Cell Transplantation 20(10) Abstracts

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Cell Transplantation, Vol. 20, pp. 1499–1514, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X557281
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved


A High-Fat/High-Cholesterol Diet Inhibits Growth of Fetal Hippocampal Transplants via Increased Inflammation

L. R. Freeman,* B. J. Small,† P. C. Bickford,‡§ C. Umphlet,* and A.-Ch. Granholm*

*Department of Neurosciences and the Center on Aging, Medical University of South Carolina, Charleston, SC, USA
†School of Aging Studies, University of South Florida, Tampa, FL, USA
‡Department of Neurosurgery, Center for Excellence for Aging and Brain Repair, University of South Florida, Tampa, FL, USA
§James A Haley Veterans’ Hospital Medical Center, Tampa, FL, USA

A diet containing high levels of saturated fat and cholesterol is detrimental to many aspects of health and is known to lead to obesity, metabolic syndrome, heart disease, diabetes, and cancer. However, the effects of a diet rich in saturated fat and cholesterol on the brain are not currently well understood. In order to determine direct effects of a high saturated fat and cholesterol diet upon fetal hippocampal tissue, we transplanted hippocampal grafts from embryonic day 18 rats to the anterior eye chamber of 16-month-old host animals that were fed either a normal rat chow diet or a 10% hydrogenated coconut oil +  2% cholesterol diet (HFHC diet) for 8 weeks. One eye per rat received topical application of an IL-1 receptor antagonist (IL-1Ra, Kineret®) and the other served as a saline control. Results revealed that the HFHC diet led to a marked reduction in hippocampal transplant growth, and detrimental effects of the diet were alleviated by the IL-1 receptor antagonist IL-1Ra. Graft morphology demonstrated that the HFHC diet reduced organotypical development of the hippocampal neuronal cell layers, which was also alleviated by IL-1Ra. Finally, grafts were evaluated with markers for glucose transporter expression, astrocytes, and activated microglia. Our results demonstrate significant effects of the HFHC diet on hippocampal morphology, including elevated microglial activation and reduced neuronal development. IL-1Ra largely blocked the detrimental effects of this diet, suggesting a potential use for this agent in neurological disorders involving neuroinflammation.

Key words: Cholesterol; Saturated fat; Transplantation; Hippocampus; Inflammation

Address correspondence to Linnea Freeman at her current address: Department of Aging and Neurodegeneration, Pennington Biomedical Research Center, 6400 Perkins Road, Baton Rouge, LA 70808, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1515–1527, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X547435
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Multitract Microtransplantation Increases the Yield of DARPP-32-Positive Embryonic Striatal Cells in a Rodent Model of Huntington’s Disease

Wei Jiang,*†1 Fabian Büchele,*1 Anna Papazoglou,* Máté Döbrössy,* and Guido Nikkhah*

*Laboratory of Molecular Neurosurgery, Department of Stereotactic and Functional Neurosurgery, Neurocentre, Albert-Ludwigs-University of Freiburg, Freiburg, Germany
†Department of Neurosurgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science & Technology, Wuhan, China

Embryonic striatal graft-mediated functional recovery in the rodent lesion model of Huntington’s disease (HD) has been shown to correlate with the proportion of dopamine- and adenosine 3′,5′-monophosphateregulated phosphoprotein with a molecular weight of 32 kDa (DARPP-32)-positive neurons in the graft. The current study investigated the impact of graft distribution on the yield of DARPP-32-positive cells in the grafts following either single-tract or multitract cell delivery protocols using the microtransplantation approach. Cells derived from the whole ganglionic eminence of E15 rat embryos, ubiquitously expressing green fluorescent protein (GFP), were implanted into unilaterally QA-lesioned rat striatum either as 2 × 1.8 μl macrodeposits in a single tract, or as 18 × 0.2 μl microdeposits disseminated over six needle, multitract, penetrations. For both groups, an ultrathin glass capillary with an outer diameter of 50 μm was used. Histological assessment at 4 months after transplantation showed nearly twofold increase of DARRP-32-positive striatal-like neurons in the multitract compared to the single-tract group. However, the cellular make-up of the grafts did not translate into functional differences as tested in a basic spontaneous behavior test. Furthermore, the volumetric values for overall volume, DARPP-32-positive patches, and dopaminergic projection zones were similar between both groups. The results show that distribution of fetal striatal tissue in multiple submicroliter deposits provides for an increased yield of striatal-like neurons, potentially due to the enlargement of the graft–host border area intensifying the graft’s exposure to host-derived factors. Furthermore, the use of embryonic tissue from GFP donors was validated in cell-based therapy studies in the HD model.

Key words: Microtransplantation; Multitract; Embryonic striatal neurons; Huntington’s disease model; Green fluorescent protein (GFP)

1These authors provided equal contribution to this work.

Address correspondence to Máté Döbrössy, Laboratory of Molecular Neurosurgery, Department of Stereotactic and Functional Neurosurgery, Neurocentre, Albert-Ludwigs-University of Freiburg, Breisacher Str. 64, 79106 Freiburg, Germany. Tel: 49-761-270-5036; Fax: 49-761-270-9303; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1529–1545, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564067
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Efficient Derivation and Concise Gene Expression Profiling of Human Embryonic Stem Cell-Derived Mesenchymal Progenitors (EMPs)

Men-Luh Yen,* Chun-Han Hou,† Kai-Yen Peng,* Pei-Chi Tseng,* Shih-Sheng Jiang,‡ Chia-Tung Shun,§ Yao-Chang Chen,§ and Min-Liang Kuo¶

*Department of Primary Care Medicine & Department of Obstetrics/Gynecology, National Taiwan University Hospital & College of Medicine, National Taiwan University, Taipei, Taiwan
†Department of Orthopedic Surgery, National Taiwan University Hospital, Taipei, Taiwan
‡National Institute of Cancer Research, National Health Research Institutes, Zhunan, Taiwan
§Department of Pathology and Institute of Forensic Medicine, National Taiwan University Hospital & College of Medicine, National Taiwan University, Taipei, Taiwan
¶Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan

New potential sources of stem cells for clinical application include bone marrow mesenchymal stem cells (BMMSCs), human embryonic stem cells (hESCs), and induced pluripotent stem cells (iPS). However, each source is not without its own concerns. While research continues in an effort to overcome these problems, the generation of mesenchymal progenitors from existing hESC lines may circumvent many of these issues. We report here a simple and efficient method of generating hESC-derived mesenchymal progenitors (EMPs) and transcriptome profiling using a concise, custom-designed, oligomnucleotide gene expression microarray. Characterization of EMPs shows that these cells are similar to BMMSCs in terms of differentiation capacity as well as cell surface marker expression. In addition, EMPs express several ESC markers and HLA-G, a nonclassical MHC class I molecule with immunomodulatory properties. Morevoer, EMPs possess significantly enhanced proliferative ability over BMMSCs during which karyotypic stability was maintained. Although derived from hESCs, EMPs do not form any tumors in immunocompromised mice. To efficiently profile gene expression in multiple samples, we designed an oligoarray to probe just over 11,000 genes highly expressed in stem cells. We found that the transcriptome of EMPs is more similar to BMMSCs than hESCs. Both cell types highly express genes involved in processes related to the cytoskeleton, extracellular matrix, and cell adhesion, but EMPs show higher expression of genes involved in cell proliferation whereas BMMSCs showed higher expression of immune-related genes. Based on our data, EMPs may be an accessible source of mesenchymal progenitor for therapeutic use.

Key words: Embryonic stem cells; Bone marrow mesenchymal stem cells (BMMSCs); Transcriptome profiling; Ontology analysis

Address correspondence to Men-Luh Yen, Department of Primary Care & Department of Obstetrics/Gynecology, National Taiwan University Hospital & College of Medicine, National Taiwan University, No. 1, Section 1, Jen-Ai Road, 100, Taipei, Taiwan. Tel: +886-2-23123456, ext. 71560; Fax: +886-2-23911302; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Min-Liang Kuo, Ph.D., Institute of Toxicology, College of Medicine, National Taiwan Univeristy, No. 1, Section 1, Jen-Ai Road, 100, Taipei, Taiwan. Tel: +886-2-23123456, ext. 88607; Fax: +886-2-23410217; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1547–1559564076, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Mesenchymal Stem Cells Are Susceptible to Lysis by CD8+ T Cells and NK Cells

Meindert J. Crop,* Sander S. Korevaar,* Ronella de Kuiper,* Jan N. M. IJzermans,† Nicole M. van Besouw,* Carla C. Baan,* Willem Weimar,* and Martin J. Hoogduijn*

*Department of Internal Medicine, Erasmus University Medical Center, Rotterdam, The Netherlands
†Department of Surgery, Erasmus University Medical Center, Rotterdam, The Netherlands

There is growing interest in the use of mesenchymal stem cells (MSCs) to improve the outcome of organ transplantation. The immunogenicity of MSCs is, however, unclear and is important for the efficacy of MSC therapy and for potential sensitization against donor antigens. We investigated the susceptibility of autologous and allogeneic MSCs for lysis by CD8+ T-lymphocytes and NK cells in a kidney transplant setting. MSCs were derived from adipose tissue of human kidney donors and were CD90+, CD105+, CD166+, and HLA class I+. They showed differentiation ability and immunosuppressive capacity. Lysis of MSCs by peripheral blood mononuclear cells (PBMCs), FACS-sorted CD8+ T cells, and NK cells was measured by europium release assay. Allogeneic MSCs were susceptible for lysis by cytotoxic CD8+ T cells and NK cells, while autologous MSCs were lysed by NK cells only. NK cell-mediated lysis was inversely correlated with the expression of HLA class I on MSCs. Lysis of autologous MSCs was not dependent on culturing of MSCs in FBS, and MSCs in suspension as well as adherent to plastic were lysed by NK cells. Pretransplant recipient PBMCs did not lyse donor MSCs, but PBMCs isolated 3, 6, and 12 months after transplantation showed increasing lysing ability. After 12 months, CD8+ T-cell-mediated lysis of donor MSCs persisted, indicating there was no evidence for desensitization against donor MSCs. Lysis of MSCs is important to take into account when MSCs are considered for clinical application. Our results suggest that the HLA background of MSCs and timing of MSC administration are important for the efficacy of MSC therapy.

Key words: Mesenchymal stem cells (MSCs); Cytotoxicity; NK cells; CD8+ T cells; Transplantation

Address correspondence to M. J. Hoogduijn, Ph.D., Department of Internal Medicine, Erasmus University Medical Center, Dr. Molewaterplein 50, Room Ee559, P.O. box 2040, 3015 GE, Rotterdam, The Netherlands. Tel: +31-(0)10-7035419; Fax: +31-(0)10-7044718; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1561–1574, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X557254
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Mesenchymal Stem Cells Restore Lung Function by Recruiting Resident and Nonresident Proteins

Philipp Jungebluth,* Mark Luedde,† Elisabet Ferrer,‡ Tom Luedde,§ Mihael Vucur,§ Victor I. Peinado,‡ Tetsuhiko Go,¶ Catharina Schreiber,# Maximilian von Richthofen,# Augustinus Bader,# Johannes Haag,# Kai H. Darsow,# Sebastian J. Bartel,# Harald A. Lange,# Dario Furlani,** Gustav Steinhoff,** and Paolo Macchiarini*

*Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Huddinge, Stockholm, Sweden
†Department of Cardiology, University of Kiel, Kiel, Germany
‡Hospital Clinic, University of Barcelona, Barcelona, Spain
§Department of Gastroenterology, University Hospital Aachen, Aachen, Germany
¶Department of General Thoracic and Breast-Endcrinological Surgery, Kagawa University Miki-Cho, Kagawa, Japan
#Biomedical-Biotechnological Center, Leipzig, Leipzig, Germany
**Department of Cardiac Surgery, University Rostock, Rostock, Germany

Because human lungs are unlikely to repair or regenerate beyond the cellular level, cell therapy has not previously been considered for chronic irreversible obstructive lung diseases. To explore whether cell therapy can restore lung function, we administered allogenic intratracheal mesenchymal stem cells (MSCs) in the trachea of rats with chronic thromboembolic pulmonary hypertension (CTEPH), a disease characterized by single or recurrent pulmonary thromboembolic obliteration and progressive pulmonary vascular remodeling. MSCs were retrieved only in high pressure-exposed lungs recruited via a homing stromal derived factor-1α/CXCR4 pathway. After MSC administration, a marked and long-lasting improvement of all clinical parameters and a significant change of the proteome level were detected. Beside a variation of liver proteome, such as caspase-3, NF-κB, collagen1A1, and α-SMA, we also identified more than 300 resident and nonresident lung proteins [e.g., myosin light chain 3 (P16409) or mitochondrial ATP synthase subunit alpha (P15999)]. These results suggest that cell therapy restores lung function and the therapeutic effects of MSCs may be related to protein-based tissue reconstituting effects.

Key words: Mesenchymal stem cells (MSCs); Protein expression; Stromal derived factor-1; Pulmonary hypertension

Address correspondence to Paolo Macchiarini, M.D., Ph.D., Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Alfred Nobels Alle´ 8, Huddinge, SE-171 77 Stockholm, Sweden. Tel: +39 0557949984; Fax: +390557947935; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1575–1588, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X557191
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Effect of Two- and Three-Dimensional Cell Culture on the Chondrogenic Potential of Human Adipose-Derived Mesenchymal Stem Cells After Subcutaneous Transplantation With an Injectable Hydrogel

Christophe Merceron,*† Sophie Portron,*† Martial Masson,*† Julie Lesoeur,*† Borhane Hakim Fellah,*†‡¶ Olivier Gauthier,*†‡ Olivier Geffroy,*†§ Pierre Weiss,*† Jérôme Guicheux,*†1 and Claire Vinatier*†¶1

*INSERM (Institut National de la Santé et de la Recherche Médicale), UMRS 791, Université de Nantes, Laboratoire d’Ingénierie Ostéo-Articulaire et Dentaire, Group STEP “Skeletal tissue Engineering and Physiopathology,” Faculté de chirurgie dentaire, Nantes Cedex 1, France
†PRES-UNAM, UFR Odontologie, Université de Nantes, Nantes, France
‡Ecole Nationale vétérinaire de Nantes (ONIRIS), Service de chirurgie expérimentale (CRIP), Atlanpôle-La Chantrerie, Nantes Cedex 3, France
§Ecole Nationale vétérinaire de Nantes (ONIRIS), Service de chirurgie équine, Atlanpôle-La Chantrerie, Nantes Cedex 3, France
¶Graftys SA-Eiffel Park-Bâtiment D-415, Aix en Provence Cedex 3, France

Articular cartilage is an avascular tissue composed of chondrocytes, a unique cell type responsible for abundant matrix synthesis and maintenance. When damaged, it never heals spontaneously under physiological circumstances. Therefore, the delivery of mesenchymal stem cells using hydrogel has been considered for cartilage repair. This study aims at investigating the influence of in vitro chondrogenic differentiation of human adipose tissue-derived stem cells (hATSCs) on in vivo cartilage formation when associated with a cellulose-based self-setting hydrogel (Si-HPMC). hATSCs were characterized for their proliferation, surface marker expression, and multipotency. The in vitro chondrogenic potential of hATSCs cultured within Si-HPMC in control or chondrogenic medium was evaluated by measuring COL2A1, ACAN, SOX9, and COMP expression by real-time PCR. Alcian blue and type II collagen staining were also performed. To determine whether in vitro chondrogenically differentiated hATSCs may give rise to cartilage in vivo, cells differentiated as a monolayer or in pellets were finally associated with Si-HPMC and implanted subcutaneously into nude mice. Cartilage formation was assessed histologically by alcian blue and type II collagen staining. Our data demonstrate that hATSCs exhibited proliferation and self-renewal. hATSCs also expressed typical stem cell surface markers and were able to differentiate towards the adipogenic, osteogenic, and chondrogenic lineages. Real-time PCR and histological analysis indicated that Si-HPMC enabled chondrogenic differentiation of hATSCs in inductive medium, as demonstrated by increased expression of chondrogenic markers. In addition, histological analysis of implants showed that chondrogenically differentiated hATSCs (monolayers or pellets) have the ability to form cartilaginous tissue, as indicated by the presence of sulphated glycosaminoglycans and type II collagen. This study therefore suggests that an in vitro induction of hATSCs in 2D was sufficient to obtain cartilaginous tissue formation in vivo. Si-HPMC associated with autologous hATSCs could thus be a significant tool for regenerative medicine in the context of cartilage damage.

Key words: Tissue engineering; Cartilage; Hydrogel; Human adipose-derived stem cells (hATSCs); Chondrogenic differentiation

1These authors provided equal contribution to this work.
Address correspondence to Prof. Jérôme Guicheux, Ph.D., INSERM U 791, Laboratoire d’ingénierie Ostéo-Articulaire et Dentaire (LIOAD), Faculté de Chirurgie Dentaire, 1 Place Alexis Ricordeau, 44042 Nantes Cedex 1, France. Tel: +33 240412919; Fax: +33 240083712; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1589–1602, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564094
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Hypoxic Conditions During Expansion Culture Prime Human Mesenchymal Stromal Precursor Cells for Chondrogenic Differentiation in Three-Dimensional Cultures

Jana Müller,* Karin Benz,* Michael Ahlers,† Christoph Gaissmaier,‡ and Jürgen Mollenhauer*‡

*NMI Natural and Medical Sciences Institute, University of Tübingen, Reutlingen, Germany
†GELITA AG, Eberbach, Germany
‡TETEC AG, Reutlingen, Germany

Cell-based approaches using mesenchymal stromal precursor cells (MSCs) for the regeneration of intervertebral discs are attracting increased interest, even though the intervertebral disc is a very demanding environment. Implanted cells eventually face acidic pH, hypoxia, and a lack of nutrients. While the regenerative potential of MSCs for skeletal tissues has been well described, it is still questionable whether human MSCs can be prepared for prolonged survival and proper functioning and whether they can differentiate under the adverse conditions encountered in the disc. Here we examined the influence of hypoxia during expansion and differentiation on the chondrogenesis of MSCs. Chondrogenic differentiation was performed in in situ solidifying gelatin hydrogels, which represent a suitable matrix for delivering and anchoring cells within the disc tissue. To consider limitations in nutrition in the intervertebral disc, differentiation was performed at low cell concentrations in the gelatin hydrogels. Standard high-density micromass cultures served as reference controls. To determine the quality of chondrogenesis we analyzed typical marker molecules such as collagen types I, II, X, Sox-9, MIA, and aggrecan mRNA using RT-qPCR and determined protein deposition by histological stainings and biochemical methods. We could demonstrate that in gelatin-based hydrogels chondrogenic differentiation of human MSCs is possible at low cell concentrations. The quality of chondrogenic differentiation could be improved by hypoxia. Best results were obtained when the entire in vitro process, including MSC expansion and subsequent differentiation, was done under hypoxic conditions. MSCs that were expanded under reduced oxygen tension were primed for a chondrogenic differentiation.

Key words: Mesenchymal stromal precursor cell; Gelatin hydrogel; Hypoxia; Intervertebral disc; Chondrogenic differentiation

Address correspondence to Jürgen Mollenhauer, TETEC AG, Aspenhaustr. 18, 72770 Reutlingen, Germany. Tel: +49-7121-5148774; Fax: +49-7121-5148761; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or NMI Natural and Medical Sciences Institute at the University of Tübingen, Markwiesenstr. 55, 72770 Reutlingen, Germany. Tel: +49-7121-5153034; Fax: +49-7121-5153016; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1603–1620, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564517
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Morphological Changes in the Ovine Carotid Artery Wall Induced by Cold Storage

Arthur Smardencas* and Ian Birchall†

*Department of Forensic Medicine, Monash University, Clayton, Victoria, Australia
†Florey Neuroscience Institutes, University of Melbourne, Victoria, Australia

Blood vessels obtained from cadavers and amputated limbs stored at 4°C (i.e., cold stored) potentially represent an economical and readily sourced alternative to autologous vessels and synthetic prostheses for vascular reconstructive surgery. However, cold-stored vessels would need to have a reduced antigenicity and an antithrombogenic autologous endothelial cell (EC) lining before they could function as patent vascular allografts. The aim of this study was to determine the effect of cold storage for 1–16 weeks on the morphology of the ovine carotid artery wall. Ovine carotid arteries (n =  6) were rinsed and flushed with 0.9% saline, cut into segments, wrapped in 0.9% saline-soaked gauze, and stored at 4°C for 1, 2, 4, 8, or 16 weeks. Following storage, the segments were sampled and the samples fixed and sectioned for light microscopic, immunohistochemical, or transmission electron microscopic examination. After 1 and 2 weeks the ECs were karyolitic or contained pyknotic nuclei. After 4 weeks the EC layer was depleted, the subendothelial matrix exposed, and the number of smooth muscle cells (SMCs) and fibroblasts reduced. The 8- and 16-week samples demonstrated complete loss of the EC lining and only occasional remnants of SMCs or fibroblasts. The subendothelial basement membrane appeared to undergo degradative changes as early as 1 week following cold storage. At each time point examined, the subendothelial connective tissue stroma, the internal elastic lamina (IEL), and the collagenous and elastic extracellular framework were retained. These results demonstrate that the ovine carotid artery wall progressively loses its cells but retains its extracellular components during cold storage for up to 16 weeks. They suggest that cold-stored vessels may function as allografts with a reduced antigenicity for vascular reconstructive surgery. It is conceivable that seeded autologous ECs may be used to restore the antithrombogenic EC lining prior to graft implantation.

Key words: Endothelium; Collagen; Elastin; Extracellular matrix; Fibroblast; Smooth muscle cell

Address correspondence to Arthur Smardencas, Florey Neuroscience Institutes, University of Melbourne, Parkville, Victoria, Australia, 3010. Tel: 0417 386 139; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1621–1628, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564049
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Cardiomyoplasty Improves Contractile Reserve After Myocardial Injury in Mice: Functional and Morphological Investigations With Reconstructive Three-Dimensional Echocardiography

Alexander Ghanem,*1 Wilhelm Röll,†1 Toktam Bostani,† Oliver Dewald,† Bernd K. Fleischmann,‡ Jörg Stypmann,§ Georg Nickenig,* and Klaus Tiemann§

*Department of Medicine—Cardiology, University of Bonn, Bonn, Germany
†Department of Cardiac Surgery, University of Bonn, Bonn, Germany
‡Institute of Physiology, University of Bonn, Bonn, Germany
§Department of Cardiology and Angiology, University Hospital of Muenster, Muenster, Germany

Cellular cardiomyoplasty (CMP) is a novel therapeutic approach to myocardial injury (MI). Post-MI remodeling of the left ventricle (LV) comprises dilatation and impairment of systolic function and gives rise to progressive hemodynamic deterioration. We aimed to investigate: a) the impact of CMP on global and regional parameters of LV remodeling (LVR) as well as contractile reserve and b) the suitability and validityof different echocardiographic methods in this scenario. Murine ventricular cardiomyocytes (E13.5–E16.5) were transplanted into cryolesioned hearts of male HIM-OF1 mice. Echocardiography was performed at rest 4 and 14 days postoperatively. For quantification of akinetic myocardial mass and contractile reserve 2 weeks postoperatively additionally low-dose dobutamine stress echocardiography was conducted. Reconstructive 3D-echocardiography (r3D-echo) was compared to “plain” echocardiographic investigations and was compared to invasive measurements with conduction catheter. CMP significantly attenuated LV dilatation and reduced LV function decline on day 14, as obtained with all echocardiographic modalities and confirmed with conduction catheter measurements. In contrast to plain echocardiography and invasive testing, r3D-echo allowed noninvasive quantification of scar size and assessment of regional contractile reserve. Cell transplanted hearts demonstrated a significant decrease of akinetic myocardial mass (−CMP: 13 ± 2%; +CMP 7 ± 1%; p < 0.001) and increased regional contractile reserve, an indirect sign of myocardial viability. The present study demonstrates beneficial effects of CMP on global and regional parameters of LVR and contractile reserve after MI. In contrast to “simple” 2D echocardiography, r3D-echo allowed the assessment of regional contractile reserve and quantification of akinetic myocardial mass as additive functional and morphological measures of LVR.

Key words: Cellular cardiomyoplasty (CMP); Myocaridal injury; Echocardiography; Left ventricular remodeling (LVR)

1These authors provided equal contribution to this work.
Address correspondence to Dr. med. Alexander Ghanem, Department of Medicine—Cardiology, University of Bonn, Bonn, Germany. Tel: +49 228 2871-5507; Fax: +49 228 2871-1631; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1629–1639, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564526
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Angiographic Demonstration of Neoangiogenesis After Intra-arterial Infusion of Autologous Bone Marrow Mononuclear Cells in Diabetic Patients With Critical Limb Ischemia

Rafael Ruiz-Salmeron,* Antonio de la Cuesta-Diaz,* Manuel Constantino-Bermejo,* Immaculada Pérez-Camacho,* Francisco Marcos-Sánchez,* Abdelkrim Hmadcha,†‡ and Bernat Soria†‡

*Hospitales Universitarios San Lázaro and Virgen Macarena, Sevilla, Spain
†Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), Sevilla, Spain
‡CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Barcelona, Spain

Critical limb ischemia in diabetic patients is associated with high rates of morbidity and mortality. Suboptimal responses to the available medical and surgical treatments are common in these patients, who also demonstrate limited vascular homeostasis. Neovasculogenesis induced by stem cell therapy could be a useful approach for these patients. Neovasculogenesis and clinical improvement were compared at baseline and at 3 and 12 months after autologous bone marrow-derived mononuclear cell (BMMNC) transplantation in diabetic patients with peripheral artery disease. We conducted a prospective study to evaluate the safety and efficacy of intra-arterial administration of autologous BMMNCs (100–400 × 106 cells) in 20 diabetic patients with severe below-the-knee arterial ischemia. Although the time course of clinical effects differed among patients, after 12 months of follow-up all patients presented a notable improvement in the Rutherford-Becker classification, the University of Texas diabetic wound scales, and the Ankle-Brachial Index in the target limb. The clinical outcome was consistent with neovasculogenesis, which was assessed at 3 months by digital subtraction angiography and quantified by MetaMorph software. Unfortunately, local cell therapy in the target limb had no beneficial effect on the high mortality rate in these patients. In diabetic patients with critical limb ischemia, intra-arterial perfusion of BMMNCs is a safe procedure that generates a significant increase in the vascular network in ischemic areas and promotes remarkable clinical improvement.

Key words: Bone marrow-derived mononuclear cells (BMMNCs); Peripheral artery disease (PAD);Critical limb ischemia (CLI); Ankle-brachial index (ABI); Digital subtraction angiography (DSA); Below the knee

Address correspondence to Bernat Soria, M.D., Ph.D., Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), Américo Vespucio s/n, 41092 Sevilla, Spain. Tel: (+34) 954468004; Fax: (+34) 954461664; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1641–1647, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564058
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Improving Efficacy of Clinical Islet Transplantation With Iodixanol-Based Islet Purification, Thymoglobulin Induction, and Blockage of IL-1β and TNF-α

Shinichi Matsumoto,* Morihito Takita,* Damien Chaussabel,† Hirofumi Noguchi,* Masayuki Shimoda,‡ Koji Sugimoto,* Takeshi Itoh,* Daisuke Chujo,† Jeff SoRelle,§ Nicholas Onaca,¶ Bashoo Naziruddin,¶ and Marlon F. Levy¶

*Baylor All Saints Islet Cell Laboratory, Fort Worth, TX, USA
†Baylor Institute for Immunology Research, Dallas, TX, USA
‡Baylor University Medical Center, Dallas, TX 75246, USA
§Institute of Biomedical Studies, Baylor University, Waco, TX, USA
¶Baylor Regional Transplant Institute, Dallas, TX, USA

Poor efficacy is one of the issues for clinical islet transplantation. Recently, we demonstrated that pancreatic ductal preservation significantly improved the success rate of islet isolation; however, two transplants were necessary to achieve insulin independence. In this study, we introduced iodixanol-based purification, thymoglobulin induction, and double blockage of IL-1β and TNF-α as well as sirolimus-free immunosuppression to improve the efficacy of clinical islet transplantation. Nine clinical-grade human pancreata were procured. Pancreatic ductal preservation was performed using ET-Kyoto solution in all cases. When the isolated islets met the clinical criteria, they were transplanted. We utilized two methods of immunosuppression and antiinflammation. The first protocol prescribed daclizumab for induction, then sirolimus and tacrolimus to maintain immunosuppression. The second protocol used thymoglobulin for induction and tacrolimus and mycophenolate mofetil to maintain immunosuppression. Eternacept and anakinra were administered as anti-inflammatory drugs. The total amount of insulin required, HbA1c, and the SUITO index were determined to analyze and compare the results of transplantation. All isolated islet preparations (9/9) met the criteria for clinical transplantation, and they were transplanted into six type 1 diabetic patients. All patients achieved insulin independence with normal HbA1c levels; however, the first protocol required two islet infusions (N = 3) and the second protocol only required a single infusion (N = 3). The average SUITO index, at 1 month after a single-donor islet transplantation, was significantly higher in the second protocol (49.6 ± 8.3 vs. 19.3 ± 6.3, p < 0.05). Pancreatic ductal preservation, iodixanol-based purification combined with thymoglobulin induction, and blockage of IL-1 β  and TNF- α  as well as sirolimus-free immunosuppression dramatically improved the efficacy of clinical islet transplantations. This protocol enabled us to perform successful single-donor islet transplantations. Further large-scale studies are necessary to confirm these results and clarify the mechanism of each component.

Key words: Islet transplantationl; Single donor; Thymoglobulin; Interleukin-1 β  (IL-1 β); Tumor necrosis factor- α  (TNF- α)

Address correspondence to Shinichi Matsumoto, M.D., Ph.D., Baylor All Saints Islet Cell Laboratory, 1400 8th Avenue, Fort Worth, TX 76104, USA. Tel: +1-817-922-2570; Fax: +1-817-922-4645; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


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

Brief Communication

Alterations of the Female Reproductive System in Islet Recipient Receiving Immunosuppression

Maria Isabel Del Olmo Garcia,*† Vincenzo Lauriola,*‡ Altagracia Gomez Aracena,*§ Shari Messinger,* Andrea Corrales,* Camillo Ricordi,*¶#** and Rodolfo Alejandro*#

*Diabetes Research Institute, Miami, FL, University of Miami Miller School of Medicine, USA
†Department of Endocrinology and Nutrition, University Hospital La Fe, Valencia, Spain
‡Facoltá di Scienze Motorie, Universitá degli Studi di Milano, Milano, Italy
§Instituto Tecnologico de Santo Domingo (INTEC), Santo Domingo, Dominican Republic
¶DeWitt Daughtry Family Department of Surgery, University of Miami Miller School of Medicine, Miami, FL, USA
#Department of Medicine, University of Miami Miller School of Medicine, Miami, FL, USA
**Jackson Memorial Hospital-University of Miami Transplant Institute, University of Miami Miller School of Medicine, Miami, FL, USA

Pancreatic islet allotransplantation is an option for patients with unstable type 1 diabetes mellitus (T1DM). Major improvements in islet isolation techniques and the implementation of steroid-free immunosuppressive regimens can maintain insulin independence in the majority of T1DM for at least 1 year after transplantation. Recent studies have emphasized the impact of sirolimus on female reproductive tract. In this communication we report on the alterations of the female reproductive tract in 18 chronically immunosuppressed patients with T1DM following allogenic islet transplantation. Previous research has shown development of ovarian cysts in islet transplant patients receiving sirolimus. We extensively reevaluated this and other possible side effects on the female reproductive system. These side effects have been underestimated, although they are significant, requiring surgical or intensive medical treatment. Pre- and posttransplant gynecological evaluation should be performed to address the development of complications secondary to sirolimus in order to intervene sooner with alternative therapies.

Key words: Type 1 diabetes; Islet transplantation; Immunosuppression; Sirolimus; Reproductive system

Address correspondence to Dr. Rodolfo Alejandro, Diabetes Research Institute, University of Miami-Miller School of Medicine, 1450 NW 10th Avenue (R134), Miami, FL 33136, USA. Tel: (305) 243-6504; Fax: (305) 243-1058; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it