Cell Transplantation 24(2) Abstracts

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Cell Transplantation, Vol. 24, pp. 133-149, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675728
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Recent Advances in Mesenchymal Stem Cell Immunomodulation: The Role of Microvesicles

Alessandra Fierabracci,* Andrea Del Fattore,* Rosa Luciano,* Marta Muraca,† Anna Teti,‡ and Maurizio Muraca§

*Research Laboratories, Bambino Gesù Children’s Hospital, Rome, Italy
†School of Medicine, University “Sapienza”, Rome, Italy
‡Department of Biotechnological and Applied Clinical Sciences, University of L’Aquila, L’Aquila, Italy
§Department of Women's and Children's Health, University of Padua, Padua, Italy

Mesenchymal stem cells are the most widely used cell phenotype for therapeutic applications, the main reasons being their well-established abilities to promote regeneration of injured tissues and to modulate immune responses. Efficacy was reported in the treatment of several animal models of inflammatory and autoimmune diseases and, in clinical settings, for the management of disorders such as GVHD, systemic lupus erythematosus, multiple sclerosis, and inflammatory bowel disease. The effects of mesenchymal stem cells are believed to be largely mediated by paracrine signals, and several secreted molecules have been identified as contributors to the net biological effect. Recently, it has been recognized that bioactive molecules can be shuttled from cell to cell packed in microvesicles, tiny portions of cytoplasm surrounded by a membrane. Coding and noncoding RNAs are also carried in such microvesicles, transferring relevant biological activity to target cells. Several reports indicate that the regenerative effect of mesenchymal stem cells can be reproduced by microvesicles isolated from their culture medium. More recent evidence suggests that the immunomodulatory effects of mesenchymal stem cells are also at least partially mediated by secretedmicrovesicles. These findings allow better understanding of the mechanisms involved in cell-to-cell interaction and may have interesting implications for the development of novel therapeutic tools in place of the parent cells.

Key words: Microvesicles; Exosomes; Mesenchymal stem cells; Immunity

Received July 11, 2013; final acceptance November 20, 2013. Online prepub date: November 21, 2013.
Address correspondence to Alessandra Fierabracci, Ph.D., Research Laboratories, Children’s Hospital Bambino Gesù, Piazza Sant’Onofrio 4, 00165 Rome, Italy. Tel: +39 06 6859 2656; Fax: +39 06 6859 2904; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 151-165, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675142
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bone Marrow Mesenchymal Stromal Cells Isolated From Multiple Sclerosis Patients Have Distinct Gene Expression Profile and Decreased Suppressive Function Compared With Healthy Counterparts

Gislane L. V. de Oliveira,*† Kalil W. A. de Lima,*† Amanda M. Colombini,* Daniel G. Pinheiro,* Rodrigo A. Panepucci,* Patrícia V. B. Palma,* Doralina G. Brum,‡ Dimas T. Covas,*§ Belinda P.Simões,*§ Maria C. de Oliveira,*§ Eduardo A. Donadi,§ and Kelen C. R. Malmegrim*¶

*Center for Cell-Based Research, Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo (USP), Ribeirão Preto, São Paulo, Brazil
†Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo (USP), Ribeirão Preto, São Paulo, Brazil
‡Department of Neurology, Psychology and Psychiatry, School of Medicine of Botucatu, University of State of São Paulo (UNESP), Botucatu, São Paulo, Brazil
§Department of Clinical Medicine, Ribeirão Preto Medical School, University of São Paulo (USP), Ribeirão Preto, São Paulo, Brazil
¶Department of Clinical, Toxicological and Bromatological Analysis, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, São Paulo, Brazil

Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease of the central nervous system, due to an immune reaction against myelin proteins. Multipotent mesenchymal stromal cells (MSCs) present immunosuppressive effects and have been used for the treatment of autoimmune diseases. In our study, gene expression profile and in vitro immunomodulatory function tests were used to compare bone marrow-derived MSCs obtained from MS patients, at pre- and postautologous hematopoietic stem cell transplantation (AHSCT) with those from healthy donors. Patient MSCs comparatively exhibited i) senescence in culture; ii) similar osteogenic and adipogenic differentiation potential; iii) decreased expression of CD105, CD73, CD44, and HLA-A/B/C molecules; iv) distinct transcription at pre-AHSCT compared with control MSCs, yielding 618 differentially expressed genes, including the downregulation of TGFB1 and HGF genes and modulation of the FGF and HGF signaling pathways; v) reducedantiproliferative effects when pre-AHSCT MSCs were cocultured with allogeneic T-lymphocytes; vi) decreased secretion of IL-10 and TGF-β in supernatants of both cocultures (pre- and post-AHSCT MSCs); and vii) similar percentages of regulatory cells recovered after MSC cocultures. The transcriptional profile of patient MSCs isolated 6 months posttransplantation was closer to pre-AHSCT samples than from healthy MSCs. Considering that patient MSCs exhibited phenotypic changes, distinct transcriptional profile and functional defects implicated in MSC immunomodulatory and immunosuppressive activity, we suggest that further MS clinical studies should be conducted using allogeneic bone marrow MSCs derived from healthy donors. We also demonstrated that treatment of MS patients with AHSCT does not reverse the transcriptional and functional alterations observed in patient MSCs.

Key words: Multiple sclerosis (MS); Multipotent mesenchymal stromal cells (MSCs); Hematopoietic stem cell transplantation; Gene expression profile; Immunomodulatory and immunosuppressive activity

Received September 4, 2013; final acceptance November 5, 2013. Online prepub date: November 20, 2013.
Address correspondence to Gislane Lelis Vilela de Oliveira, Ph.D., Fundação Hemocentro de Ribeirão Preto, Rua Tenente Catão Roxo, 2501 Ribeirão Preto, São Paulo 14051-140, Brazil. Tel: +55-16-21019605; Fax: +55-16-21019309; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 167-182, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X674666
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Enhanced Recovery From Chronic Ischemic Injury by Bone Marrow Cells in a Rat Model of Ischemic Stroke

Jongman Yoo,1 Jin-Ju Seo,1 Jang-Hyeon Eom, and Dong-Youn Hwang

Department of Biomedical Science, College of Life Science, CHA University, Gyeonggido, Korea

Even after decades of intensive studies, therapeutic options for patients with stroke are rather limited. Thrombolytic drugs effectively treat the very acute stage of stroke, and several neuroprotectants that are designed to treat secondary injury following stroke are being tested in clinical trials. However, these pharmacological approaches primarily focus on acute stroke recovery, and few options are available for treating chronic stroke patients. In recent years, stem cell-mediated regenerative approaches have emerged as promising therapeutic strategies for treating the chronic stage of stroke. In this study, we examined whether systemically administered bone marrow cells (BMCs) could have beneficial effects in a rat model of chronic ischemia. Our transplantation experiments using BMCs obtained from ischemic donor rats showed functional and structural recovery during the chronic stage of stroke. BMC-mediated neural proliferation was prominent in the brains of rats with chronic stroke, and most of the new cells eventually became neurons instead of astrocytes. BMC-mediated enhanced neural proliferation coincided with a significant reduction (~50%) in the number of activated microglia, which is consistent with previous reports of enhanced neural proliferation being linked to microglial inactivation. Strikingly, approximately 57% of the BMCs that infiltrated the chronic ischemic brain were CD25+ cells, suggesting that these cells may exert the beneficial effects associated with BMC transplantation. Based on the reported anti-inflammatory role of CD25+ regulatory T-cells in acute experimental stroke, we propose a working model delineating the positive effects of BMC transplantation during the chronic phase of stroke; infiltrating BMCs (mostly CD25+ cells) reduce activated microglia, which leads to enhanced neural proliferation and enhanced recovery from neuronal damage in this rat model of chronic stroke. This study provides valuable insights into the effect of BMC transplantation in the chronic ischemic brain, which may lead to the development of effective therapy for chronic stroke patients who currently lack satisfactory therapeutic options.

Key words: Stroke; Bone marrow cells (BMCs); Ischemia; Neural proliferation; Inflammation

Received December 1, 2011; final acceptance October 11, 2013. Online prepub date: October 22, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Dong-Youn Hwang, Rm. 601, CHA Bio Complex, 335 Pankyo-ro, Bundang-gu, Gyeonggi-Do, Korea. Tel: +82-31-881-7142; Fax: +82-31-881-7242; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 183-190, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675755
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Long-Term Effects of Magnetically Targeted Ferumoxide-Labeled Human Neural Stem Cells in Focal Cerebral Ischemia

Miyeoun Song,*† Young-Ju Kim,* Yoon-ha Kim,* Jina Roh,* Eun-Cheol Kim,† Hong Jun Lee,‡ Seung U. Kim,‡§ and Byung-Woo Yoon*¶

*Department of Neurology, Clinical Research Institute, Seoul National University Hospital, Seoul, Korea
†Research Center for Tooth and Periodontal Regeneration, Kyung Hee University School of Dentistry, Seoul, Korea
‡Medical Research Institute, Chung-Ang University College of Medicine, Seoul, Korea
§Division of Neurology, Department of Medicine, University of British Columbia, Vancouver, BC, Canada
¶Neuroscience Research Institute, Seoul National University College of Medicine, Seoul, Korea

The long-term effect of magnetically targeted neural stem cells in a rat focal cerebral ischemia model was investigated. In middle cerebral artery occlusion (MCAO) stroke model rats, ferumoxide-labeled human neural stem cells (hNSCs) were injected into the tail vein. MCAO rats were divided into three groups: ischemia only (IO), ischemia with NSC injection (IC), and ischemia with NSC injection and the use of magnet targeting (IM). Four weeks after MCAO and 3 weeks posttransplantation, a greater number ofhNSCs were found in ischemic lesion sites in IM rat brain compared with IO and IC animals. In addition, differentiation of hNSCs into neurons or astrocytes and angiogenesis were markedly increased. In IM rats, infarct volume was considerably reduced, and function was significantly improved. The present study indicates that long-term use of magnetic fields may be a useful way to improve the efficacy of targeted migration of stem cells and functional deficits in stem cell-based therapy for ischemic brain injury.

Key words: Ferumoxide (iron oxide); Focal cerebral ischemia; Human neural stem cell (hNSCs); Magnet targeting; Cell therapy

Received September 10, 2013; final acceptance December 2, 2013. Online prepub date: December 30, 2013.
Address correspondence to Byung-Woo Yoon, M.D., Ph.D., Department of Neurology, Seoul National University Hospital, Seoul 110-744, Korea. Tel: +82-2-2072-2875; Fax: +82-2-3673-1990; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Seung U. Kim, M.D., Ph.D., Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, BC V6T 2B5, Canada. Tel: +1 604-822-7897; Fax: +1 604-822-7897; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 191-201, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X678526
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Long-Term Survival and Differentiation of Human Neural Stem Cells in Nonhuman Primate Brain With no Immunosuppression

Sang-Rae Lee,*1 Hong J. Lee,†1 Sang-Hoon Cha,‡1 Kang-Jin Jeong,* Youngjeon Lee,* Chang-Yeop Jeon,* Kyung Sik Yi,‡ Inja Lim,§ Zang-Hee Cho,¶ Kyu-Tae Chang,* and Seung U. Kim†#

*National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang, Korea
†Medical Research Institute, Chung-Ang University College of Medicine, Seoul, Korea
‡Department of Radiology, Chungbuk National University College of Medicine, Cheongju, Korea
§Departmentof Physiology, Chung-Ang University College of Medicine, Seoul, Korea
¶Neuroscience Research Institute, Gachon University of Medicine and Science, Inchon, Korea
#Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, BC, Canada

Cellular fate of human neural stem cells (hNSCs) transplanted in the brain of nonhuman primates (NHPs) with no immunosuppression was determined at 22 and 24 months posttransplantation (PTx) regarding survival, differentiation, and tumorigenesis. Survival of hNSCs labeled with magnetic nanoparticles was successfully detected around injection sites in the brain at 22 months PTx by MRI. Histological examination of brain sections with H&E and Prussian blue staining at 24 months revealed that most of the grafted hNSCs were found located along the injection tract. Grafted hNSCs were found to differentiate into neurons at 24 months PTx. In addition, none of the grafted hNSCs were bromodeoxyuridine positive in the monkey brain, indicating that hNSCsdid not replicate in the NHP brain and did not cause tumor formation. This study serves as a proof of principle and provides evidence that hNSCs transplanted in NHP brain could survive and differentiate into neurons in the absence of immunosuppression. It also serves as a preliminary study in our scheduled preclinical studies of hNSC transplantation in NHP stroke models.

Key words: Nonhuman primate (NHP); Human neural stem cells (hNSCs); Brain transplantation; Magnetic nanoparticle (MNP); Magnetic resonance imaging (MRI)

Received August 29, 2013; final acceptance December 30, 2013. Online prepub date: January 29, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Kyu-Tae Chang, Ph.D., National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang 363-883, Korea. Tel: +82-43-240-6300; Fax: +82-43-240-6309; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Seung U. Kim, M.D., Ph.D., Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, BC V6T2B5, Canada. Tel: +1 604-822-7145; Fax: +1 604-822-7897; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 203-211, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676934
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Regeneration Potential After Human and Autologous Stem Cell Transplantation in a Rat Sciatic Nerve Injury Model Can Be Monitored by MRI

Mathias Tremp,*†‡1 Moritz Meyer zu Schwabedissen,*†1 Elisabeth A. Kappos,*† Patricia E. Engels,*† Arne Fischmann,§ Arnaud Scherberich,¶ Dirk J. Schaefer,* and Daniel F. Kalbermatten*†

*Department of Plastic, Reconstructive, Aesthetic and Handsurgery, University of Basel Hospital, Basel, Switzerland
†Institute of Pathology, Department of Neuropathology, University of Basel Hospital, Basel, Switzerland
‡Department of Integrative Medical Biology, Section for Anatomy, Umeå University, Umeå, Sweden
§Department of Radiology, Division of Neuroradiology, University of Basel Hospital, Basel, Switzerland
¶Institute for Surgical Research and Hospital Management, University of Basel Hospital, Basel, Switzerland

Traumatic nerve injuries are a major clinical challenge. Tissue engineering using a combination of nerve conduits and cell-based therapies represents a promising approach to nerve repair. The aim of this study was to examine the regeneration potential of human adipose-derived stem cells (hASCs) after transplantation in a nonautogenous setting and to compare them with autogenous rat ASCs (rASCs) for early peripheral nerve regeneration. Furthermore, the use of MRI to assess the continuous process of nerve regeneration was elaborated. The sciatic nerve injury model in female Sprague–Dawley rats was applied, and a 10-mm gap created by using a fibrin conduit seeded with the following cell types: rASCs, Schwann cell (SC)-like cells from rASC, rat SCs (rSCs),hASCs from the superficial and deep abdominal layer, as well as human stromal vascular fraction (1 × 106 cells). As a negative control group, culture medium only was used. After 2 weeks, nerve regeneration was assessed by immunocytochemistry. Furthermore, MRI was performed after 2 and 4 weeks to monitor nerve regeneration. Autogenous ASCs and SC-like cells led to accelerated peripheral nerve regeneration, whereas the human stem cell groups displayed inferior results. Nevertheless, positive trends could be observed for hASCs from the deep abdominal layer. By using a clinical 3T MRI scanner, we were able to visualize the graft as a small black outline and small hyperintensity indicating the regenerating axon front. Furthermore, a strong correlation was found between the length of the regenerating axon front measured by MRI and the length measured by immunocytochemistry (r = 0.74, p = 0.09). We successfully transplanted and compared human and autologous stem cells for peripheral nerve regeneration in a rat sciatic nerve injury model. Furthermore, we were able to implement the clinical 3T MRI scanner to monitor the efficacy of cellular therapy over time.

Key words: Adult stem cells; Tissue engineering; Nerve regeneration; Transplantation; Heterologous

Received August 2, 2013; final acceptance December 20, 2013. Online prepub date: December 30, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Daniel F. Kalbermatten, M.D., Ph.D., Medical Director, Plastic, Reconstructive, Aesthetic and Handsurgery, University Basel, Spitalstrasse 21, 4031 Basel, Switzerland. Tel: +41 61 265 25 25; Fax: +41 61 265 73 01; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 213-222, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675700
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Stem Cell Salvage of Injured Peripheral Nerve

Nadia Grimoldi,* Federica Colleoni,† Francesca Tiberio,* Ignazio G. Vetrano,* Alberto Cappellari,‡ Antonella Costa,§ Marzia Belicchi,† Paola Razini,† Rosaria Giordano,¶ Diego Spagnoli,# MauroPluderi,* Stefano Gatti,** Michela Morbin,†† Sergio M. Gaini,* Paolo Rebulla,¶ Nereo Bresolin,† and Yvan Torrente†‡‡

*Unit of Neurosurgery, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Università degli Studi di Milano, Milan, Italy
†Stem Cell Laboratory, Department of Pathophysiology and Transplantation, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Università degli Studi di Milano, Centro Dino Ferrari, Milan, Italy
‡Unit of Neurophysiopathology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy
§Unit of Neuroradiology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy
¶Center for Transfusion Medicine, Cellular Therapy and Cryobiology, Department of Regenerative Medicine, Foundation IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy
#Unit of Neurosurgery, Moriggia Pelascini Hospital, Gravedona, Como, Italy
**Centre for Surgical Research, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Università degli Studi di Milano, Italy
††Unit of Neuropathology, Foundation IRCCS Neurological Institute “Carlo Besta,” Milan, Italy
‡‡UNISTEM, Centro Interdipartimentale di Ricerca sulle Cellule Staminali, Università degli Studi di Milano, Milan, Italy

We previously developed a collagen tube filled with autologous skin-derived stem cells (SDSCs) for bridging long rat sciatic nerve gaps. Here we present a case report describing a compassionate use of this graft for repairing the polyinjured motor and sensory nerves of the upper arms of a patient. Preclinical assessment was performed with collagen/SDSC implantation in rats after sectioning the sciatic nerve. For the patient, during the 3-year follow-up period, functional recovery of injured median and ulnar nerves was assessed by pinch gauge test and static two-point discrimination and touch test with monofilaments, along with electrophysiological and MRI examinations. Preclinical experiments in rats revealed rescue of sciatic nerve and no side effects of patient-derived SDSC transplantation (30 and 180 days of treatment). In the patient treatment, motor and sensory functions of the median nerve demonstrated ongoing recovery postimplantation during the follow-up period. The results indicate that the collagen/SDSC artificial nerve graft could be used for surgical repair of larger defects in major lesions of peripheral nerves, increasing patient quality of life by saving the upper arms from amputation.

Key words: Nerve graft; Collagen tube; Sciatic nerve; Skin-derived stem cells (SDSCs)

Received July 10, 2013; final acceptance November 14, 2013. Online prepub date: November 21, 2013.
Address correspondence to Dr. Yvan Torrente, Stem Cell Laboratory, Department of Pathophysiology and Transplantation, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Università degli Studi di Milano, Centro Dino Ferrari, Via F. Sforza 35, 20122 Milan, Italy. Tel: +39-02-55033874; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 223-233, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676196
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Distinct In Vitro Properties of Embryonic and Extraembryonic Fibroblast-Like Cells Are Reflected in Their In Vivo Behavior Following Grafting in the Adult Mouse Brain

Roberta Costa,* Irene Bergwerf,†‡ Eva Santermans,§ Nathalie De Vocht,†‡ Jelle Praet,†‡ Jasmijn Daans,†‡ Debbie Le Blon,†‡ Chloé Hoornaert,†‡ Kristien Reekmans,†‡ Niel Hens,§¶ Herman Goossens,‡Zwi Berneman,†‡ Ornella Parolini,# Francesco Alviano,* and Peter Ponsaerts†‡

*Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy
†Experimental Cell Transplantation Group, Laboratory of Experimental Hematology, University of Antwerp, Antwerp, Belgium
‡Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Antwerp, Belgium
§Center for Statistics, I-Biostat, Hasselt University, Hasselt, Belgium
¶Centre for Health Economic Research and Modeling Infectious Diseases (Chermid), University of Antwerp, Antwerp, Belgium
#Centro di Ricerca E. Menni, Fondazione Poliambulanza – Istituto Ospedaliero, Brescia, Italy

Although intracerebral transplantation of various fibroblast(-like) cell populations has been shown feasible, little is known about the actual in vivo remodeling of these cellular grafts and their environment. In this study, we aimed to compare the in vitro and in vivo behavior of two phenotypically similar—but developmentally distinct—fibroblast-like cell populations, namely, mouse embryonic fibroblasts (mEFs) and mouse fetal membrane-derived stromal cells (mFMSCs). While both mEFs and mFMSCs are readily able to reduce TNF-α secretion by LPS/IFN-γ-activated BV-2 microglia, mFMSCs and mEFs display strikingly opposite behavior with regard to VEGF production under normal and inflammatory conditions. Whereas mFMSCs downregulate VEGF production upon coculture with LPS/IFN-γ-activated BV-2 microglia, mEFs upregulate VEGF production in the presence of LPS/IFN-γ-activated BV-2 microglia. Subsequently, in vivo grafting of mFMSCs and mEFs revealed no difference in microglial andastroglial responses toward the cellular grafts. However, mFMSC grafts displayed a lower degree of neoangiogenesis compared to mEF grafts, thereby potentially explaining the lower cell number able to survive in mFMSC grafts. In summary, our results suggest that physiological differences between fibroblast-like cell populations might lie at the basis of variations in histopathological and/or clinical outcome following cell grafting in mouse brain.

Key words: Fetal membrane-derived stromal cells; Embryonic fibroblasts; Immunomodulation; Transplantation; Brain; Angiogenesis

Received September 3, 2013; final acceptance December 3, 2013. Online prepub date: December 30, 2013.
Address correspondence to Prof. Dr. Peter Ponsaerts, Experimental Cell Transplantation Group, Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Campus Drie Eiken (CDE-S6.51), Universiteitsplein 1, 2610 Antwerp (Wilrijk), Belgium. Tel: +32-3-2652428; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 235-245, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676204
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Engineering a Fibrocartilage Spectrum Through Modulation of Aggregate Redifferentiation

Meghan K. Murphy,* Taylor E. Masters,* Jerry C. Hu,* and Kyriacos A. Athanasiou*†

*Department of Biomedical Engineering, University of California Davis, Davis, CA, USA
†Department of Orthopaedic Surgery, University of California Davis, Davis, CA, USA

Expanded costochondral cells provide a clinically relevant cell source for engineering both fibrous and hyaline articular cartilage. Expanding chondrocytes in a monolayer results in a shift toward a proliferative, fibroblastic phenotype. Three-dimensional aggregate culture may, however, be used to recover chondrogenic matrix production. This study sought to engineer a spectrum of fibrous to hyaline neocartilage from a single cell source by varying the duration of three-dimensional culture following expansion. In third passage porcine costochondral cells, the effects of aggregate culture duration were assessed after 0, 8, 11, 14, and 21 days of aggregate culture and after 4 subsequent weeks of neocartilage formation. Varying the duration of aggregateredifferentiation generated a spectrum of fibrous to hyaline neocartilage. Within 8 days of aggregation, proliferation ceased, and collagen and glycosaminoglycan production increased, compared with monolayer cells. In self-assembled neocartilage, type II-to-I collagen ratio increased with increasing aggregate duration, yet glycosaminoglycan content varied minimally. Notably, 14 days of aggregate redifferentiation increased collagen content by 25%, tensile modulus by over 110%, and compressive moduli by over 50%, compared with tissue formed in the absence of redifferentiation. A spectrum of fibrous to hyaline cartilage was generated using a single, clinically relevant cell source, improving the translational potential of engineered cartilage.

Key words: Costal chondrocytes; Monolayer expansion; Three-dimensional culture; Self-Assembly; Hyaline cartilage

Received May 27, 2013; final acceptance December 4, 2013. Online prepub date: December 30, 2013.
Address correspondence to Kyriacos A. Athanasiou, University of California Davis, One Shields Ave., Davis, CA 95616, USA. Tel: +1 530 754 6645; Fax: +1 530 754 5739; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 247-261, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675160
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Therapeutic Potential of Laminin–Biodritin Microcapsules for Type 1 Diabetes Mellitus

Ana Lucia Campanha-Rodrigues,* Gisella Grazioli,*† Talita C. Oliveira,* Ana Carolina V. Campos-Lisbôa,*† Thiago R. Mares-Guia,*† and Mari C. Sogayar*

*Chemistry Institute, Biochemistry Department, Cell and Molecular Therapy Center (NUCEL/NETCEM), School of Medicine, University of São Paulo, São Paulo, SP, Brazil
†Cell Protect Biotechnology Ltda., São Paulo, SP, Brazil

Pancreatic islet microencapsulation constitutes an attractive therapy for type 1 diabetes mellitus; however, long-term β-cell function remains a major problem. Loss of extracellular matrix interactions during islet isolation dramatically affects β-cell viability. We have previously shown beneficial effects of laminin (LN) in human islet cultures. Herein, we investigated whether LN could improve the outcome of transplantation after islet microencapsulation in Biodritin, an alginate-based material. To test LN-Biodritinstability, microcapsules were subjected to different types of in vitro stress. Focusing on biocompatibility, empty microcapsules were coincubated with the RAW 264.7 macrophage cell line for up to 24 h, and empty beads were implanted IP in mice and retrieved for analyses after 7 and 30 days. Upon culturing for 48 h, mRNA, protein levels, and caspase 3 activity were evaluated in islets microencapsulated with LN-Biodritin. Mice rendered diabetic by streptozotocin injection were transplanted with microencapsulated islets, followed by assessment of body weight, glycemia, and graft function (evaluated by OGTT). Graft efficiency was observed upon microencapsulated islet explantation. The results obtained showed that LN-Biodritin microcapsules were as stable and biocompatible as Biodritin. Modulation of mRNA and protein levels suggested protection against apoptosis and islet stress. Mice transplanted with LN-Biodritin microencapsulated islets presented a better outcome at 198 days postsurgery. Graftexplantation led animals to hyperglycemia. In conclusion, LN-Biodritin constitutes a very promising biomaterial for islet transplantation.

Key words: Laminin (LN); Alginate; Type 1 diabetes mellitus; Islet transplantation; Microencapsulation; Biomaterials

Received November 9, 2012; final acceptance November 6, 2013. Online prepub date: November 20, 2013.
Address correspondence to Professor Mari C. Sogayar, Ph.D., Chemistry Institute, University of São Paulo, Av. Professor Lineu Prestes, 748, Bloco 9S, São Paulo, 05508-000 SP, Brazil. Tel: +55 (11) 2648-0226; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 263-276, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X678535
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Immunological Shielding by Induced Recruitment of Regulatory T-Lymphocytes Delays Rejection of Islets Transplanted in Muscle

Evelina Vågesjö,* Gustaf Christoffersson,* Tomas B. Waldén,* Per-Ola Carlsson,* Magnus Essand,† Olle Korsgren,† and Mia Phillipson*

*Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden
†Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden

The only clinically available curative treatment of type 1 diabetes mellitus is replacement of the pancreatic islets by allogeneic transplantation, which requires immunosuppressive therapies. Regimens used today are associated with serious adverse effects and impaired islet engraftment and function. The aim of the current study was to induce local immune privilege by accumulating immune-suppressive regulatory T-lymphocytes (Tregs) at the site of intramuscular islet transplantation to reduce the need of immunosuppressive therapy during engraftment. Islets were cotransplanted with a plasmid encoding the chemokine CCL22 into the muscle of MHC-mismatched mice, after which pCCL22 expression and leukocyte recruitment were studied in parallel with graft functionality. Myocyte pCCL22 expression and secretion resulted in local accumulation of Tregs. When islets were cotransplanted with pCCL22, significantly fewer effector T-lymphocytes were observed in close proximity to the islets, leading to delayed graft rejection. As a result, diabetic recipients cotransplanted with islets and pCCL22 intramuscularly became normoglycemic for 10 consecutive days, while grafts cotransplanted with control plasmid were rejected immediately, leaving recipients severely hyperglycemic. Here we propose a simple method to initially shield MHC-mismatched islets by the recruitment of endogenous Tregs during engraftment in order to improve early islet survival. Using this approach, the very high doses of systemic immunosuppression used initially following transplantation can thereby be avoided.


Key words: Diabetes; Immunosuppression; C-C motif ligand 22 (CCL22); DNA plasmid; Muscle; Intramuscular transplantation

Received May 7, 2013; final acceptance January 8, 2014. Online prepub date: January 29, 2014.
Address correspondence to Mia Phillipson, Department of Medical Cell Biology, Uppsala University, P.O. Box 571, 751 23 Uppsala, Sweden. Tel: +46 184714419; Fax: +46 184714938; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 277-286, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X674053
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Restoration of Anal Sphincter Function After Myoblast Cell Therapy in Incontinent Rats

Aurélie Bisson,*†‡ Manuel Fréret,*† Laurent Drouot,*† Laetitia Jean,*† Stéphanie Le Corre,*§ Guillaume Gourcerol,†¶ Christelle Doucet,‡ Francis Michot,†¶# Olivier Boyer,*†§1 and Marek Lamacz*†1

*Inserm, U905, Rouen, France
†Normandy University, IRIB, Rouen, France
‡Celogos, Paris, France
§Rouen University Hospital, Laboratory of Biotherapy, Rouen, France
¶Inserm, U1073, Rouen, France
#Rouen University Hospital, Department of Digestive Surgery, Rouen, France

Fecal incontinence (FI) remains a socially isolating condition with profound impact on quality of life for which autologous myoblast cell therapy represents an attractive treatment option. We developed an animal model of FI and investigated the possibility of improving sphincter function by intrasphincteric injection of syngeneic myoblasts. Several types of anal cryoinjuries were evaluated on anesthetized Fischer rats receiving analgesics. The minimal lesion yielding sustainable anal sphincter deficiency was a 90° cryoinjury of the sphincter, repeated after a 24-h interval. Anal sphincter pressure was evaluated longitudinally by anorectal manometry under local electrostimulation. Myoblasts were prepared using a protocol mimicking a clinical-grade process and further transduced with a GFP-encoding lentiviral vector before intrasphincteric injection. Experimental groups were uninjured controls, cryoinjured + PBS, and cryoinjured + myoblasts (different doses or injection site). Myoblast injection was well tolerated. Transferred myoblasts expressing GFP integrated into the sphincter and differentiated in situ into dystrophin-positive mature myofibers. Posttreatment sphincter pressures increased over time. At day 60, pressures in the treated group were significantly higher than those of PBS-injected controls and not significantly different from those of normal rats. Longitudinal follow-up showed stability of the therapeutic effect on sphincter function over a period of 6 months. Intrasphincteric myoblast injections at the lesion borders were equally as effective as intralesion administration, but an injection opposite to the lesion was not. These results provide proof of principle for myoblast cell therapy to treat FI in a rat model. This strategy is currently being evaluated in humans in a randomized double-blind placebo-controlled clinical trial.

Key words: Fecal incontinence (FI); Anal sphincter; Myoblasts; Cell therapy; Experimental model

Received December 31, 2012; final acceptance September 16, 2013. Online prepub date: October 18, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Olivier Boyer, Inserm U905, Faculty of Medicine and Pharmacy, 22 bd Gambetta, F-76000 Rouen, France. Tel: +33 235 148535; Fax: +33 235 148494; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 287-304, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X675719
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Propagation of Human Corneal Endothelial Cells: A Novel Dual Media Approach

Gary S. L. Peh,* Zhenzhi Chng,† Heng-Pei Ang,* Terence Y. D. Cheng,† Khadijah Adnan,* Xin-Yi Seah,* Benjamin L. George,* Kah-Peng Toh,* Donald T. Tan,*‡§ Gary H. F. Yam,* Alan Colman,† andJodhbir S. Mehta*‡¶

*Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore, Singapore
†A*STAR Institute of Medical Biology, Singapore, Singapore
‡Singapore National Eye Centre, Singapore, Singapore
§Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore
¶Duke Medical School of Medicine, National University of Singapore, Singapore, Singapore

Corneal endothelium-associated corneal blindness is the most common indication for corneal transplantation. Restorative corneal transplant surgery is the only option to reverse the blindness, but a global shortage of donor material remains an issue. There are immense clinical interests in the development of alternative treatment strategies to alleviate current reliance on donor materials. For such endeavors, ex vivo propagation of human corneal endothelial cells (hCECs) is required, but current methodology lacks consistency, with expanded hCECs losing cellular morphology to a mesenchymal-like transformation. In this study, we describe a novel dual media culture approach for the in vitro expansion of primary hCECs. Initial characterization included analysis of growth dynamics of hCECs grown in either proliferative (M4) or maintenance (M5) medium. Subsequent comparisons were performed on isolated hCECs cultured in M4 alone against cells expanded using the dual media approach. Further characterizations were performed using immunocytochemistry, quantitative real-time PCR, and gene expression microarray. At the third passage, results showed that hCECs propagated using the dual media approach were homogeneous in appearance, retained their unique polygonal cellular morphology, and expressed higher levels of corneal endothelium-associated markers in comparison to hCECs cultured in M4 alone, which were heterogeneous and fibroblastic in appearance. Finally, for hCECs cultured using the dual media approach, global gene expression and pathway analysis between confluent hCECs before and after 7-day exposure to M5 exhibited differential gene expression associated predominately with cell proliferation and wound healing. These findings showed that the propagation of primary hCECs using the novel dual media approach presented in this study is a consistent method to obtain bona fide hCECs. This, in turn, will elicit greater confidence in facilitating downstream development of alternative corneal endothelium replacement using tissue-engineered graft materials or cell injection therapy.

Key words: Cornea; Human corneal endothelial cells (hCECs); Cell culture; Cell transplantation; Tissue engineering

Received September 3, 2013; final acceptance November 20, 2013. Online prepub date: November 21, 2013.
Address correspondence to Adj. Assoc. Professor Jodhbir Mehta, Singapore National Eye Centre, 11 Third Hospital Avenue, #08-00, Singapore 168751, Singapore. Tel: +65 6322 4571; Fax: +65 6323 1903; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 287-304, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X678544
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Attached Segment Has Higher CD34+ Cells and CFU-GM Than the Main Bag After Thawing

Hye Ryun Lee,*1 Sue Shin,†‡§1 Jong Hyun Yoon,†‡§ Eun Youn Roh,†‡§ Eun Young Song,† Kyou Sup Han,† and Byoung Jae Kim¶

*Department of Laboratory Medicine, Gyeongsang National University Hospital, Jinju, Korea
†Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea
‡Department of Laboratory Medicine, Seoul National University Boramae Hospital, Seoul, Korea
§Seoul Metropolitan Government Public Cord Blood Bank (Allcord), Seoul, Korea
¶Department of Obstetrics and Gynecology, Seoul National University Boramae Hospital, Seoul, Korea

A contiguous segment attached to the cord blood unit (CBU) is required for verifying HLA types, cell viability, and, possibly, potency before transplantation since such a segment is considered to be representative of the CBU. However, little is known regarding the characteristics of contiguous segments in comparison to main bag units due to the difficulty experienced in accessing a large number of cryopreserved CBUs. In this study, we used 245 nonconforming CBUs for allogeneic transplantation. After thawing the cryopreserved CBU, the number of total nucleated cells (TNCs), CD34+ cells, and CFUs in CB from main bags and segments, as well as cell viability and apoptosis, were examined. The comparative analysis showed that the number of TNCs was significantly higher in CB from main bags, whereas the numbers of CD34+ cells and CFU-GM were significantly higher in CB from segments. While the cell viability of TNCs in segments was higher, the proportion of apoptotic TNCs was also higher. In contrast, no difference was observed between the proportion of apoptotic CD34+ cells in main bags and segments. In the correlation analysis, the numbers of TNCs, CD34+ cells, and CFU-GM in main bags were highly correlated with those in segments, indicating that CB from segments is indeed representative of CB in main bags. Taken together, we conclude that segments have higher CD34+ cells and CFU-GM and lower TNCs than the main cryopreserved bag, although the two compartments are highly correlated with each other.

Key words: Cord blood; Cryopreservation; Quality

Received May 7, 2013; final acceptance January 9, 2014. Online prepub date: January 29, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Jong Hyun Yoon, M.D., Professor, Department of Laboratory Medicine, Seoul National University Boramae Hospital, Boramaro 5 gil 20, Dongjak-gu, 156-707 Seoul, Korea. Tel: +82-1-870-2601; Fax: +82-1-870-2630; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it