Cell Transplantation 20(2) Abstracts

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Cell Transplantation, Vol. 20, pp. 145–152, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X522081
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Transplantation of Olfactory Ensheathing Cells as Adjunct Cell Therapy for Peripheral Nerve Injury

Christine Radtke,* Konstantin Wewetzer,†‡§ Kerstin Reimers,* and Peter M. Vogt*

*Department of Plastic, Hand- and Reconstructive Surgery, Hannover Medical School, Hannover, Germany
†Department of Pathology, University of Veterinary Medicine Hannover, Hannover, Germany
‡Department of Functional and Applied Anatomy, Center of Anatomy, Hannover Medical School, Hannover, Germany
§Center of Systems Neuroscience, Hannover, Germany

Traumatic events, such as work place trauma or motor vehicle accident violence, result in a significant number of severe peripheral nerve lesions, including nerve crush and nerve disruption defects. Transplantation of myelin-forming cells, such as Schwann cells (SCs) or olfactory ensheathing cells (OECs), may be beneficial to the regenerative process because the applied cells could mediate neurotrophic and neuroprotective effects by secretion of chemokines. Moreover, myelin-forming cells are capable of bridging the repair site by establishing an environment permissive to axonal regeneration. The cell types that are subject to intense investigation include SCs and OECs either derived from the olfactory bulb or the olfactory mucosa, stromal cells from bone marrow (mesenchymal stem cells, MSCs), and adipose tissue-derived cells. OECs reside in the peripheral and central nervous system and have been suggested to display unique regenerative properties. However, so far OECs were mainly used in experimental studies to foster central regeneration and it was not until recently that their regeneration-promoting activity for the peripheral nervous system was recognized. In the present review, we summarize recent experimental evidence regarding the regenerative effects of OECs applied to the peripheral nervous system that may be relevant to design novel autologous cell transplantation therapies.

Key words: Wallerian degeneration; Cell transplantation; Olfactory ensheathing cells (OECs); Regeneration; Remyelination; Cell expansion

Address correspondence to Christine Radtke, M.D., Department for Plastic, Hand- and Reconstructive Surgery, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany. Tel: ++49 511 532-8864; Fax: ++49 511 532-8890; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 153–166, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X522108
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

A Neuroregenerative Human Ensheathing Glia Cell Line With Conditional Rapid Growth

Vega García-Escudero,*† Ricardo Gargini,† María Teresa Gallego-Hernández,† Ana García-Gómez,*† María Jesús Martín-Bermejo,† Diana Simón,† Alicia Delicado,‡ María Teresa Moreno-Flores,† Jesús Ávila,† and Filip Lim*

*Departamento de Biología Molecular, Universidad Autónoma de Madrid, Cantoblanco, Madrid, Spain
†Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Cantoblanco, Madrid, Spain
‡Servicio de Genética Médica, Hospital La Paz, Madrid, Spain

Ensheathing glia have been demonstrated to have neuroregenerative properties but this cell type from human sources has not been extensively studied because tissue samples are not easily obtained, primary cultures are slow growing, and human cell lines are not available. We previously isolated immortalized ensheathing glia by gene transfer of BMI1 and telomerase catalytic subunit into primary cultures derived from olfactory bulbs of an elderly human cadaver donor. These cells escape the replicative senescence characteristic of primary human cells while conserving antigenic and neuroregenerative properties of ensheathing glia, but their low proliferative rate in culture complicates their utility as cell models and their application for preclinical cell therapy experiments. In this study we describe the use of a conditional SV40 T antigen (TAg) transgene to generate human ensheathing glia cell lines, which are easy to maintain due to their robust growth in culture. Although these fast growing clones exhibited polyploid karyotypes frequently observed in cells immortalized by TAg, they did not acquire a transformed phenotype, all of them maintaining neuroregenerative capacity and antigenic markers typical of ensheathing glia. These markers were also retained even after elimination of the TAg transgene using Cre/LoxP technology, although the cells died shortly after, confirming that their survival depended on the presence of the immortalizing genes. We have also demonstrated here the feasibility of using these human cell lines in animal models by genetically marking the cells with GFP and implanting them into the injured spinal cord of immunosuppressed rats. Our conditionally immortalized human ensheathing glia cell lines will thus serve as useful tools for advancing cell therapy approaches and understanding neuroregenerative mechanisms of this unique cell type.

Key words: Neuroregeneration; Spinal cord injury; Cell therapy; Olfactory ensheathing glia; Reversible immortalization; Cell expansion

Address correspondence to Filip Lim, Módulo 05 lab. 303, Francisco Tomás y Valiente, 7, Cantoblanco, 28049 Madrid, Spain. Tel: +34914977624; Fax: +34914973475; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 167–176, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X522090
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Optimal Time Point for Neuronal Generation of Transplanted Neural Progenitor Cells in Injured Spinal Cord Following Root Avulsion

Huanxing Su,* Yin Wu,† Qiuju Yuan,* Jiasong Guo,‡ Wenming Zhang,*§ and Wutian Wu*¶#**

*Department of Anatomy, The University of Hong Kong, Pokfulam, Hong Kong SAR, China
†Molecular Immunology Unit, University College London Institute of Child Health, London, UK
‡Department of Histology and Embryology, Southern Medical University, Guangzhou, China
§Department of Orthopaedics, The First Affiliated Hospital of Fujian Medical University, Fujian, China
¶State Key Laboratory of Brain and Cognitive Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China
#Research Center of Reproduction, Development and Growth, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China
**Joint Laboratory for Brain Function and Health (BFAH), Jinan University and The University of Hong Kong, Guangzhou, China

Root avulsion of the brachial plexus results in a progressive and pronounced loss of motoneurons. Cell replacement strategies have therapeutic potential in the treatment of motoneuron degenerative neurological disorders. Here, we transplanted spinal cord-derived neural progenitor cells (NPCs) into the cervical ventral horn of adult rats immediately, 2 weeks, or 6 weeks after root avulsion to determine an optimal time scale for the survival and differentiation of grafted cells. We showed that grafted NPCs survived robustly at all three time points and there was no statistical difference in survival rate. Interestingly, however, transplantation at 2 weeks postavulsion significantly increased the neuronal differentiation of transplanted NPCs compared to transplantation immediately or at 6 weeks postavulsion. Moreover, only NPCs transplanted at 2 weeks postavulsion were able to differentiate into choline acetyltransferase (ChAT)-positive neurons. Specific ELISAs and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) demonstrated that expression levels of BDNF and GDNF were significantly upregulated in the ventral cord at 2 weeks postavulsion compared to immediately or at 6 weeks postavulsion. Our study suggests that the cervical ventral horn at 2 weeks postavulsion both supports neuronal differentiation and induces region-specific neuronal generation possibly because of its higher expression of BDNF and GDNF.

Key words: Spinal root avulsion; Neural progenitor cells; Motoneurons; Transplantation; Neuronal differentiation

Address correspondence to Wutian Wu, Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Hong Kong SAR, China. Tel: (852) 28199187; Fax: (852) 28170857; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 177–191, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X527266
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Markers of Pluripotency and Differentiation in Human Neural Precursor Cells Derived From Embryonic Stem Cells and CNS Tissue

M. Sundberg,* P.-H. Andersson,† E. Åkesson,†‡ J. Odeberg,† L. Holmberg,† J. Inzunza,§ S. Falci,¶ J. Öhman,# R. Suuronen,*,**‡‡ H. Skottman,* K. Lehtimäki,# O. Hovatta,‡‡ S. Narkilahti,* and E. Sundström†‡

*Regea-Institute for Regenerative Medicine, University of Tampere and Tampere University Hospital, Tampere, Finland
†Division of Neurodegeneration, Department of Neurobiology, Care Sciences and Society, Karolinska Institutet, Stockholm, Sweden
‡Stockholms Sjukhem Foundation, Stockholm, Sweden
§Department of Biosciences and Nutrition, Karolinska Institute, Stockholm, Sweden
¶Department of Neurosurgery, Craig Hospital, Denver, CO, USA
#The Pirkanmaa Hospital District, Tampere University Hospital, Tampere, Finland
**Department of Eye, Ear, and Oral Diseases, Tampere University Hospital, Tampere, Finland
††Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland
‡‡Division of Obstetrics and Gynecology, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden

Cell transplantation therapies for central nervous system (CNS) deficits such as spinal cord injury (SCI) have been shown to be effective in several animal models. One cell type that has been transplanted is neural precursor cells (NPCs), for which there are several possible sources. We have studied NPCs derived from human embryonic stem cells (hESCs) and human fetal CNS tissue (hfNPCs), cultured as neurospheres, and the expression of pluripotency and neural genes during neural induction and in vitro differentiation. mRNA for the pluripotency markers Nanog, Oct-4, Gdf3, and DNMT3b were downregulated during neural differentiation of hESCs. mRNA for these markers was found in nonpluripotent hfNPC at higher levels compared to hESC-NPCs. However, Oct-4 protein was found in hESC-NPCs after 8 weeks of culture, but not in hfNPCs. Similarly, SSEA-4 and CD326 were only found in hESC-NPCs. NPCs from both sources differentiated as expected to cells with typical features of neurons and astrocytes. The expressions of neuronal markers in hESC-NPCs were affected by the composition of cell culture medium, while this did not affect hfNPCs. Transplantation of hESC-NPC or hfNPC neurospheres into immunodeficient mouse testis or subcutaneous tissue did not result in tumor formation. In contrast, typical teratomas appeared in all animals after transplantation of hESC-NPCs to injured or noninjured spinal cords of immunodeficient rats. Our data show that transplantation to the subcutaneous tissue or the testes of immunodeficient mice is not a reliable method for evaluation of the tumor risk of remaining pluripotent cells in grafts.

Key words: Neural precursor cells; Embryonic stem cells; Pluripotency; Spinal cord injury; Transplantation; Tumor

Address correspondence to Erik Sundström, Division of Neurodegeneration, Geriatric Clinic Res Lab, Novum 5th Floor, S14186 Stockholm, Sweden. Tel: +46 858583887; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 193–203, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X514305
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Late Passage Human Fibroblasts Induced to Pluripotency Are Capable of Directed Neuronal Differentiation

Jun Liu,* Huseyin Sumer,* Jessie Leung,† Kyle Upton,* Mirella Dottori,†‡ Alice Pébay,†‡ and Paul J. Verma*

*Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia
†Centre for Neuroscience, The University of Melbourne, Parkville, Victoria, Australia
‡Department of Pharmacology, The University of Melbourne, Parkville, Victoria, Australia

It is possible to generate induced pluripotent stem (iPS) cells from mouse and human somatic cells by ectopic expression of defined sets of transcription factors. However, the recommendation that somatic cells should be utilized at early passages for induced reprogramming limits their therapeutic application. Here we report successful reprogramming of human fibroblasts after more than 20 passages in vitro, to a pluripotent state with four transcription factors: Oct4, Sox2, Klf4, and c-Myc. The late passage-derived human iPS cells resemble human embryonic stem cells in morphology, cell surface antigens, pluripotent gene expression profiles, and epigenetic states. Moreover, these iPS cells differentiate into cell types representative of the three germ layers in teratomas in vivo, and directed neuronal differentiation in vitro.

Key words: Pluripotent reprogramming; Late passage; Human fibroblasts; Stem cell; Neuronal differentiation

Address correspondence to Paul Verma, 27-31 Wright Street, Clayton VIC 3168, Australia. Tel: 61 3 9594 7000; Fax: 61 3 9594 7416; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 205–216, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X520065
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
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Locally Administered Adipose-Derived Stem Cells Accelerate Wound Healing Through Differentiation and Vasculogenesis

Chunlei Nie,* Daping Yang,† Jin Xu,‡ Zhenxing Si,† Xiaoming Jin,§ and Jiewu Zhang*

*Department of Head and Neck Surgery, The Third Affiliated Hospital of Harbin Medical University, Harbin, China
†Department of Plastic Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, China
‡Department of Cell Biology, Harbin Medical University, Harbin, China
§Department of Pathology, Harbin Medical University, Harbin, China

Despite advances in wound closure techniques and devices, there is still a critical need for new methods of enhancing the healing process to achieve optimal outcomes. Recently, stem cell therapy has emerged as a new approach to accelerate wound healing. Adipose-derived stem cells (ASCs) hold great promise for wound healing, because they are multipotential stem cells capable of differentiation into various cell lineages and secretion of angiogenic growth factors. The aim of this study was to evaluate the benefit of ASCs on wound healing and then investigate the probable mechanisms. ASCs characterized by flow cytometry were successfully isolated and cultured. An excisional wound healing model in rat was used to determine the effects of locally administered ASCs. The gross and histological results showed that ASCs significantly accelerated wound closure in normal and diabetic rat, including increased epithelialization and granulation tissue deposition. Furthermore, we applied GFP-labeled ASCs on wounds to determine whether ASCs could differentiate along multiple lineages of tissue regeneration in the specific microenvironment. Immunofluorescent analysis indicated that GFP-expressing ASCs were costained with pan-cytokeratin and CD31, respectively, indicating spontaneous site-specific differentiation into epithelial and endothelial lineages. These data suggest that ASCs not only contribute to cutaneous regeneration, but also participate in new vessels formation. Moreover, ASCs were found to secret angiogenic cytokines in vitro and in vivo, including VEGF, HGF, and FGF2, which increase neovascularization and enhance wound healing in injured tissues. In conclusion, our results demonstrate that ASC therapy could accelerate wound healing through differentiation and vasculogenesis and might represent a novel therapeutic approach in cutaneous wounds.

Key words: Wound healing; Adipose-derived stem cells (ASCs); Differentiation; Secretion; Vascularization

Address correspondence to Jiewu Zhang, Department of Head and Neck Surgery, The Third Affiliated Hospital of Harbin Medical University, 150 Ha Ping Road, Harbin 150040, China. Tel: 86-451-86298333; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Daping Yang, Department of Plastic Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 217–231, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X522117
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Long-Term Contribution of Human Bone Marrow Mesenchymal Stromal Cells to Skeletal Muscle Regeneration in Mice

Anabel S. de la Garza-Rodea, Ietje van der Velde, Hester Boersma, Manuel A. F. V. Gonçalves, Dirk W. van Bekkum, Antoine A. F. de Vries, and Shoshan Knaän-Shanzer

Virus and Stem Cell Biology Laboratory, Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, the Netherlands

Mesenchymal stromal cells (MSCs) are attractive for cellular therapy of muscular dystrophies as they are easy to procure, can be greatly expanded ex vivo, and contribute to skeletal muscle repair in vivo. However, detailed information about the contribution of bone marrow (BM)-derived human MSCs (BM-hMSCs) to skeletal muscle regeneration in vivo is very limited. Here, we present the results of a comprehensive study of the fate of LacZ-tagged BM-hMSCs following implantation in cardiotoxin (CTX)-injured tibialis anterior muscles (TAMs) of immunodeficient mice. β-Galactosidase-positive (β-gal+) human–mouse hybrid myofibers (HMs) were counted in serial cross sections over the full length of the treated TAMs of groups of mice at monthly intervals. The number of human cells was estimated using chemiluminescence assays. While the number of human cells declined gradually to about 10% of the injected cells at 60 days after transplantation, the number of HMs increased from day 10 onwards, reaching 104 ± 39.1 per TAM at 4 months postinjection. β-gal+ cells and HMs were distributed over the entire muscle, indicating migration of the former from the central injection site to the ends of the TAMs. The identification of HMs that stained positive for human spectrin suggests myogenic reprogramming of hMSC nuclei. In summary, our findings reveal that BMhMSCs continue to participate in the regeneration/remodeling of CTX-injured TAMs, resulting in ±5% HMs at 4 months after damage induction. Moreover, donor-derived cells were shown to express genetic information, both endogenous and transgenic, in recipient myofibers.

Key words: Bone marrow; Cell therapy; Duchenne muscular dystrophy; Human mesenchymal stromal cell; NOD/SCID mouse; Skeletal muscle regeneration

Address correspondence to Shoshan Knaän-Shanzer, Ph.D., Virus and Stem Cell Biology Laboratory, Department of Molecular Cell Biology, Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, the Netherlands. Tel: +31(71) 526 9246; Fax: +31 (71) 526 8270; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 233–243, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X522234
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Beneficial Effect of Autologous Transplantation of Endothelial Progenitor Cells on Steroid-Induced Femoral Head Osteonecrosis in Rabbits

Yuan Sun, Yong Feng, Changqing Zhang, Xiangguo Cheng, Shengbao Chen, Zisheng Ai, and Bingfang Zeng

Department of Orthopaedics, Shanghai Sixth People’s Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, China

Femoral head osteonecrosis (ON) is a serious complication of steroid administration. We examined whether implantation of autologous bone marrow-derived endothelial progenitor cells (EPCs) can augment neovascularization and bone regeneration in steroid-induced osteonecrosis of the femoral head. Forty 12-week-old male New-Zealand white rabbits were divided into group I (left untreated, n = 12), group II (core decompression, n = 12), and group III (core decompression + autologous EPCs implantation, n = 16) after receiving an established inductive protocol for inducing steroid-associated ON. Four weeks later, these rabbits were euthanized, bilateral femora were dissected for Micro-CT-based microangiography to assess vascularization, and then the osteonecrotic changes and repair processes were examined histopathologically. Quantitative analysis showed that new vessel formation in group III was significantly greater compared with other groups at 4 weeks after treatment. The histologic and histomorphometric analyses revealed that the new bone volume was significantly higher in group III than in groups I and II 4 weeks after treatment. A combination of EPCs and core decompression enhances the neovascularization and bone regeneration in rabbit steroidinduced femoral head ON. Local implantation of EPCs may provide a novel and effective therapeutic option for early corticosteroid-induced ON.

Key words: Endothelial progenitor cells (EPCs); Steroid; Osteonecrosis; Vascularization; Bone regeneration

Address correspondence to Changqing Zhang, Department of Orthopaedics, Shanghai Sixth People?s Hospital, 600 Yishan Road, Shanghai Jiao Tong University, School of Medicine, 200233 Shanghai, China. Tel: +8621-64369181; Fax: +8621-64369181; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 245–257, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X520056
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Therapeutic Effects of Umbilical Cord Blood-Derived Mesenchymal Stem Cell Transplantation in Experimental Lupus Nephritis

Jei-Wen Chang,*,** Shun-Pei Hung,† Hao-Hsiang Wu,† Wen-Mien Wu,‡ An-Hang Yang,§** Hsin-Lin Tsai,¶** Ling-Yu Yang,*,** and Oscar K. Lee*

*Division of Immunology and Nephrology, Department of Pediatrics, Taipei Veterans General Hospital, Taipei, Taiwan
†Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan
‡Department of Nutrition and Food Sciences, Fu-Jen University, Taipei, Taiwan
§Division of Ultrastructural and Molecular Pathology, Department of Pathology, Taipei Veterans General Hospital, Taipei, Taiwan
¶Division of Pediatric Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
#Department of Orthopedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan
**Institute of Clinical Medicine, National Yang-Ming University, School of Medicine, Taipei, Taiwan

Mesenchymal stem cells (MSCs) have been shown to possess immunomodulatory properties. Systemic lupus erythematosus is an autoimmune disease that results in nephritis and subsequent destruction of renal microstructure. We investigated whether transplantation of human umbilical cord blood-derived MSCs (uMSCs) is useful in alleviating lupus nephritis in a murine model. It was found that uMSCs transplantation significantly delayed the development of proteinuria, decreased anti-dsDNA, alleviated renal injury, and prolonged the life span. There was a trend of decreasing T-helper (Th) 1 cytokines (IFN-γ, IL-2) and proinflammatory cytokines (TNF-α, IL-6, IL-12) and increasing Th2 cytokines (IL-4, IL-10). The in vitro coculture experiments showed that uMSCs only inhibited lymphocytes and splenocytes proliferation but not mesangial cells. Long-term engraftment of uMSCs in the kidney was not observed either. Together, these findings indicated that uMSCs were effective in decreasing renal inflammation and alleviating experimental lupus nephritis by inhibiting lymphocytes, inducing polarization of Th2 cytokines, and inhibition of proinflammatory cytokines production rather than direct engraftment and differentiating into renal tissue. Therapeutic effects demonstrated in this preclinical study support further exploration of the possibility to use uMSCs from mismatched donors in lupus nephritis treatment.

Key words: Immunomodulation; Lupus nephritis; Umbilical cord blood; Mesenchymal stem cells

Address correspondence to Oscar K. Lee, Department of Orthopedics and Traumatology, Taipei Veterans General Hospital, Taiwan and Institute of Clinical Medicine, National Yang-Ming University, School of Medicine, Taiwan, No. 201, Shi-Pai Rd., Sec. 2, Peitou, Taipei 11217, Taiwan. Tel: +886-2-2875-7557; Fax: +886-2-2875-7657; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 259–269, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X516592
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
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MAPC Transplantation Confers a More Durable Benefit Than AC133+ Cell Transplantation in Severe Hind Limb Ischemia

Xabier L. Aranguren,*‡1 Beatriz Pelacho,*1 Ivan Peñuelas,† Gloria Abizanda,* Maialen Uriz,* Margarita Ecay,† María Collantaes,† Miriam Araña,* Manu Beerens,‡ Giulia Coppiello,‡ Inés Prieto,* Maitane Perez-Ilzarbe,* Enrique J. Andreu,* Aernout Luttun,‡ and Felipe Prósper*

*Hematology Service and Cell Therapy, Foundation for Applied Medical Research, Division of Cancer, University of Navarra, Pamplona, Spain
†Department of Nuclear Medicine, Clínica Universitaria, University of Navarra, Pamplona, Spain
‡Center for Molecular and Vascular Biology, Catholic University of Leuven, Leuven, Belgium

There is a need for comparative studies to determine which cell types are better candidates to remedy ischemia. Here, we compared human AC133+ cells and multipotent adult progenitor cells (hMAPC) in a mouse model reminiscent of critical limb ischemia. hMAPC or hAC133+ cell transplantation induced a significant improvement in tissue perfusion (measured by microPET) 15 days posttransplantation compared to controls. This improvement persisted for 30 days in hMAPC-treated but not in hAC133+-injected animals. While transplantation of hAC133+ cells promoted capillary growth, hMAPC transplantation also induced collateral expansion, decreased muscle necrosis/fibrosis, and improved muscle regeneration. Incorporation of differentiated hAC133+ or hMAPC progeny into new vessels was limited; however, a paracrine angio/arteriogenic effect was demonstrated in animals treated with hMAPC. Accordingly, hMAPC-conditioned, but not hAC133+-conditioned, media stimulated vascular cell proliferation and prevented myoblast, endothelial, and smooth muscle cell apoptosis in vitro. Our study suggests that although hAC133+ cell and hMAPC transplantation both contribute to vascular regeneration in ischemic limbs, hMAPC exert a more robust effect through trophic mechanisms, which translated into collateral and muscle fiber regeneration. This, in turn, conferred tissue protection and regeneration with longer term functional improvement.

Key words: Angiogenesis; Stem cells; Critical limb ischemia; Multipotent adult progenitor cells (MAPC); AC133+ cells

1These authors provided equal contribution to this work.
Address correspondence to Felipe Pro´sper, Hematology and Cell Therapy Area, Clínica Universitaria, University of Navarra, Av Pio XII 36, Pamplona 31009, Spain. Tel: +34-948-255400; Fax: +34-948-296500; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Aernout Luttun, Center for Molecular and Vascular Biology, Katholieke Universiteit Leuven, Campus Gasthuisberg, Herestraat 49, B-3000 Leuven, Belgium. Tel: +32-16-34-57-72; Fax: +32-16-34-59-90; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 271–285, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X519292
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Efficacy of Periodontal Stem Cell Transplantation in the Treatment of Advanced Periodontitis

Joo-Young Park,*†1 Soung Hoo Jeon,*†1 and Pill-Hoon Choung*†

*Department of Oral and Maxillofacial Surgery and Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Republic of Korea
†Tooth Bioengineering National Research Laboratory, BK21, School of Dentistry, Seoul National University, Seoul, Republic of Korea

Periodontitis is the most common cause for tooth loss in adults and advanced types affect 10–15% of adults worldwide. The attempts to save tooth and regenerate the periodontal apparatus including cementum, periodontal ligament, and alveolar bone reach to the dental tissue-derived stem cell therapy. Although there have been several periodontitis models suggested, the apical involvement of tooth root is especially challenging to be regenerated and dental stem cell therapy for the state has never been investigated. Three kinds of dental tissue-derived adult stem cells (aDSCs) were obtained from the extracted immature molars of beagle dogs (n = 8), and ex vivo expanded periodontal ligament stem cells (PDLSCs), dental pulp stem cells (DPSCs), and periapical follicular stem cells (PAFSCs) were transplanted into the apical involvement defect. As for the lack of cementum-specific markers, anti-human cementum protein 1 (rhCEMP1) antibody was fabricated and the aDSCs and the regenerated tissues were immunostained with anti-CEMP1 antibody. Autologous PDLSCs showed the best regenerating capacity of periodontal ligament, alveolar bone, and cementum as well as peripheral nerve and blood vessel, which were evaluated by conventional and immune histology, 3D micro-CT, and clinical index. The rhCEMP1 was expressed strongest in PDLSCs and in the regenerated periodontal ligament space. We suggest here the PDLSCs as the most favorable candidate for the clinical application among the three dental stem cells and can be used for treatment of advanced periodontitis where tooth removal was indicated in the clinical cases.

Key words: Dental mesenchymal stem cell; Adult stem cell therapy; Advanced periodontitis; Tooth; Cementum

1These authors provided equal contribution to this work.
Address correspondence to Pill-Hoon Choung, D.D.S., Ph.D., Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, 28 Yeongeon-dong, Jongno-gu, Seoul, Republic of Korea, 110-749. Tel: +82-2-740-8717; Fax: +82-2-740-8717; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 167–176, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X522090
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

In Vitro Differentiated Adult Human Liver Progenitor Cells Display Mature Hepatic Metabolic Functions: A Potential Tool for In Vitro Pharmacotoxicological Testing

Dung Ngoc Khuu,* Isabelle Scheers,* Sabrina Ehnert,† Nawal Jazouli,* Omar Nyabi,* Pedro Buc-Calderon,‡ Ann Meulemans,§ Andreas Nussler,† Etienne Sokal,* and Mustapha Najimi*

*Institut de Recherche Clinique et Expérimentale (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Université Catholique de Louvain, Brussels, Belgium
†Department of Traumatology, Technical University Munich, Munich, Germany
‡Louvain Drug Research Institute, Toxicology and Cancer Biology Research Group, PMNT Unit, Université Catholique de Louvain, Brussels, Belgium
§Laboratoire de Pédiatrie, Unité Métabolique, Université Libre de Bruxelles, Brussels, Belgium

The potential use of stem/progenitor cells as alternative cell sources to mature hepatocytes remains basically dependent on their ability to exhibit some, if not all, the metabolic liver functions. In the current study, four major liver functions were investigated in adult derived human liver stem/progenitor cell (ADHLSCs) populations submitted to in vitro hepatogenic differentiation: gluconeogenesis, ammonia detoxification, and activity of phase I and phase II drug-metabolizing enzymes. These acquired hepatic activities were compared to those of primary adult human hepatocytes, the standard reference. Amino acid content was also investigated after hepatogenic differentiation. Differentiated ADHLSCs display higher de novo synthesis of glucose correlated to an increased activity of glucose-6 phosphatase and mRNA expression of key related enzymes. Differentiated ADHLSCs are also able to metabolize ammonium chloride and to produce urea. This was correlated to an increase in the mRNA expression of relevant key enzymes such arginase. With respect to drug metabolism, differentiated ADHLSCs express mRNAs of all the major cytochromes investigated, among which the CYP3A4 isoform (the most important drug-metabolizing enzyme). Such increased expression is correlated to an enhanced phase I activity as independently demonstrated using fluorescence-based assays. Phase II enzyme activity and amino acid levels also show a significant enhancement in differentiated ADHLSCs. The current study, according to data independently obtained in different labs, demonstrates that in vitro differentiated ADHLSCs are able to display advanced liver metabolic functions supporting the possibility to develop them as potential alternatives to primary hepatocytes for in vitro settings.

Key words: Mesenchymal cells; Hepatic stem cells; Hepatocyte differentiation; Metabolic activity; Tissue-specific stem cells

Address correspondence to Mustapha Najimi, Ph.D., Laboratory of Pediatric Hepatology and Cell Therapy, Avenue Hippocrate 10/1301,1200, Brussels, Belgium. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 303–311, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X520074
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Regional Transient Portal Ischemia and Irradiation as Preparative Regimen for Hepatocyte Transplantation

S. Koenig,* Q. Yuan,†‡ P. Krause,* H. Christiansen,§ M. Rave-Fraenk,§ S. Kafert-Kasting,¶ H. Kriegbaum,¶ A. Schneider,† M. Ott,†‡1 and J. Meyburg#1

*Department of General and Visceral Surgery, University Medical Centre Goettingen, Goettingen, Germany
†Department of Gastroenterology, Hepatology and Endocrinology, Centre of Internal Medicine, Hanover Medical School, Hanover, Germany
‡Twincore Centre for Experimental and Clinical Research, Hannover, Germany
§Department of Radiotherapy, University Medical Centre Goettingen, Goettingen, Germany
¶Cytonet GmbH & Co. KG, Weinheim, Germany
#Department of General Paediatrics, University Children’s Hospital, Heidelberg, Germany

Hepatocyte transplantation is regarded as a promising option to correct hereditary metabolic liver disease. This study describes a novel method involving regional transient portal ischemia (RTPI) in combination with hepatic irradiation (IR) as a preparative regimen for hepatocyte transplantation. The right lobules of rat livers (45% of liver mass) were subjected to RTPI of 30–120 min. Liver specimens and serum samples were analyzed for transaminase levels, DNA damage, apoptosis, and proliferation. Repopulation experiments involved livers of dipeptidylpeptidase IV (DPPIV)-deficient rats preconditioned with RTPI (60–90 min) either with or without prior partial hepatic IR (25 Gy). After reperfusion intervals of 1 and 24 h, 12 million wild-type (DPPIV positive) hepatocytes were transplanted into recipient livers via the spleen. RTPI of 60–90 min caused limited hepatic injury through necrosis and induced a distinct regenerative response in the host liver. Twelve weeks following transplantation, small clusters of donor hepatocytes were detected within the portal areas. Quantitative analysis revealed limited engraftment of 0.79% to 2.95%, whereas control animals (sham OP) exhibited 4.16% (determined as relative activity of DPPIV when compared to wild-type liver). Repopulation was significantly enhanced (21.43%) when IR was performed prior to RTPI, optimum preconditioning settings being 90 min of ischemia and 1 h of reperfusion before transplantation. We demonstrate that RTPI alone is disadvantageous to donor cell engraftment, whereas the combination of IR with RTPI comprises an effective preparative regimen for liver repopulation. The method described clearly has potential for clinical application.

Key words: Irradiation; Ischemia reperfusion injury; Hepatocyte transplantation; Preconditioning; Liver repopulation

1Shared last authorship.
Address correspondence to PD Dr. med. Sarah Koenig, Department of General and Visceral Surgery, University Medical Centre Goettingen, Robert-Koch-Strasse 40, 37099 Goettingen, Germany. Tel: +49-551-398977; Fax: +49-551-396107; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 313–322, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X514611
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Body Mass Index Reflects Islet Isolation Outcome in Islet Autotransplantation for Patients With Chronic Pancreatitis

Morihito Takita,* Bashoo Naziruddin,† Shinichi Matsumoto,* Hirofumi Noguchi,* Masayuki Shimoda,‡ Daisuke Chujo,§ Takeshi Itoh,* Koji Sugimoto,* Yoshiko Tamura,† Greg S. Olsen,† Nicholas Onaca,† Jeffrey Lamont,‡ Luis F. Lara,‡ and Marlon F. Levy†

*Baylor Research Institute Fort Worth Campus, Fort-Worth, TX, USA
†Baylor Regional Transplant Institute, Dallas, TX, USA
‡Baylor University Medical Center, Dallas, TX, USA
§Baylor Institute for Immunology Research, Dallas, TX, USA

Total pancreatectomy with autologous islet cell transplantation (TP with AIT) is an effective treatment for chronic pancreatitis patients with severe abdominal pain. Body mass index (BMI) of the pancreatic donor is proven to be a useful predictor for islet isolation and transplantation outcomes in allogenic islet transplantation. However, the association between BMI and islet isolation outcome and/or metabolism after AIT was previously unclear. Twelve patients who received TP with AIT at our hospital were included in this study. All pancreata were preserved with both pancreatic ductal injection and oxygen-charged static two-layer method using ET-Kyoto solution. The cohort was divided into two groups: low BMI group (BMI < 23 kg/m2, n = 5) and high BMI group (BMI ≥ 23, n = 7). The high BMI group had a significantly higher islet yield per gram than the low BMI group both in pancreas postdigestion and in final product (postdigestion: 7330 ± 539 vs. 3509 ± 563 IE/g; p < 0.001; final product: 6555 ± 585 vs. 3476 ± 546 IE/g; p = 0.004). For islet yield in final product per patient body weight, the high BMI group also had significantly higher islet yield than the low BMI group (7997 ± 779 vs. 4175 ± 750 IE/kg, p = 0.007). Insulin independence rate in the high BMI group (71%) was also higher than that low BMI group (40%), but it did not reach statistical significance. Pancreata from patients with higher BMI could obtain higher islet yield in the setting of autologous islet cell transplantation for chronic pancreatitis.

Key words: Body mass index (BMI); Progression of inflammation; Islet isolation

Address correspondence to Shinichi Matsumoto, Islet Cell Laboratory, Baylor Research Institute Fort Worth Campus, 1400 8th Avenue, Fort Worth, TX 76104, USA. Tel: 817-922-2570; Fax: 817-922-4645; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 323–332, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X514189
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Use of Perfluorodecalin for Pancreatic Islet Culture Prior to Transplantation: A Liquid–Liquid Interface Culture System—Preliminary Report

M. T. Juszczak,*† A. Elsadig,* A. Kumar,*† M. Muzyamba,† K. Pawelec,† S. H. Powis,† and M. Press*

*Department of Endocrinology and Diabetes, Royal Free Hospital, Hempsted, London, UK
†Centre for Nephrology, Hampstead Campus, University College London, Hampstead, London, UK

Although the issue remains controversial, short-term culture is probably beneficial for islet graft quality. However, significant islet loss is invariably observed. This is related to reduced survival of large islets, which is compromised by hypoxia under standard culture conditions. We aimed to develop a method of culture, which would avoid exposure to relative hypoxia and hence maintain the quality of islets. Isolated rat islets cultured for 48 h in a liquid–liquid interface culture system (LICS) with a perfluorocarbon were compared to islets cultured under standard (C1) and suboptimal conditions (C2). Islets were tested for viability and response to a glucose challenge, and a marginal mass was transplanted into syngeneic diabetic recipients. The viability of islets after 24-h culture in LICS was higher than in C1 and C2 groups (89.0% vs. 77.5% and 64.6%, respectively) and decreased with time to reach 79.0%, 62.9%, and 53.4% after 72-h culture. The stimulation index in LICS-cultured islets was also significantly higher than in C1 and C2 groups (12.3 ± 0.4 vs. 5.8 ± 0.5 and 4.1 ± 0.2, respectively). Following transplantation of LICS-cultured islets 50% of recipients were rendered normoglycemic compared with 14.3% and 31.3% for C2 and fresh islets, respectively. Our liquid–liquid interface culture system using perfluorodecalin provides optimized culture conditions, which preserve both islet viability and their ability to engraft successfully after intraportal transplantation and could be used for islet transportation.

Key words: Rat; Pancreatic islets; Syngeneic transplantation; Perfluorocarbons; Cell culture; Hypoxia

Address correspondence to Dr. Maciej Juszczak, Ph.D., Centre for Nephrology, Hampstead Campus, University College London, Rowland Hill Street, Hampstead, London NW3 2PF, UK. Tel/Fax: +44 20 7830 2475; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 333–350, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X552844
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Alternatively Expressed Genes Identified in the CD4+ T Cells of Allograft Rejection Mice

Jia Xu,*1 Dan Wang,*1 Chao Zhang,* Jing Song,* Ting Liang,* Weirong Jin,† Yeong C. Kim,‡ San Ming Wang,‡ and Guihua Hou*

*Key Laboratory for Experimental Teratology of the Ministry of Education and Institute of Experimental Nuclear Medicine, School of Medicine, Shandong University, Jinan, Shandong, P.R. China
†Shanghai Huaguan BioChip Co., Ltd, Shanghai, P.R. China
‡Department of Genetics, Cell Biology & Anatomy, College of Medicine, University of Nebraska Medical Center, Omaha, NE, USA

Allograft rejection is a leading cause for the failure of allotransplantation. CD4+ T cells play critical roles in this process. The identification of genes that alternatively expressed in CD4+ T cells during allograft rejection will provide critical information for studying the mechanism of allograft rejection, finding specific gene markers for monitoring, predicting allograft rejection, and opening new ways to regulate and prevent allograft rejection. Here, we established allograft and isograft transplantation models by adoptively transferring wild-type BALB/c mouse CD4+ T cells into severe combined immunodeficient (SCID) mice with a C57BL/6 or BALB/c mouse skin graft. Using the whole transcriptome sequencing-based serial analysis of gene expression (SAGE) technology, we identified 97 increasingly and 88 decreasingly expressed genes that may play important roles in allograft rejection and tolerance. Functional classification of these genes shows that apoptosis, transcription regulation, cell growth and maintenance, and signal transduction are among the frequently changed functional groups. This study provides a genome-wide view for the candidate genes of CD4+ T cells related to allotransplantation, and this report is a good resource for further microarray studies and for identifying the specific markers that are associated with clinical organ transplantations.

Key words: Allograft rejection; Gene expression profile; Genomics; Serial analysis of gene expression (SAGE); CD4+ T cells; SCID mouse

1These authors provided equal contribution to this study.
Address correspondence to Guihua Hou, M.D., Key Laboratory for Experimental Teratology of the Ministry of Education and Institute of Experimental Nuclear Medicine, School of Medicine, Shandong University, 44# Wenhua Xi Road, Jinan, Shandong 250012, P.R. China. Tel: 086-531-88382096; Fax: 086-531-88382502; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it