Potential Islet Precursor Cells on Islet Autotransplantation Outcomes

M. A. Webb,* J. J. Chen,* S. C. Illouz,* C. A. Pollard,* B. Dennison,* K. P. West,† R. F. L. James,‡ and A. R. Dennison*

*Department of Hepatobiliary Surgery, University Hospitals of Leicester, NHS Trust, Leicester General Hospital, Leicester, UK
†Department of Pathology, University Hospitals of Leicester, NHS Trust, Leicester General Hospital, Leicester, UK
‡Department of Infection, Immunity and Inflammation, University of Leicester, Leicester, UK

Islet autotransplant patients represent excellent subjects to assess the posttransplant impact of islet precursors, as chronic pancreatitis (CP) causes an elevation of ductal cells, pancreatic precursors cells, and hormone-positive acinar cells. The relationship between these cell types and autograft outcomes should be more apparent than would be the case in the context of an allograft program with confounding immunological variables. To improve diabetic control following total pancreatectomy for CP, nonpurified islets were autotransplanted into the liver. Pancreas specimens were recovered from 23 patients and stained for antigens including: insulin, glucagon, cytokeratin 19, cytokeratin 7, and PDX-1. In line with previous reports, the prevalence of ductal cells, non-islet endocrine cells and non-islet PDX-1-expressing cells was significantly higher in CP glands compared with normal pancreata. When correlating follow-up data (i.e., fasting and stimulated C-peptide/glucose levels and HbA1c%) with pancreas immunoreactivity, high levels of ductal cells, non-islet PDX-1-positive cells, and non-islet glucagon-positive cells were associated with superior outcomes, detectable up to 2 years posttransplant. To conclude, the acinar parenchyma and ductal epithelium of the CP pancreas show an upregulation of both endocrine and pre-endocrine cell types, which appear to have a positive effect on islet graft outcomes in autotransplantation setting.

Key words: Islet autotransplantation; Chronic pancreatitis (CP); Ductal cells; Pancreatic and duodenal homeobox 1 (PDX-1); Glucagon

Received June 15, 2011; final acceptance March 4, 2012. Online prepub date: September 21, 2012.
Address correspondence to M’balu A. Webb, Department of Hepatobiliary Surgery, University Hospitals of Leicester NHS Trust, Leicester General Hospital, Gwendolen Road, Leicester, LE5 4PW, UK. Tel: +44 (0)116 258 8244; Fax: +44 (0)116 249 0064; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1053-1060, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657215
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Differentiation of Human Adipose Tissue-Derived Stem Cells Into Aggregates of Insulin-Producing Cells Through the Overexpression of Pancreatic and Duodenal Homeobox Gene-1

Jiyeon Lee,* Song Cheol Kim,*† Sung Jin Kim,‡ Heuiran Lee,‡§ Eun Jung Jung,* Seong Hee Jung,* and Duck Jong Han†

*Laboratory of Stem Cell Biology and Cell Therapy, Asan Institute for Life Sciences, Asan Medical Center, Seoul, Korea
†Department of Surgery, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea
‡Department of Microbiology, University of Ulsan College of Medicine, Seoul, Korea
§Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Seoul, Korea

The pancreatic and duodenal homeobox gene 1 (Pdx-1) plays a key role in normal pancreas development and is required for maintaining the normal function of islets. In this study, we examined whether human adipose tissue-derived stem cells (hASCs) could differentiate into insulin-producing cells by exogenously expressed Pdx-1. hASCs were infected with recombinant adenovirus encoding the mouse Pdx-1 gene and differentiated under high-glucose conditions. Insulin transcript levels and the expression of key transcription factors required for pancreatic development including FoxA2, Nkx2.2, and NeuroD were significantly increased by exogenous Pdx-1 overexpression. The expression of Nkx6.1 was found only in Pdx-1-induced hASCs. In addition to transcripts for transcription factors involved in pancreatic development, transcripts for the GLP-1 receptor, glucokinase, and glucose transporter, which are required for maintaining the function of pancreatic β-cells, were observed only in Pdx-1-induced hASCs. Pdx-1-induced hASCs exhibited insulin secretion in response to glucose challenge in vitro. When Pdx-1-induced hASCs were transplanted into streptozotocin (STZ)-induced diabetic mice, they reduced blood glucose levels, although they did not restore normoglycemia. These results demonstrate that the expression of exogenous Pdx-1 is sufficient to induce pancreatic differentiation in vitro but does not induce the fully functional, mature insulin-producing cells that are required for restoring normoglycemia in vivo.

Key words: Human adipose tissue-derived stem cells (hASCs); Pancreatic and duodenal homeobox gene (Pdx-1); Differentiation; Insulin-producing cells

Received July 12, 2011; final acceptance May 3, 2012. Online prepub date: October 1, 2012.
Address correspondence to Song Cheol Kim, Department of Surgery, University of Ulsan College of Medicine, Asan Medical Center, 86, Asanbyeongwon-gil, Songpa-gu, Seoul 138-736, Korea. Tel: +82-2-3010-3936; Fax: +82-2-474-9027; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1061-1073, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X656036
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Comparative Study of Human Dental Follicle Cell Sheets and Periodontal Ligament Cell Sheets for Periodontal Tissue Regeneration

Shujuan Guo,*†1 Weihua Guo,*1 Yi Ding,*† Jian Gong,*† Qing Zou,* Dan Xie,* Yali Chen,* Yafei Wu,*† and Weidong Tian*‡

*State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, Sichuan, China
†Department of Periodontology, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
‡Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China

Periodontal ligament cell (PDLC) sheets have been shown to contribute to periodontal tissue regeneration. Dental follicle cells (DFCs), acknowledged as the precursor cells of PDLCs, have demonstrated stemness, embryonic features, heterogeneity, and pluripotency. Therefore, we hypothesized that DFC sheets might be more effective and suitable for periodontal tissue regeneration than PDLC sheets. In this study, we compared the biological characteristics of DFC sheets and PDLC sheets in vitro. To investigate the potential for periodontal tissue regeneration in vivo, complexes composed of two types of cell sheets combined with dentin matrix were implanted subcutaneously into nude mice for 6 weeks. Our results showed that, when forming cell sheets, DFCs secreted richer extracellular matrix than PDLCs. And compared to DFCs, DFC sheets expressed high levels of calcification-related genes, including alkaline phosphatase (alp), bone sialoprotein (bsp), osteopontin (opn), runt-related transcription factor (runx2), as well as the periodontal ligament-specific genes collagen III (col III) and periostin, while the gene expression of bsp, osteocalcin (ocn), and opn were greatly increased in PDLC sheets, when compared to PDLCs. col I expression did not change significantly. However, cementum protein 23 (cp-23) expression increased several fold in PDLC sheets compared to PDLCs but decreased in DFC sheets compared to DFCs. DFC and PDLC sheets were both positive for Collagen I (Col I), cementum attachment protein (CAP), ALP, BSP, OCN, and OPN protein expression, and Col I, ALP, BSP, and OPN expression were increased after cell sheets were formed. Furthermore, the levels of laminin and fibronectin were higher in DFCs and DFC sheets than that of PDLCs and PDLC sheets, respectively. In vivo, DFC and PDLC sheets could both regenerate periodontal tissue-like structures, but DFC sheets demonstrated stronger periodontal regeneration potential than PDLC sheets. Therefore, DFC sheets derived from discarded dental follicle tissue after tooth extraction may be more advantageous for clinical periodontal tissue regeneration in the future.

Key words: Periodontal tissue regeneration; Cell sheets; Dental follicle cells (DFCs); Periodontal ligament cell (PDLCE)

Received August 24, 2011; final acceptance March 10, 2012. Online prepub date: September 21, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Weidong Tian, Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, No. 14, 3rd Section, Renmin Nan Road, Chengdu, Sichuan, China 610041. Tel: +86-28-85503499; Fax: +86-28-85503499; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Yafei Wu, Department of Periodontology, West China Hospital of Stomatology, Sichuan University, No. 14, 3rd Section, Renmin Nan Road, Chengdu, Sichuan, China 610041. Tel: +86-28-85503483; Fax: +86-28-85503499; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1075-1086, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657602
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
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Effects of Storage Solutions on the Viability of Human Umbilical Cord Mesenchymal Stem Cells for Transplantation

Yan Chen,* Baofeng Yu,† Guofang Xue,† Jie Zhao,† Ren-Ke Li,†‡ Zhuola Liu,* and Bo Niu†§

*Department of Respiratory Medicine, The Second Clinical Medical College, Shanxi Medical University, Taiyuan, China
†Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, China
‡Division of Cardiovascular Surgery, Department of Surgery, University of Toronto, Toronto, Ontario, Canada
§Biotechnology Research Laboratory, Capital Institute of Pediatrics, Beijing, China

Human umbilical cord-derived mesenchymal stem cell (UC-MSC) transplantation has shown promise for the treatment of various diseases. For clinical applications, UC-MSCs have been stored in 0.9% saline, 5% dextrose, dextrose and sodium chloride injection, Plasma-Lyte A, 1% human serum albumin (1% HSA), or 5% HSA before administration, but the effect of storage conditions on the viability and biological function of the cells remains unknown. Freshly harvested UC-MSCs were resuspended and incubated in these solutions for 2, 4, or 6 h at 4°C or room temperature (24°C). Cell viability, apoptotic/necrotic fraction, poststorage growth potential, immunophenotype, immunosuppressive capacity, and differentiation capacity were analyzed. When stored in parenteral solutions, UC-MSCs showed progressive deterioration in survival viability and adhesion ability. After 6-h storage, the best viability and attachment rate of UC-MSCs decreased to 83.0 ± 1.6% and 71.8 ± 3.2%, respectively. Our results suggested that UC-MSCs in these conditions lose their viability in a short time. However, it seems that the other biological functions of the surviving UC-MSCs were little affected. Since UC-MSCs suspended in these mediums lose their survival viability in a short time to levels significantly below the permissible limits (70%) by FDA, precautions need to be taken on using these solutions as suspension medium and further studies on the optimal methods for preservation are urgent.

Key words: Cell survival; Umbilical cord mesenchymal stem cells; Storage solutions; Transplantation; Effect

Received February 28, 2011; final acceptance May 7, 2012. Online prepub date: October 4, 2012.
Address correspondence to Dr. Zhuola Liu, Department of Respiratory Medicine, The Second Clinical Medical College, Shanxi Medical University, Taiyuan, P. R. China. Tel: +86-351-3365255; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Dr. Bo Niu, Biotechnology Research Laboratory, Capital Institute of Pediatrics, Beijing, P. R. China. Tel: +86-10-85695545; Fax: +86-10-62046502; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 22, pp. 1087-1099, 2013
0963-6897/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X657396
E-ISSN 1555-3892
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Evaluation of a Xeno-Free Protocol for Long-Term Cryopreservation of Cord Blood Cells

M. Mairhofer,*1 J. C. Schulz,†1 M. Parth,* U. Beer,* H. Zimmermann,†‡1 and A. Kolbus*1

*Department of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria
†Fraunhofer Institute for Biomedical Engineering (IBMT), St. Ingbert, Germany
‡University of Saarland, Chair of Molecular and Cellular Biotechnology/Nanotechnology, Saarbrücken, Germany

Cord blood is regarded as a powerful source for adult stem cells. Cord blood transplants have been used successfully to treat children and adults in autologous and allogeneic settings. Nevertheless, in many cases, the clinically relevant cell number (CD34+ cells and total leukocytes) is a limiting factor. To enable standardized cell banking and future in vitro expansion of adult stem/progenitor cells, elimination of serum, which inevitably differs from lot to lot and donor to donor, is highly desirable. Here, we demonstrate the feasibility of a xeno-free, chemically defined cryopreservation procedure for cord blood-derived cells over a period of 1 year. Cell recoveries with respect to retrieval of clinically relevant CD34+ cells, colony-forming units, and in vitro cultures of erythroid progenitor cells under standardized conditions were analyzed after 1 week or 1 year of cryopreservation and found to be very high and similar to the samples before freezing. The established xeno-free procedure is an important step toward using the full potential of adult stem cells from cord blood, enabling the elimination of serum-derived factors negatively influencing proliferation, differentiation, and survival of hematopoietic stem cells.

Key words: Cord blood; Optimization of cryopreservation; Hematopoietic stem/progenitor cells; Xeno-free cryomedium; Erythroid progenitor cells (EPCs)

Received May 25, 2011; final acceptance May 15, 2012. Online prepub date: October 3, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Mario Mairhofer, Department of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria. Tel: +43 1 40400-7692; Fax: +43 1 40400-7842; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it