Cell Transplantation 23(1) Abstracts

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Cell Transplantation, Vol. 23, pp. 1-11, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X659925
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

In Search of the Pivot Point of Mechanotransduction: Mechanosensing of Stem Cells

Yi-Shiuan Liu* and Oscar K. Lee*†‡

*Stem Cell Research Center, National Yang-Ming University, Taipei, Taiwan
†Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
‡Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan

Stem cells are undifferentiated cells with the ability to self-renew and to differentiate into diverse specialized cell types; hence, they have great potential in tissue engineering and cell therapies. In addition to biochemical regulation, the physical properties of the microenvironments, such as scaffold topography, substrate stiffness, and mechanical forces, including fluid shear stress, compression, and tensile strain, can also regulate the proliferation and differentiation of stem cells. Upon physical stimuli, cytoskeleton rearrangements are expected to counterbalance the extracellular mechanical forces, trigger signaling cascades, and eventually cause epigenetic modifications. This article mainly focuses on the mechanosensing, which is the upstream event of stem cell mechanotransduction and the downstream one of physical stimuli. Putative mechanosensors such as ion channels, integrins, and cell membrane as well as primary cilia are discussed. Because mechanical environment is an important stem cell niche, identification of mechanosensors not only can elucidate the mechanisms of mechanotransduction and fate commitments but also bring new prospects of the mechanical control as well as drug development for clinical application.

Key words: Mesenchymal stem cells (MSCs); Mechanotransduction; Mechanosensor; TRP ion channels; Primary cilia; Calcium signaling

Received February 14, 2012; final acceptance November 21, 2012. Online prepub date: January 2, 2013.
Address correspondence to Oscar K. Lee, M.D., Ph.D., Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei, Taiwan. Tel: +886-2-28757391; Fax: +886-2-28757409; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 13-25, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X661337
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Intramuscular Transplantation of Myogenic Cells in Primates: Importance of Needle Size, Cell Number, and Injection Volume

Daniel Skuk, Marlyne Goulet, and Jacques P. Tremblay

Neurosciences Division-Human Genetics, CHUQ Research Center-CHUL, Quebec, QC, Canada

The aim of this study was to quantitatively define the main measurable technical parameters for the intramuscular transplantation of myogenic cells in primates. Myoblasts transduced with the gene coding for b-galactosidase were injected into the skeletal muscles of 15 monkeys. The following parameters were studied: needle size, number of cells per injection, and volume of cell suspension per injection. Monkeys were immunosuppressed with tacrolimus. The cell-injected sites were biopsied 1 or 2 months later. Biopsies were examined histologically to assess the myoblast engraftment and the muscle structure. The conclusions were as follows: (1) Needles should be thin enough to avoid important tissue damage and allow muscle regeneration as satisfactory as possible. Among those tested, 27G should be the choice if the length is consistent with depth of injection. (2) At least 100,000 cells should be delivered per centimeter of needle trajectory. (3) The smallest volumes of cell suspension per injection should be used. In this study, 1 μl/cm of injection trajectory was sufficient. In principle, these parameters apply to muscles in which no damage occurred other than the injections.

Key words: Intramuscular injection; Myoblast transplantation; Nonhuman primates; Skeletal muscle; Technical parameters

Received July 12, 2012; final acceptance December 12, 2012. Online prepub date: January 2, 2013.
Address correspondence to Daniel Skuk, M.D., Unité de recherche en Génétique humaine, Centre Hospitalier de l’Université Laval, 2705 boulevard Laurier, Québec (QC), G1V 4G2, Canada. Tel.: +1 (418) 654-2186; Fax.: +1 (418) 654-2207; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 27-38, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X662453
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Increased Reprogramming of Human Fetal Hepatocytes Compared With Adult Hepatocytes in Feeder-Free Conditions

Marc C. Hansel,*† Roberto Gramignoli,‡ William Blake,§ Julio Davila,§ Kristen Skvorak,* Kenneth Dorko,* Veysel Tahan,* Brian R. Lee,* Edgar Tafaleng,¶ Jorge Guzman-Lepe,¶ Alejandro Soto-Gutierrez,*¶ Ira J. Fox,†¶ and Stephen C. Strom†‡

*Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
†McGowan Institute for Regenerative Medicine, Pittsburgh, PA, USA
‡Department of Laboratory Medicine, Division of Pathology, Karolinska Institutet, Stockholm, Sweden
§Genetically Modified Models Center of Emphasis, Pfizer, Groton, CT, USA
¶Center for Innovative Regenerative Therapies, Department of Surgery, Transplantation Section, Children’s Hospital of Pittsburgh, Pittsburgh, PA, USA

Hepatocyte transplantation has been used to treat liver disease. The availability of cells for these procedures is quite limited. Human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) may be a useful source of hepatocytes for basic research and transplantation if efficient and effective differentiation protocols were developed and problems with tumorigenicity could be overcome. Recent evidence suggests that the cell of origin may affect hiPSC differentiation. Thus, hiPSCs generated from hepatocytes may differentiate back to hepatocytes more efficiently than hiPSCs from other cell types. We examined the efficiency of reprogramming adult and fetal human hepatocytes. The present studies report the generation of 40 hiPSC lines from primary human hepatocytes under feeder-free conditions. Of these, 37 hiPSC lines were generated from fetal hepatocytes, 2 hiPSC lines from normal hepatocytes, and 1 hiPSC line from hepatocytes of a patient with Crigler–Najjar syndrome, type 1. All lines were confirmed reprogrammed and expressed markers of pluripotency by gene expression, flow cytometry, immunocytochemistry, and teratoma formation. Fetal hepatocytes were reprogrammed at a frequency over 50-fold higher than adult hepatocytes. Adult hepatocytes were only reprogrammed with six factors, while fetal hepatocytes could be reprogrammed with three (OCT4, SOX2, NANOG) or four factors (OCT4, SOX2, NANOG, LIN28 or OCT4, SOX2, KLF4, C-MYC). The increased reprogramming efficiency of fetal cells was not due to increased transduction efficiency or vector toxicity. These studies confirm that hiPSCs can be generated from adult and fetal hepatocytes including those with genetic diseases. Fetal hepatocytes reprogram much more efficiently than adult hepatocytes, although both could serve as useful sources of hiPSC-derived hepatocytes for basic research or transplantation.

Key words: Metabolic liver disease; Reprogramming efficiency; Induced pluripotent stem cells (iPSCs); Fetal hepatocytes

Received July 28, 2012; final acceptance December 12, 2012. Online prepub date: February 4, 2013.
Address correspondence to Stephen C. Strom, Department of Laboratory Medicine, Division of Pathology, Karolinska University Hospital, F56, 141-86 Stockholm, Sweden. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 39-49, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X659844
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Preclinical Safety Studies on Autologous Cultured Human Skin Fibroblast Transplantation

Wei Zeng,*1 Shuying Zhang,*1 Dai Liu,* Mi Chai,* Jiaqi Wang,† and Yuming Zhao*

*Department of the Plastic and Reconstructive Surgery, Clinical Division of Surgery, Chinese People’s Liberation Army (PLA) General Hospital, Beijing, China
†Department of the Aesthetic and Plastic Surgery on the Face and Neck, the Plastic Surgery Hospital, Beijing, China

Recently, FDA approved the clinical use of autologous fibroblasts (LAVIVTM) for the improvement of nasolabial fold wrinkles in adults. The use of autologous fibroblasts for the augmentation of dermal and subcutaneous defects represents a potentially exciting natural alternative to the use of other filler materials for its long-term corrective ability and absence of allergic adverse effects proved by clinical application. However, compared to the clinical evidence, preclinical studies are far from enough. In this study, human skin-derived fibroblasts were cultured and expanded for both in vitro and in vivo observations. In vitro, the subcultured fibroblasts were divided into two groups. One set of cells underwent cell cycle and karyotype analysis at passages 5 and 10. The second group of cells was cocultured in medium with different concentrations of human skin extract D for the measurement of collagen concentration and cell count. In vivo, the subcultured fibroblasts were injected into nude mice subcutaneously. Biopsies were taken for morphology observation and specific collagen staining at 1, 2, and 3 months after injection. The results in vitro showed no significant differences in cell cycle distribution between passages 5 and 10. Cell proliferation and secretion were inhibited as the concentration of extract D increased. In vivo, the fibroblasts were remarkably denser on the experimental side with no dysplastic cells. Mitotic cells were easily observed at the end of the first month but were rare at the end of the third month. Type III collagen was detected at the end of the first month, while collagen type I was positive at the end of the second month. The content of both collagens increased as time passed. The above results indicated that the use of the autologous fibroblasts was safe, providing a basic support for clinical use of fibroblasts.

Key words Autologous; Fibroblasts; Transplantation; Preclinical study

Received February 19, 2012; final acceptance November 15, 2012. Online prepub date: December 4, 2012.
1These authors provided equal contribution to this study.
Address correspondence to Yuming Zhao, M.D., Department of the Plastic and Reconstructive Surgery, Chinese People’s Liberation Army (PLA) General Hospital, 28 Fuxing Road, Beijing, 100853, China. Tel: +86-10-66938150; Fax: +86-10-66938050; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 51-58, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658962
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

B-Cell Depletion Improves Islet Allograft Survival With Anti-CD45RB

Kang Mi Lee,* Heidi Yeh,* Gaoping Zhao,*† Lingling Wei,*† Matthew O’Connor,* Ryan T. Stott,* Julie Soohoo,* Kyri Dunussi,‡§ Paolo Fiorina,‡¶ Shaoping Deng,*† James F. Markmann,* and James I. Kim*

*Transplantation Unit, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
†Department of Surgery, Sichuan Provincial People’s Hospital and Sichuan Academy of Medical Sciences, Chengdu, Sichuan Province, China
‡Transplantation Research Center, Children’s Hospital of Boston, Harvard Medical School, Boston, MA, USA
§Pfizer Research, Cambridge, MA, USA
¶San Raffaele Scientific Institute, Medicine, Milan, Italy

A short course of anti-CD45RB leads to long-term islet allograft survival and donor-specific tolerance in approximately half of immunocompetent mice. We have previously demonstrated that anti-CD45RB antibody-mediated tolerance requires B-cells for cardiac allograft survival. We therefore asked whether B-cells were also required for anti-CD45RB antibody-mediated survival of islets. Unexpectedly, we found that nearly 100% of islet allografts survive long term in B-cell-deficient mice. Similarly, B-cell depletion by anti-CD22/cal augmented anti-CD45RB-mediated tolerance when administered pretransplant, although it had no effect on tolerance induction when administered posttransplant. Our results demonstrate that the role of B-cells in promoting tolerance with anti-CD45RB is graft specific, promoting tolerance in cardiac grafts but resisting tolerance in islet transplantation. These findings may help elucidate the varied action of B-cells in promoting tolerance versus rejection.

Key words: B-cells; Anti-CD45RB; Anti-CD22; Islet transplantation; Tolerance

Received June 18, 2012; final acceptance November 9, 2012. Online prepub date: November 27, 2012.
Address correspondence to James Markmann, M.D., Ph.D., 55 Fruit Street – 5 White, Massachusetts General Hospital, Boston, MA 02114, USA. Tel: +1-617-643-4533; Fax: +1-617-643-4579; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 59-72, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X659880
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Pancreatic Islet Basement Membrane Loss and Remodeling After Mouse Islet Isolation and Transplantation: Impact for Allograft Rejection

H. F. Irving-Rodgers,* F. J. Choong,† K. Hummitzsch,‡ C. R. Parish,† R. J. Rodgers,‡ and C. J. Simeonovic†

*Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland, Australia
†Department of Immunology, The John Curtin School of Medical Research, The Australian National University, Canberra, Australian Capital Territory, Australia
‡Research Centre for Reproductive Health, Discipline of Obstetrics and Gynaecology, Robinson Institute, The University of Adelaide, Adelaide, South Australia, Australia

The isolation of islets by collagenase digestion can cause damage and impact the efficiency of islet engraftment and function. In this study, we assessed the basement membranes (BMs) of mouse pancreatic islets as a molecular biomarker for islet integrity, damage after isolation, and islet repair in vitro as well as in the absence or presence of an immune response after transplantation. Immunofluorescence staining of BM matrix proteins and the endothelial cell marker platelet endothelial cell adhesion molecule-1 (PECAM-1) was performed on pancreatic islets in situ, isolated islets, islets cultured for 4 days, and islet grafts at 3–10 days posttransplantation. Flow cytometry was used to investigate the expression of BM matrix proteins in isolated islet β-cells. The islet BM, consisting of collagen type IV and components of Engelbreth–Holm–Swarm (EHS) tumor laminin 111, laminin α2, nidogen-2, and perlecan in pancreatic islets in situ, was completely lost during islet isolation. It was not reestablished during culture for 4 days. Peri- and intraislet BM restoration was identified after islet isotransplantation and coincided with the migration pattern of PECAM-1+ vascular endothelial cells (VECs). After islet allotransplantation, the restoration of VEC-derived peri-islet BMs was initiated but did not lead to the formation of the intraislet vasculature. Instead, an abnormally enlarged peri-islet vasculature developed, coinciding with islet allograft rejection. The islet BM is a sensitive biomarker of islet damage resulting from enzymatic isolation and of islet repair after transplantation. After transplantation, remodeling of both peri- and intraislet BMs restores β-cell–matrix attachment, a recognized requirement for β-cell survival, for isografts but not for allografts. Preventing isolation-induced islet BM damage would be expected to preserve the intrinsic barrier function of islet BMs, thereby influencing both the effector mechanisms required for allograft rejection and the antirejection strategies needed for allograft survival.

Key words: Islet; Basement membrane (BM); Vascular endothelial cells (VECs); Transplantation; Isograft; Allograft; Rejection

Received June 7, 2012; final acceptance November 20, 2012. Online prepub date: December 4, 2012.
Address correspondence to Dr. Charmaine J. Simeonovic, Department of Immunology, The John Curtin School of Medical Research, The Australian National University, Canberra, ACT 2601, Australia. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 73-85, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658971
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Myeloid-Derived Suppressor Cells Are Generated During Retroviral Transduction of Murine Bone Marrow

Alba Gomez,*1 Carmen Espejo,†1 Herena Eixarch,† Silvia Casacuberta-Serra,* Maria Jose Mansilla,* Rebeca Sanchez,* Sonia Pereira,* Sergio Lopez-Estevez,* Ramon Gimeno,* Xavier Montalban,† and Jordi Barquinero*

*Gene and Cell Therapy Laboratory, Vall d’Hebron Institut de Recerca (VHIR), Universitat Autònoma de Barcelona, Barcelona, Spain
†Unitat de Neuroimmunologia Clínica, Centre d’Esclerosi Múltiple de Catalunya (CEM-Cat), Vall d’Hebron Institut de Recerca (VHIR), Hospital Universitari Vall d’Hebron, Universitat Autònoma de Barcelona, Barcelona, Spain

Previous work by our group showed that transferring bone marrow cells transduced with an autoantigen into nonmyeloablated mice with experimental autoimmune encephalomyelitis induced immune tolerance and improved symptoms of the disease. Because this effect occurred in the absence of molecular chimerism, we hypothesized that the cells responsible did not have repopulating ability and that they were not mediating central but peripheral tolerance mechanisms. In the present study, we analyzed the immunophenotype of the cells that are generated in the transduction cultures and we evaluated the immunosuppressive activity of the main cell subpopulations produced. We show that both granulocytic (CD11b+ Gr-1hi) and monocytic (CD11b+ Gr-1lo) myeloid-derived suppressor cells (G- and M-MDSCs, respectively) are generated during standard 4-day γ-retroviral transduction cultures (representing about 25% and 40% of the total cell output, respectively) and that the effectively transduced cells largely consist of these two cell types. A third cell population representing about 15% of the transduced cells did not express CD45 or hematopoietic lineage markers and expressed mesenchymal stromal cell markers. Transduced total bone marrow cells and sorted M-MDSCs expressed arginase and inducible nitric oxide synthase activities, produced reactive oxygen species, and inhibited antigen-induced T-cell proliferation in vitro. Transgene-expressing MDSCs could be exploited therapeutically to induce tolerance in autoimmune diseases and in gene therapy protocols.

Key words: Myeloid-derived suppressor cells (MDSCs); Hematopoietic cells; Bone marrow cultures; Retroviral transduction; Experimental autoimmune encephalomyelitis; Autoimmune diseases

Received August 7, 2012; final acceptance November 7, 2012. Online prepub date: November 27, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Jordi Barquinero, Laboratory of Gene and Cell Therapy, Vall d’Hebron Institut de Recerca, Passeig Vall d’Hebron 119-129, Barcelona 08035, Spain. Tel: +34 932746726; Fax: +34 932746727; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 87-96, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658836
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Long-Term Functional Benefits of Epicardial Patches as Cell Carriers

Hadhami Hamdi,* Valérie Planat-Benard,† Alain Bel,*‡ Hany Neamatalla,*‡ Laetitia Saccenti,* Damelys Calderon,* Valérie Bellamy,* Martin Bon,* Marie-Cécile Perrier,§ Chantal Mandet,‡ Patrick Bruneval,‡ Louis Casteilla,† Albert A. Hagège,*‡ Michel Pucéat,* Onnik Agbulut,#1 and Philippe Menasché*‡1

*INSERM U633, Laboratory of Biosurgical Research, Paris, France
†UMR 5273 UPS, CNRS, EFS, Inserm U1031, STROMALab, Toulouse, France
‡Assistance Publique-Hôpitaux de Paris, Hôpital Européen Georges Pompidou, Department of Cardiovascular Surgery; University Paris Descartes, Sorbonne Paris Cité, Paris, France
§INSERM U970, Paris Cardiovascular Research Center, Paris, France
#Université Paris Diderot, Sorbonne Paris Cité, CNRS EAC4413, Unit of Functional and Adaptive Biology, Paris, France

Both enzymatic dissociation of cells prior to needle-based injections and poor vascularization of myocardial infarct areas are two important contributors to cell death and impede the efficacy of cardiac cell therapy. Because these limitations could be overcome by scaffolds ensuring cell cohesiveness and codelivery of angiogenic cells, we used a chronic rat model of myocardial infarction to assess the long-term (6 months) effects of the epicardial delivery of a composite collagen-based patch harboring both cardiomyogenesis-targeted human embryonic SSEA-1+ (stem cell-derived stage-specific embryonic antigen-1 positive) cardiovascular progenitors and autologous (rat) adipose tissue-derived angiogenesis-targeted stromal cells (n = 27). Cell-free patches served as controls (n = 28). Serial follow-up echocardiographic measurements of left ventricular ejection fraction (LVEF) showed that the composite patch group yielded a significantly better preservation of left ventricular function that was sustained over time as compared with controls, and this pattern persisted when the assessment was restricted to the subgroup of rats with initial LVEFs below 50%. The composite patch group was also associated with significantly less fibrosis and more vessels in the infarct area. However, although human progenitors expressing cardiac markers were present in the patches before implantation, none of them could be subsequently identified in the grafted tissue. These data confirm the efficacy of epicardial scaffolds as cell carriers for ensuring long-term functional benefits and suggest that these effects are likely related to paracrine effects and call for optimizing cross-talks between codelivered cell populations to achieve the ultimate goal of myocardial regeneration.

Key words: Cardiac patches; Embryonic stem cells (ESCs); Adipose-derived stromal cells (ADSCs); Cardiovascular progenitors; Stage-specific embryonic antigen-1 positive (SSEA-1+) cells, Cell therapy

Received March 25, 2012; final acceptance October 19, 2012. Online prepub date: November 1, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Philippe Menasché, Department of Cardiovascular Surgery, Hôpital Européen Georges Pompidou, 20, rue Leblanc, 75015 Paris, France. Tel: +33-1-5609-3622; Fax: +33-1-5609-3261; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Dr. Onnik Agbulut, Université Paris Diderot Sorbonne Paris Cité, CNRS EAC4413, Unit of Functional and Adaptive Biology, 4, rue Marie Andrée Lagroua Weill-Hallé (case 7006), 75013 Paris, France. Tel: +33-1-5727-7966; Fax: +33-1-5727-7966; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 97-110, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658845
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Improvement in Nerve Regeneration Through a Decellularized Nerve Graft by Supplementation With Bone Marrow Stromal Cells in Fibrin

Zhe Zhao,*†1 Yu Wang,*1 Jiang Peng,* Zhiwu Ren,* Li Zhang,* Quanyi Guo,* Wenjing Xu,* and Shibi Lu*

*Orthopedics Research Institute of Chinese PLA, General Hospital of Chinese PLA, Beijing, P.R. China
†Department of Orthopaedic Surgery, General Hospital of Chinese People’s Army Police Force, Beijing, P.R. China

Acellular nerve grafting is often inferior as well as an inadequate alternative to autografting for the repair of long gaps in peripheral nerves. Moreover, the injection method is not perfect. During the injection of cells, the syringe can destroy the acellular nerve structure and the limited accumulation of seed cells. To resolve this problem, we constructed a nerve graft by acellular nerve grafting. Bone marrow-mesenchymal stromal cells (BM-MSCs) were affixed with fibrin glue and injected inside or around the graft, which was then used to repair a 15-mm nerve defect in rats. The acellular nerve graft maintained its structure and composition, and its tensile strength was decreased, as determined by two-photon microscopy and a tensile testing device. In vitro, MSCs embedded in fibrin glue survived and secreted growth factors such as nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). We repaired 15-mm Sprague–Dawley rat sciatic nerve defects using this nerve graft construction, and MSCs injected around the graft helped improve nerve regeneration and functional recovery of peripheral nerve lesions as determined by functional analysis and histology. Therefore, we conclude that supplying MSCs in fibrin glue around acellular nerves is successful in maintaining the nerve structure and can support nerve regeneration similar to the direct injection of MSCs into the acellular nerve for long nerve defects but may avoid destroying the nerve graft. The technique is simple and is another option for stem cell transplantation.

Key words: Acellular nerve; Cell encapsulation; Mesenchymal stromal cells; Fibrin glue; Nerve regeneration

Received December 7, 2011; final acceptance October 23, 2012. Online prepub date: November 1, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Jiang Peng, Orthopedics Research Institute of Chinese PLA, General Hospital of Chinese PLA, Fuxing Road 28, Haidian District, Beijing 100853, P.R. China. Tel: +86-10-6693-9205; Fax: +86-10-6816-9305; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 111-132, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X658944
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Anticonvulsant Effects by Bilateral and Unilateral Transplantation of GABA-Producing Cells Into the Subthalamic Nucleus in an Acute Seizure Model

Annelie Handreck,*† Bianca Backofen-Wehrhahn,* Sonja Bröer,*† Wolfgang Löscher,*† and Manuela Gernert*†

*Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine, Hannover, Germany
†Center for Systems Neuroscience, Hannover, Germany

Neural transplantation of GABA-producing cells into key structures within seizure-suppressing circuits holds promise for medication-resistant epilepsy patients not eligible for resection of the epileptic focus. The substantia nigra pars reticulata (SNr), a basal ganglia output structure, is well known to modulate different seizure types. A recent microinjection study by our group indicated that the subthalamic nucleus (STN), which critically regulates nigral activity, might be a more promising target for focal therapy in epilepsies than the SNr. As a proof of principle, we therefore assessed the anticonvulsant efficacy of bilateral and unilateral allografting of GABA-producing cell lines into the STN using the timed intravenous pentylenetetrazole seizure threshold test, which allows repeated seizure threshold determinations in individual rats. We observed (a) that grafted cells survived up to the end of the experiments, (b) that anticonvulsant effects can be induced by bilateral transplantation into the STN using immortalized GABAergic cells derived from the rat embryonic striatum and cells additionally transfected to obtain higher GABA synthesis than the parent cell line, and (c) that anticonvulsant effects were observed even after unilateral transplantation into the STN. Neither grafting of control cells nor transplantation outside the STN induced anticonvulsant effects, emphasizing the site and cell specificity of the observed anticonvulsant effects. To our knowledge, the present study is the first showing anticonvulsant effects by grafting of GABA-producing cells into the STN. The STN can be considered a highly promising target region for modulation of seizure circuits and, moreover, has the advantage of being clinically established for functional neurosurgery.

Key words: Substantia nigra pars reticulata (SNr); Basal ganglia; Epilepsy; Glutamic acid decarboxylase (GAD); Pentylenetetrazole

Received December 16, 2011; final acceptance October 27, 2012. Online prepub date: November 27, 2012.
Address correspondence to Prof. Dr. Manuela Gernert, Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine Hannover, Bünteweg 17, D-30559 Hannover, Germany. Tel: +49-511-953-8527; Fax: +49-511-953-8581; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 133-138, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X659835
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Comparative Outcomes of Penetrating and Component Endothelial Cell Corneal Allografts in Outbred Sheep

Richard A. D. Mills,*†1 Sonja Klebe,†1 Douglas J. Coster,* and Keryn A. Williams*

*Department of Ophthalmology, Flinders University, Adelaide, South Australia, Australia
†Department of Anatomical Pathology, Flinders University, Adelaide, South Australia, Australia

Lamellar (component cell) corneal transplantation is replacing penetrating keratoplasty for some corneal disorders in humans, but the relative risks of immunological graft rejection for the two procedures remain uncertain. A model of component endothelial cell keratoplasty (endokeratoplasty) was developed in outbred sheep. Clinical and histological graft outcomes after endokeratoplasty were then compared with contemporaneous penetrating corneal allografts. No topical or systemic immunosuppression was administered to any recipient sheep. Endothelial cell allografts (n = 10) took significantly longer to achieve perfect transparency following surgery than did penetrating corneal grafts (n = 7) (day 10 vs. day 4; p = 0.003; two-tailed Mann–Whitney U test). The median day to rejection of penetrating grafts was postoperative day 18; for endothelial cell grafts, it was day 48 (p = 0.04; two-tailed Mann–Whitney U test). The clinical courses of the two procedures were therefore quite different. Penetrating grafts gained clarity quickly but exhibited rapid graft neovascularization. Clinical rejection was preceded by inflammation in the anterior segment. Endothelial cell grafts exhibited a fluctuating, more indolent course of opacification, although all did eventually fail. Histological analysis confirmed immunological rejection in all failed grafts, but with different patterns of leukocytic infiltration in endokeratoplasties compared with penetrating keratoplasties. Inflammatory cells in endothelial cell grafts were generally fewer in number and were more often found in the posterior stroma. We conclude that, in the absence of immunosuppression, all endothelial cell allografts do undergo immunological rejection, albeit at a slower rate than penetrating grafts.

Key words: Corneal transplantation; Sheep; Endothelial cell keratoplasty (Endokeratoplasty); Immunological rejection

Received August 21, 2012; final acceptance November 15, 2012. Online prepub date: December 4, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Keryn Williams, Department of Ophthalmology, Flinders University, Flinders Medical Centre, Bedford Park, SA 5042, Australia. Tel: +61-88-204-5047; Fax: +61-88-277-0899; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it