Cell Transplantation 23(11) Abstracts

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Cell Transplantation, Vol. 23, pp. 1321-1348, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670949
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Current Status of Islet Encapsulation

Lourdes Robles,* Rick Storrs,† Morgan Lamb,* Michael Alexander,* and Jonathan R. T. Lakey*‡

*Department of Surgery, University of California Irvine, Irvine, CA, USA
†Islet Sheet Medical, San Francisco, CA, USA
‡Biomedical Engineering, University of California Irvine, Irvine, CA, USA

Cell encapsulation is a method of encasing cells in a semipermeable matrix that provides a permeable gradient for the passage of oxygen and nutrients, but effectively blocks immune-regulating cells from reaching the graft, preventing rejection. This concept has been described as early as the 1930s, but it has exhibited substantial achievements over the last decade. Several advances in encapsulation engineering, chemical purification, applications, and cell viability promise to make this a revolutionary technology. Several obstacles still need to be overcome before this process becomes a reality, including developing a reliable source of islets or insulin-producing cells, determining the ideal biomaterial to promote graft function, reducing the host response to the encapsulation device, and ultimately a streamlined, scaled-up process for industry to be able to efficiently and safely produce encapsulated cells for clinical use. This article provides a comprehensive review of cell encapsulation of islets for the treatment of type 1 diabetes, including a historical perspective, current research findings, and future studies.

Key words: Cell encapsulation; Islets; Stem cells; Xenotransplantation; Cell engineering

Received November 2, 2012; final acceptance July 16, 2013. Online prepub date: July 22, 2013.
Address correspondence to Jonathan Lakey, Ph.D., MSM, Associate Professor of Surgery and Biomedical Engineering, Director of Research and Clinical Islet Program, Department of Surgery, University of California Irvine, 333 City Blvd. West, Suite 700, Orange, CA 92868, USA. Tel: +1-714-456-5386; Fax: +1-714-456-6377; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1349-1364, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X663550
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Improvement of Subcutaneous Bioartificial Pancreas Vascularization and Function by Coencapsulation of Pig Islets and Mesenchymal Stem Cells in Primates

Sophie Veriter,* Pierre Gianello,* Yasuhiro Igarashi,* Gwen Beaurin,* Audrey Ghyselinck,* Najima Aouassar,† Benedicte Jordan,† Bernard Gallez,† and Denis Dufrane*‡

*Universite Catholique de Louvain, Secteur des Sciences de la Sante, Institute of Clinical and Experimental Research, Pole de Chirurgie Experimentale et Transplantation, Brussels, Belgium
†University Clinical Hospital St Luc, Endocrine Cell Therapy Unit, Center of Tissular/Cellular Therapy, Brussels, Belgium
‡Universite Catholique de Louvain, Secteur des Sciences de la Sante, Louvain Drug Research Institute, Biomedical Magnetic Resonance Research Group, Brussels, Belgium

Insufficient oxygenation can limit the long-term survival of encapsulated islets in subcutaneous tissue. Transplantation of coencapsulated pig islets with adipose or bone marrow mesenchymal stem cells (AMSCs or BM-MSCs, respectively) was investigated with regard to implant vascularization, oxygenation, and diabetes correction in primates. The in vivo impact of MSCs on graft oxygenation and neovascularization was assessed in rats with streptozotocin (STZ)-induced diabetes that were subcutaneously transplanted with islets coencapsulated with AMSCs (n = 8) or BM-MSCs (n = 6). Results were compared to islets encapsulated alone (n = 8). STZ diabetic primates were subcutaneously transplanted with islets coencapsulated with BM-MSCs (n = 4) or AMSCs (n = 6). Recipients were monitored metabolically and immunologically, and neoangiogenesis was assessed on explanted grafts. Results were compared with primates transplanted with islets encapsulated alone (n = 5). The cotransplantation of islets with BM-MSCs or AMSCs in diabetic rats showed significantly higher graft oxygenation than islets alone (3% and 3.6% O2 for islets + BM-MSCs or AMSCs, respectively, vs. 2.2% for islets alone). A significantly better glycated hemoglobin correction (28 weeks posttransplantation) was found for primates transplanted with islets and MSCs (7.4% and 8.1%, respectively) in comparison to islets encapsulated alone (10.9%). Greater neoangiogenesis was found in the periphery of coencapsulated islets and AMSCs in comparison to islets alone (p < 0.01). In conclusion, the coencapsulation of pig islets with MSCs can improve significantly the islets’ survival/function in vitro. The coencapsulation of islets with MSCs improves implant oxygenation and neoangiogenesis. However, the cotransplantation of islets with MSCs improves only slightly the long-term function of a subcutaneous bioartificial pancreas in a primate preclinical model.

Key words: Bioartificial pancreas; Mesenchymal stem cells (MSCs); Oxygenation; Islets

Received April 26, 2012; final acceptance January 11, 2013. Online prepub date: February 4, 2013.
Address correspondence to Denis Dufrane, M.D., Ph.D., Universite Catholique de Louvain, Institute of Experimental and Clinical Research, Pole de Chirurgie Experimentale et Transplantation, Avenue Hippocrate 55, bte B1.55.04, B-1200 Brussels, Belgium. Tel: +32 (0)2 764 55 87; Fax: +32 (0)2 764 95 20; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1365-1379, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X672055
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Nonenzymatic Cryogenic Isolation of Therapeutic Cells: Novel Approach for Enzyme-Free Isolation of Pancreatic Islets Using In Situ Cryopreservation of Islets and Concurrent Selective Freeze Destruction of Acinar Tissue

Michael J. Taylor*† and Simona C. Baicu*‡

*Cell and Tissue Systems, Inc., N. Charleston, SC, USA
†Carnegie Mellon University, Pittsburgh, PA, USA
‡Current affiliation: Director of Ocular and Tissue Recovery Operations, LifePoint, Inc., Charleston, SC, USA

Cell-based therapies, which all involve processes for procurement and reimplantation of living cells, currently rely upon expensive, inconsistent, and even toxic enzyme digestion processes. A prime example is the preparation of isolated pancreatic islets for the treatment of type 1 diabetes by transplantation. To avoid the inherent pitfalls of these enzymatic methods, we have conceptualized an alternative approach based on the hypothesis that cryobiological techniques can be used for differential freeze destruction of the pancreas (Px) to release islets that are selectively cryopreserved in situ. Pancreata were procured from juvenile pigs using approved procedures. The concept of cryoisolation is based on differential processing of the pancreas in five stages: 1) infiltrating islets in situ preferentially with a cryoprotectant (CPA) cocktail via antegrade perfusion of the major arteries; 2) retrograde ductal infusion of water to distend the acinar; 3) freezing the entire Px solid to < −160°C for storage in liquid nitrogen; 4) mechanically crushing and pulverizing the frozen Px into small fragments; 5) thawing the frozen fragments, filtering, and washing to remove the CPA. Finally, the filtered effluent (cryoisolate) was stained with dithizone for identification of intact islets and with Syto 13/PI for fluorescence viability testing and glucose-stimulated insulin release assessment. As predicted, the cryoisolate contained small fragments of residual tissue comprising an amorphous mass of acinar tissue with largely intact and viable (>90%) embedded islets. Islets were typically larger (range 50–500 μm diameter) than their counterparts isolated from juvenile pigs using conventional enzyme digestion techniques. Functionally, the islets from replicate cryoisolates responded to a glucose challenge with a mean stimulation index = 3.3 ± 0.7. An enzyme-free method of islet isolation relying on in situ cryopreservation of islets with simultaneous freeze destruction of acinar tissue is feasible and proposed as a new and novel method that avoids the problems associated with conventional collagenase digestion methods.

Key words: Cryoisolation; Nonenzymatic preparation; Islets; Cryopreservation; Enzyme-free isolation

Received April 17, 2013; final acceptance August 26, 2013. Online prepub date: August 29, 2013.
Address correspondence to Michael J. Taylor, Ph.D., VP Research and Development, Cell and Tissue Systems, Inc., 2231 Technical Parkway, N. Charleston, SC 29406, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1381-1394, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670930
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Role of Alloresponsive Ly49+ NK Cells in Rat Islet Allograft Failure in the Presence and Absence of Cytomegalovirus

Maaike J. Smelt,* Marijke M. Faas,* Bart J. de Haan,* Aalzen de Haan,† John T. Vaage,‡ and Paul de Vos*

*Department of Pathology and Medical Biology, Division of Medical Biology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands
†Department of Medical Microbiology, Molecular Virology Section, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands
‡Institute of Immunology, Rikshospitalet University Hospital, Oslo, Norway

There are still many factors to discover to explain the low success rates of islet allografts. In this study, we demonstrate that specific subpopulations of alloreactive NK cells may be involved in the failure of islet allografts. By performing allotransplantation in rats (n = 13), we observed peripheral expansion and infiltration of alloreactive Ly49i2+ NK cells in the grafts. An effective strategy in rats to enhance the expansion of Ly49i2+ NK cells is performing a rat cytomegalovirus infection (n = 6). Cytomegalovirus infection was associated with an early expansion of the Ly49i2+ NK cells and accelerated islet graft failure. The Ly49i2+ NK cells are both alloreactive and involved in virus clearance. The expansion of this subpopulation could not be blocked by cyclosporin A immunosuppression. Also alloreactive KLRH1+ NK cells infiltrated the grafts, but nonalloreactive NKR-P1B+ cells were not observed in the islet allografts. Perforin staining of the infiltrating NK cells demonstrated the cytotoxic capacity of these cells. Our data suggest a role for this NK subpopulation in rat islet allograft destruction.

Key words: Islets; Natural killer (NK) cells; Allotransplantation; Immunosuppression; Cytomegalovirus

Received February 28, 2013; final acceptance July 10, 2013. Online prepub date: July 17, 2013.
Address correspondence to Paul de Vos, Ph.D., Professor, Pathology and Medical Biology, Section Immunoendocrinology, University Medical Center Groningen, University of Groningen, Hanzeplein 1, 9700 RB Groningen, the Netherlands. Tel: (+31) (50) 361 1045; Fax: (+31) (50) 3361 9911; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1395-1406, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670921
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Rat Mesenchymal Stem Cell Secretome Promotes Elastogenesis and Facilitates Recovery From Simulated Childbirth Injury

Charuspong Dissaranan,*† Michelle A. Cruz,† Matthew J. Kiedrowski,‡ Brian M. Balog,† Bradley C. Gill,*†§ Marc S. Penn,‡¶ Howard B. Goldman,*§ and Margot S. Damaser*†§#

*Glickman Urologic and Kidney Institute, Cleveland Clinic, Cleveland, OH, USA
†Department of Biomedical Engineering, Cleveland Clinic, Cleveland, OH, USA
‡Department of Integrative Medical Sciences, Northeast Ohio University College of Medicine, Rootstown, OH, USA
§Cleveland Clinic Lerner College of Medicine at Case Western Reserve University, Cleveland Clinic, Cleveland, OH, USA
¶Summa Cardiovascular Institute, Summa Health System, Akron, OH, USA
#Advanced Platform Technology Center, Louis Stokes Cleveland VA Medical Center, Cleveland, OH, USA

Vaginal delivery is a risk factor for stress urinary incontinence (SUI). Mesenchymal stem cells (MSCs) home to injured organs and can facilitate repair. The goal of this study was to determine if MSCs home to pelvic organs after simulated childbirth injury and facilitate recovery from SUI via paracrine factors. Three experiments were performed. Eighteen female rats received vaginal distension (VD) or sham VD and labeled intravenous (IV) MSCs to investigate if MSCs home to the pelvic organs. Whole-organ imaging and immunofluorescence were performed 1 week later. Thirty-four female rats received VD and IV MSCs, VD and IV saline, or sham VD and IV saline to investigate if MSCs accelerate recovery of continence. Twenty-nine female rats received VD and periurethral concentrated conditioned media (CCM), VD and periurethral control media, or sham VD and periurethral control media to investigate if factors secreted by MSCs accelerate recovery from VD. Urethral histology and function were assessed 1 week later. Significantly more MSCs were observed in the urethra, vagina, and spleen after VD compared to sham VD. Continence as measured by leak point pressure (LPP) was significantly reduced after VD in rats treated with saline or control media compared to sham VD but not in those given MSCs or CCM. External urethral sphincter (EUS) function as measured by electromyography (EMG) was not improved with MSCs or CCM. Rats treated with MSCs or CCM demonstrated an increase in elastin fibers near the EUS and urethral smooth muscle more similar to that of sham-injured animals than rats treated with saline or control media. MSCs homed to the urethra and vagina and facilitated recovery of continence most likely via secretion of paracrine factors. Both MSCs and CCM have promise as novel noninvasive therapies for SUI.

Key words: Paracrine factors; Urethra; External urethral sphincter (EUS); Electromyography (EMG); Female; Stress urinary incontinence (SUI); Elastin

Received July 2, 2012; final acceptance July 10, 2013. Online prepub date: July 17, 2013.
Address correspondence to Margot S. Damaser, Ph.D., Department of Biomedical Engineering, Cleveland Clinic, 9500 Euclid Ave. ND20, Cleveland, OH 44195, USA. Tel: +1-216-444-1103; Fax: +1-216-444-9198; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1407-1423, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670958
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Survival and Differentiation of Adenovirus-Generated Induced Pluripotent Stem Cells Transplanted Into the Rat Striatum

Kyle D. Fink,*†‡§¶#** Julien Rossignol,*††1 Ming Lu,*‡‡1 Xavier Lévêque,*‡¶#**‡‡ Travis D. Hulse,* Andrew T. Crane,* Veronique Nerriere-Daguin,‡¶#** Robert D. Wyse,* Phillip A. Starski,* Matthew T. Schloop,* Dylan J. Dues,* Steve J. Witte,* Cheng Song,* Ludovic Vallier,§§ Tuan H. Nguyen,‡#**¶¶ Philippe Naveilhan,‡¶#**## Ignacio Anegon,‡¶#** Laurent Lescaudron,†§¶#**,*** and Gary L. Dunbar*††‡‡

*Program in Neuroscience, Field Neurosciences Laboratory for Restorative Neurology Brain Research and Integrative Neuroscience Center, Central Michigan University, Mount Pleasant, MI, USA
†Faculté des Science et des Techniques, Université de Nantes, Nantes, France
‡INSERM U1064, ITUN, Nantes, France
§INSERM U791, Laboratoire d’Ingenierie Osteo-Articulaire et Dentaire (LIOAD), Nantes, France
¶INSERM UMR 643, Nantes, France
#Faculté de Médecine, Université de Nantes, Nantes, France
**Centre Hospitalier-Universitaire (CHU) HOTEL DIEU de Nantes, Nantes, France
††College of Medicine, Central Michigan University, Mount Pleasant, MI, USA
‡‡Field Neurosciences Institute, Saginaw, MI, USA
§§Wellcome Trust–Medical Research Council Stem Cell Institute, Anne McLaren Laboratory for Regenerative Medicine, Department of Surgery, University of Cambridge, Cambridge, UK/Wellcome Trust Sanger Institute, Hinxton, UK
¶¶INSERM U948, Biothérapies Hépatiques, Nantes, France
##INSERM UMR U913, Institut des Maladies de l’Appareil Digestif (IMAD), Nantes, France
***Faculté de Chirurgie Dentaire, Université de Nantes, Nantes, France

Induced pluripotent stem cells (iPSCs) offer certain advantages over embryonic stem cells in cell replacement therapy for a variety of neurological disorders. However, reliable procedures, whereby transplanted iPSCs can survive and differentiate into functional neurons, without forming tumors, have yet to be devised. Currently, retroviral or lentiviral reprogramming methods are often used to reprogram somatic cells. Although the use of these viruses has proven to be effective, formation of tumors often results following in vivo transplantation, possibly due to the integration of the reprogramming genes. The goal of the current study was to develop a new approach, using an adenovirus for reprogramming cells, characterize the iPSCs in vitro, and test their safety, survivability, and ability to differentiate into region-appropriate neurons following transplantation into the rat brain. To this end, iPSCs were derived from bone marrow-derived mesenchymal stem cells and tail-tip fibroblasts using a single cassette lentivirus or a combination of adenoviruses. The reprogramming efficiency and levels of pluripotency were compared using immunocytochemistry, flow cytometry, and real-time polymerase chain reaction. Our data indicate that adenovirus-generated iPSCs from tail-tip fibroblasts are as efficient as the method we used for lentiviral reprogramming. All generated iPSCs were also capable of differentiating into neuronal-like cells in vitro. To test the in vivo survivability and the ability to differentiate into region-specific neurons in the absence of tumor formation, 400,000 of the iPSCs derived from tail-tip fibroblasts that were transfected with the adenovirus pair were transplanted into the striatum of adult, immune-competent rats. We observed that these iPSCs produced region-specific neuronal phenotypes, in the absence of tumor formation, at 90 days posttransplantation. These results suggest that adenovirus-generated iPSCs may provide a safe and viable means for neuronal replacement therapies.

Key words: Induced pluripotent stem cells (iPSCs); Transplantation; Adenovirus; Neuronal differentiation; Stem cell

Received May 21, 2013; final acceptance July 16, 2013. Online prepub date: July 22, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Gary L. Dunbar, Field Neurosciences Laboratory for Restorative Neurology Brain Research and Integrative Neuroscience Center, Central Michigan University, Mount Pleasant, MI 48859, USA. Tel: +1-989-774-3282; Fax: +1-989-774-7108; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1425-1450, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670174
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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3-MA Inhibits Autophagy and Favors Long-Term Integration of Grafted Gad67–GFP GABAergic Precursors in the Developing Neocortex by Preventing Apoptosis

Christian Roux,* Céline Lesueur,*† Caroline Aligny,* Carole Brasse-Lagnel,*† Damien Genty,‡ Stéphane Marret,*§ Annie Laquerrière,*‡ Soumeya Bekri,*† and Bruno J. Gonzalez*

*NeoVasc Laboratory, ERI28, Microvascular Endothelium and Neonate Brain Lesions, Institute of Research for Innovation in Biomedicine, Normandy University, Rouen, France
†Department of Medical Biochemistry, Rouen University Hospital, Rouen, France
‡Department of Pathology, Rouen University Hospital, Rouen, France
§Department of Neonatal Paediatrics and Intensive Care, Rouen Hospital, Rouen, France

In human neonates, immature GABAergic interneurons are markedly affected by an excitotoxic insult. While in adults the interest of cell transplantation has been demonstrated in several neurological disorders, few data are available regarding the immature brain. The low survival rate constitutes a strong limitation in the capacity of transplanted neurons to integrate the host tissue. Because i) autophagy is an adaptive process to energetic/nutrient deprivation essential for cell survival and ii) literature describes cross-talks between autophagy and apoptosis, we hypothesized that regulation of autophagy would represent an original strategy to favor long-term survival of GABAergic precursors grafted in the immature neocortex. Morphological, neurochemical, and functional data showed that in control conditions, few grafted Gad67–GFP precursors survived. The first hours following transplantation were a critical period with intense apoptosis. Experiments performed on E15.5 ganglionic eminences revealed that Gad67–GFP precursors were highly sensitive to autophagy. Rapamycin and 3-MA impacted on LC3 cleavage, LC3II translocation, and autophagosome formation. Quantification of Bax, mitochondrial integrity, caspase-3 cleavage, and caspase-3 immunolocalization and activity showed that 3-MA induced a significant decrease of Gad67–GFP precursor apoptosis. In vivo, 3-MA induced, within the first 24 h, a diffuse LC3 pattern of grafted Gad67–GFP precursors, an increase of precursors with neurites, a reduction of the density of caspase-3 immunoreactive cells. A twofold increase in the survival rate occurred 15 days after the graft. Surviving neurons were localized in the cortical layers II–IV, which were still immature when the transplantation was done. Altogether, these data indicate that inhibition of autophagy represents an original strategy to allow GABAergic interneurons to overpass the first critical hours following transplantation and to increase their long-term survival in mice neonates.

Key words: Neonates; GABA interneurons; Autophagy; Bax; Caspases

Received September 28, 2012; final acceptance July 2, 2013. Online prepub date: July 11, 2013.
Address correspondence to Bruno J. Gonzalez, ERI28 NeoVasc, Laboratory of Microvascular Endothelium and Neonate Brain Lesions, Institute of Research for Innovation in Biomedicine, Normandy University, Rouen, France. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1451-1464, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670967
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Transplantation of Neural Stem/Progenitor Cells at Different Locations in Mice With Spinal Cord Injury

Hiroki Iwai,*† Satoshi Nori,‡ Soraya Nishimura,*† Akimasa Yasuda,§ Morito Takano,* Osahiko Tsuji,¶ Kanehiro Fujiyoshi,§ Yoshiaki Toyama,* Hideyuki Okano,† and Masaya Nakamura*

*Department of Orthopaedic Surgery, Keio University School of Medicine, Shinjuku, Tokyo, Japan
†Department of Physiology, Keio University School of Medicine, Shinjuku, Tokyo, Japan
‡Department of Orthopaedic Surgery, Tokyo Dental College Ichikawa General Hospital, Ichikawa, Chiba, Japan
§Department of Orthopaedic Surgery, National Hospital Organization, Murayama Medical Center, Musashimurayama, Tokyo, Japan
¶Department of Orthopaedic Surgery, Saitama Social Insurance Hospital, Urawa, Saitama, Japan

Transplantation of neural stem/progenitor cells (NS/PCs) promotes functional recovery after spinal cord injury (SCI); however, few studies have examined the optimal site of NS/PC transplantation in the spinal cord. The purpose of this study was to determine the optimal transplantation site of NS/PCs for the treatment of SCI. Wild-type mice were generated with contusive SCI at the T10 level, and NS/PCs were derived from fetal transgenic mice. These NS/PCs ubiquitously expressed ffLuc-cp156 protein (Venus and luciferase fusion protein) and so could be detected by in vivo bioluminescence imaging 9 days postinjury. NS/PCs (low: 250,000 cells per mouse; high: 1 million cells per mouse) were grafted into the spinal cord at the lesion epicenter (E) or at rostral and caudal (RC) sites. Phosphate-buffered saline was injected into E as a control. Motor functional recovery was better in each of the transplantation groups (E-Low, E-High, RC-Low, and RC-High) than in the control group. The photon counts of the grafted NS/PCs were similar in each of the four transplantation groups, suggesting that the survival of NS/PCs was fairly uniform when more than a certain threshold number of cells were transplanted. Quantitative RT-PCR analyses demonstrated that brain-derived neurotropic factor expression was higher in the RC segment than in the E segment, and this may underlie why NS/PCs more readily differentiated into neurons than into astrocytes in the RC group. The location of the transplantation site did not affect the area of spared fibers, angiogenesis, or the expression of any other mediators. These findings indicated that the microenvironments of the E and RC sites are able to support NS/PCs transplanted during the subacute phase of SCI similarly. Optimally, a certain threshold number of NS/PCs should be grafted into the E segment to avoid damaging sites adjacent to the lesion during the injection procedure.

Key words: Spinal cord injury (SCI); Neural stem/progenitor cells (NS/PCs); Cell transplantation; Transplantation site; Subacute phase

Received April 7, 2013; final acceptance August 12, 2013. Online prepub date: August 30, 2013.
Address correspondence to Masaya Nakamura, Department of Orthopedics Surgery, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan. Tel: +81-3-5363-3812; Fax: +81-3-3353-6597; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Hideyuki Okano, Department of Physiology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan. Tel: +81-3-5363-3747; Fax: +81-3-3357-5445; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1465-1470, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X676213
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Brief Communication

Comparison of Olfactory Bulbar and Mucosal Cultures in a Rat Rhizotomy Model

Ahmed Ibrahim,* Daqing Li,* Andrew Collins,* Pawel Tabakow,† Geoffrey Raisman,*‡ and Ying Li*

*Spinal Repair Unit, Department of Brain Repair and Rehabilitation, UCL Institute of Neurology, London, UK
†Department of Neurosurgery, Wroclaw Medical University, Wroclaw, Poland
‡UCL Department of Cell and Developmental Biology, London, UK

In an ongoing clinical trial, a spinal injured patient who received a transplant of autologous cells cultured from the olfactory bulb is showing greater functional benefit than three previous patients with transplants of mucosal origin. Previous laboratory studies of transplantation into rat spinal cord injuries show that the superior reparative benefits of bulbar over mucosal cultures are associated with regeneration of severed corticospinal tract fibers over a bridge of olfactory ensheathing cells (OECs) formed across the injury site. In a rat rhizotomy paradigm, we reported that transplantation of bulbar cell cultures also enables severed axons of the C6–T1 dorsal roots to regenerate across a bridge of OECs into the spinal cord and restore electrophysiological transmission and forepaw grasping during a climbing test. We now report a repeat of the same rhizotomy procedure in 25 rats receiving cells cultured from olfactory mucosal biopsies. In no case did the transplanted cells form a bridging pathway. No axons crossed from the severed roots to the spinal cord, and there was no restoration of forepaw grasping. This suggests that the superior clinical benefit in the patient receiving bulbar cell transplants is due to regeneration of severed fibers across the injury site, and this correlates with imaging and the pattern of functional recovery. Using present culture protocols, the yield of OECs from bulbar biopsies is around 50%, but that from mucosal biopsies is less than 5%. Improving the yield of OECs from mucosal biopsies might avoid the necessity for the intracranial approach to obtain bulbar cells.

Key words: Transplants; Spinal cord injury; Regeneration; Dorsal roots; Olfactory ensheathing cells (OECs)

Received July 4, 2013; final acceptance December 9, 2013. Online prepub date: December 30, 2013.
Address correspondence to Professor Geoffrey Raisman FRS, Chair of Neural Regeneration, Spinal Repair Unit, UCL Institute of Neurology, Queen Square, London WC1N 3BG, UK. Tel: +44 020 7676 2172 (direct), switchboard: +44 020 7837 3611, ext. 84477; Fax: +44 020 7676 2174; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it