Cell Transplantation 25(7) Abstracts

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Cell Transplantation, Vol. 25, pp. 1219-1225, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689947
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Review

Digital Image Analysis to Assess Quantity and Morphological Quality of Isolated Pancreatic Islets

Ling-jia Wang* and Dixon B. Kaufman†

*Department of Surgery, University of Illinois at Chicago, Chicago, IL, USA
†Department of Surgery, University of Wisconsin, Madison, WI, USA

Quantity and quality assessment of human pancreatic islets are essential processes to define a safe and potent quality product used for clinical transplantation. The conventional method of manual assessment has been used in the field for longer than two decades. The high degree of variability in product quantity and lack of archival imaging records of the product for verification are two major disadvantages of using the manual method for quantity and quality assessment of human pancreatic islets. Investigators have developed promising new methods for technical improvement. In this study, we briefly review the published methods and highlight the advantages of digital imaging analysis (DIA) when compared to the manual method. The application of DIA reduces measurement variability and increases the precision of islet equivalent (IEQ) determination for batch analysis. It produces images that can be archived for retrospective analysis and validation, and the data can be transmitted electronically for off-site analysis. These features are important for quality pancreatic islet assessment and are consistent with FDA requirements of current good manufacturing practice for clinical islet transplantation.

Key words: Islets; Digital Imaging; Manufacturing

Received August 30, 2015; final acceptance March 15, 2016. Online prepub date: November 25, 2015.
Address correspondence to Dixon B. Kaufman, M.D., Ph.D. University of Wisconsin–Madison, Department of Surgery, Division of Transplantation, H5/701, Clinical Sciences Center, 600 Highland Ave., Madison, WI 53792, USA Tel: +608-265-6471; Fax: +608-262-6280; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1227-1236, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691286
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Review

Instant Blood-Mediated Inflammatory Reaction in Hepatocyte Transplantation: Current Status and Future Perspectives

Charlotte A. Lee,* Anil Dhawan,†1 Richard A. Smith,‡ Ragai R. Mitry,* and Emer Fitzpatrick*†1

*Institute of Liver Studies, King’s College London, School of Life Sciences and Medicine, King’s College Hospital, London, UK
†Paediatric Liver, GI and Nutrition Centre, King’s College Hospital, London, UK
‡Protein Therapeutics Laboratory, MRC Centre for Transplantation, Faculty of Life Sciences and Medicine, King’s College London at Guy’s Hospital, London, UK

Hepatocyte transplantation (HT) is emerging as a promising alternative to orthotopic liver transplantation (OLT) in patients with certain liver-based metabolic disease and acute liver failure. Hepatocytes are generally infused into the portal venous system, from which they migrate into the liver cell plates of the native organ. One of the major hurdles to the sustained success of this therapy is early cell loss, with up to 70% of hepatocytes lost immediately following infusion. This is largely thought to be due to the instant blood-mediated inflammatory reaction (IBMIR), resulting in the activation of complement and coagulation pathways. Transplanted hepatocytes produce and release tissue factor (TF), which activates the coagulation pathway, leading to the formation of thrombin and fibrin clots. Thrombin can further activate a number of complement proteins, leading to the activation of the membrane attack complex (MAC) and subsequent hepatocyte cell death. Inflammatory cells including granulocytes, monocytes, Kupffer cells, and natural killer (NK) cells have been shown to cluster around transplanted hepatocytes, leading to their rapid clearance shortly after transplantation. Current research aims to improve cell engraftment and prevent early cell loss. This has been proven successful in vitro using pharmacological interventions such as melagatran, low-molecular-weight dextran sulphate, and N-acetylcysteine (NAC). Effective inhibition of IBMIR would significantly improve hepatocyte engraftment, proliferation, and function, providing successful treatment for patients with liver-based metabolic diseases.

Key words: Hepatocyte transplantation (HT); Complement; Coagulation; Innate immunity; Thromboinflammation

Received December 4, 2015; final acceptance March 18, 2016. Online prepub date: March 18, 2016.
1Joint senior authors.
Address correspondence to Dr. Emer Fitzpatrick, Institute of Liver Studies, King’s College London, School of Life Sciences and Medicine, King’s College Hospital, Denmark Hill, London SE5 9RS, UK. Tel: +44-20-3299-1066; Fax: +44-20-3299-4375; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1237-1246, 2016
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DOI: http://dx.doi.org/10.3727/096368915X689541
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Hepatocyte-Like Cells Derived From Mouse Induced Pluripotent Stem Cells Produce Functional Coagulation Factor IX in a Hemophilia B Mouse Model

Yao-Ming Wu,*1 Yu-Jen Huang,*†1 Poda Chen,‡ Yu-Chen Hsu,§ Shu-Wha Lin,§ Hong-Shiee Lai,* and Hsuan-Shu Lee†¶

*Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan
†Institute of Biotechnology, National Taiwan University, Taipei, Taiwan
‡Department of Surgery, National Taiwan University Hospital Yun-Lin Branch, Yunlin, Taiwan
§Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University College of Medicine, Taipei, Taiwan
¶Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan

Hemophilia B (HB) is an inherited deficiency in coagulation factor IX (FIX) that leads to prolonged bleeding after injury. Although hepatocyte transplantation has been demonstrated to be an effective therapeutic strategy for HB, the quality and sources of hepatocytes still limit their application. Recently, stem cells were proposed as an alternative source of donor cells for cell-based therapy. Much research has been devoted to the properties of stem cells that can be differentiated into functional hepatocytes, thereby providing a new cell source for cell-based therapy. Induced pluripotent stem cells (iPSCs) represent a renewable source of hepatocytes for cell-based therapy; these cells exhibit pluripotency and differentiation ability and can be derived from somatic cells. These iPSCs are highly similar to embryonic stem cells (ESCs). We hypothesized that hepatocyte-like cells derived from iPSCs would have therapeutic efficiency in a mouse model of HB. To test this hypothesis, we differentiated iPSCs toward hepatocytes by stepwise protocol and then transplanted these cells into HB mice. We found that these cells shared many characteristics with hepatocytes, such as albumin synthesis, metabolic capacity, glycogen storage, and ureagenesis. Moreover, iPSC-derived hepatocyte transplantation led to increased coagulation factor IX activity, improved thrombus generation, and better hemostasis parameters, and the transferred cells were localized in the liver in recipient HB mice. In conclusion, our results clearly demonstrate that hepatocyte-like cells derived from iPSCs represent a potential cell source for cell-based therapy in the treatment of HB.

Key words: Hemophilia B (HB); Factor IX (FIX); Induced pluripotent stem cells (iPSCs); Transplantation

Received May 1, 2015; final acceptance March 2, 2016. Online prepub date: September 21, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Prof. Hsuan-Shu Lee, Department of Internal Medicine, National Taiwan University Hospital, No. 7 Chung-Shan South Road, Taipei 100, Taiwan. Tel: 886-2-23123456, #65044; Fax: +886-2-33223476; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Prof. Hong-Shiee Lai, Department of Surgery, National Taiwan University Hospital, No. 7 Chung-Shan South Road, Taipei 100, Taiwan. Tel: +886-2-23123456, #65118; Fax: +886-2-33223476; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1247-1257, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689424
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Dimethyl Sulfoxide (DMSO) Increases Percentage of CXCR4+ Hematopoietic Stem/Progenitor Cells, Their Responsiveness to an SDF-1 Gradient, Homing Capacities, and Survival

Danuta Jarocha,* Ewa Zuba-Surma,† and Marcin Majka*

*Department of Transplantation, Jagiellonian University Medical College, Cracow, Poland
†Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland

Cryopreservation of bone marrow (BM), mobilized peripheral blood (mPB), and cord blood (CB) hematopoietic stem/progenitor cells (HSPCs) is a routine procedure before transplantation. The most commonly used cryoprotectant for HSPCs is dimethyl sulfoxide (DMSO). The objective of this study was to evaluate the influence of DMSO on surface receptor expression and chemotactic activities of HSPCs. We found that 10 min of incubation of human mononuclear cells (MNCs) with 10% DMSO significantly increases the percentage of CXCR4+, CD38+, and CD34+ cells, resulting in an increase of CD34+, CD34+CXCR4+, and CD34+CXCR4+CD38 subpopulations. Furthermore, DMSO significantly increased chemotactic responsiveness of MNCs and CXCR4+ human hematopoietic Jurkat cell line to a stromal cell-derived factor-1 (SDF-1) gradient. Furthermore, we demonstrated enhanced chemotaxis of human clonogenic progenitor cells to an SDF-1 gradient, which suggests that DMSO directly enhances the chemotactic responsiveness of early human progenitors. DMSO preincubation also caused lower internalization of the CXCR4 receptor. In parallel experiments, we found that approximately 30% more of DMSO-preincubated human CD45+ and CD45+CD34+ cells homed to the mouse BM 24 h after transplantation in comparison to control cells. Finally, we demonstrated considerably higher (25 days) survival of mice transplanted with DMSO-exposed MNCs than those transplanted with the control cells. We show in this study an unexpected beneficial influence of DMSO on HSPC homing and suggest that a short priming with DMSO before transplantation could be considered a new strategy to enhance cell homing and engraftment.

Key words: Hematopoietic stem/progenitor cells (HSPCs); Cryopreservation; Dimethyl sulfoxide (DMSO); SDF-1–CXCR4 axis; Homing

Received July 22, 2015; final acceptance February 19, 2016. Online prepub date: September 4, 2015.
Address correspondence to Marcin Majka, Department of Transplantation, Jagiellonian University Medical College, Wielicka 265, 30-663 Cracow, Poland. Tel: +48-12-659-1593; Fax: +48-12-659-1594; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1259-1264, 2016
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DOI: http://dx.doi.org/10.3727/096368916X690412
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Assessment of Platelet Activation and Immature Platelet Fraction as Predictors of Platelet Engraftment After Hematopoietic Stem Cell Transplantation

Tang-Her Jaing,* Shih-Hsiang Chen,* Yu-Chuan Wen,† Tsung-Yen Chang,* Jing-Long Huang,‡ and Pei-Kwei Tsay§

*Division of Hematology/Oncology, Department of Pediatrics, Chang Gung Children’s Hospital, Chang Gung University, Linkou, Taoyuan, Taiwan
†Department of Nursing, Chang Gung Memorial Hospital, Linkou, Taoyuan, Taiwan
‡Division of Allergy, Asthma, and Rheumatology, Department of Pediatrics, Chang Gung Children’s Hospital, Chang Gung University, Linkou, Taoyuan, Taiwan
§Department of Public Health and Center of Biostatistics, College of Medicine, Chang Gung University, Linkou, Taoyuan, Taiwan

Delayed platelet engraftment is a well-known complication of umbilical cord blood transplantation (CBT). Megakaryocytes derived from cord blood (CB) in vitro are smaller than megakaryocytes derived from bone marrow (BM) in adults. A small megakaryocyte size might contribute to delayed megakaryocytic maturation. This study included 37 patients undergoing hematopoietic stem cell transplantation (HSCT) at Chang Gung Children’s Hospital between July 2011 and June 2013. Blood samples were obtained at different times: preconditioning and post-HSCT days 56 and 97. To test whether platelet activation persists posttransplantation, two commonly used platelet activation marker antibodies, CD62P (P-selectin) and CD42b, were evaluated using whole blood flow cytometry, combining thiazole orange and anti-CD41a staining, to assess reticulated platelets. Serial peripheral blood (PB) samples were obtained posttransplantationfrom patients undergoing CBT (CBT group; n = 15) and mobilized peripheral blood transplantation (PBT group; n = 22). Platelet activation in the postengraftment samples was considerably higher in the PBT group than the CBT group. Moreover, immature platelet fractions (IPF) were higher in the CBT group. Our results emphasize the role of IPF for dynamic prediction of platelet engraftment in CBT.

Key words: Delayed platelet engraftment; Umbilical cord blood transplantation; Megakaryocytes; Platelet activation; Immature platelet fraction (IPF)

Received September 25, 2015; final acceptance March 10, 2016. Online prepub date: January 13, 2016.

Address correspondence to Tang-Her Jaing, M.D., Division of Hematology and Oncology, Department of Pediatrics, Chang Gung Memorial Hospital, Chang Gung University, 5 Fu-Shin Street, Kwei-Shan, Taoyuan, Taiwan. Tel: +886 3328-1200, ext. 8226; Fax: +886 3328-8957; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1265-1276, 2016
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DOI: http://dx.doi.org/10.3727/096368915X688957
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Downregulation of Heme Oxygenase 1 (HO-1) Activity in Hematopoietic Cells Enhances Their Engraftment After Transplantation

Mateusz Adamiak,* Joseph B. Moore IV,† John Zhao,† Ahmed Abdelbaset-Ismail,* Kamil Grubczak,‡ Sylwia Rzeszotek,*§ Marcin Wysoczynski,† and Mariusz Z. Ratajczak*¶

*Stem Cell Institute at James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA
†Institute of Molecular Cardiology, University of Louisville, Louisville, KY, USA
‡Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Bialystok, Poland
§Department of Physiology, Pomeranian Medical University in Szczecin, Szczecin, Poland
¶Department of Regenerative Medicine, Medical University of Warsaw, Warsaw, Poland

Heme oxygenase 1 (HO-1) is an inducible stress-response enzyme that not only catalyzes the degradation of heme (e.g., released from erythrocytes) but also has an important function in various physiological and pathophysiological states associated with cellular stress, such as ischemic/reperfusion injury. HO-1 has a well-documented anti-inflammatory potential, and HO-1 has been reported to have a negative effect on adhesion and migration of neutrophils in acute inflammation in a model of peritonitis. This finding is supported by our recent observation that hematopoietic stem progenitor cells (HSPCs) from HO-1 KO mice are easy mobilizers, since they respond better to peripheral blood chemotactic gradients than wild-type littermates. Based on these findings, we hypothesized that transient inhibition of HO-1 by nontoxic small-molecule inhibitors would enhance migration of HSPCs in response to bone marrow chemoattractants and thereby facilitate their homing. To directly address this issue, we generated several human hematopoietic cell lines in which HO-1 was upregulated or downregulated. We also exposed murine and human BM-derived cells to small-molecule activators and inhibitors of HO-1. Our results indicate that HO-1 is an inhibitor of hematopoietic cell migration in response to crucial BM homing chemoattractants such as stromal-derived factor 1 (SDF-1) and sphingosine-1-phosphate (S1P). Most importantly, our in vitro and in vivo animal experiments demonstrate for the first time that transiently inhibiting HO-1 activity in HSPCs by small-molecule inhibitors improves HSPC engraftment. We propose that this simple and inexpensive strategy could be employed in the clinical setting to improve engraftment of HSPCs, particularly in those situations in which the number of HSPCs available for transplant is limited (e.g., when transplanting umbilical cord blood).

Key words: Stem cell homing; Stromal-derived factor 1 (SDF-1); Sphingosine-1-phosphate (S1P); Heme oxygenase 1 (HO-1); Chemotaxis; Adhesion; Hematopoietic recovery

Received July 31, 2015; final acceptance February 24, 2016. Online prepub date: July 8, 2015.
Address correspondence to Prof. Mariusz Z. Ratajczak, M.D., Ph.D., Stella and Henry Hoenig Endowed Chair, Professor and Director Stem Cell Institute at James Graham Brown Cancer Center, University of Louisville, 500 S. Floyd Street, Rm. 107, Louisville, KY 40202, USA. Tel: +1-502-852-1788; Fax: +1-502-852-3032; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1277-1286, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689631
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Indiscernible Benefit of Double-Unit Umbilical Cord Blood Transplantation in Children: A Single-Center Experience From Hong Kong

Kam Sze Tsang, Alex Wing Kwan Leung, Vincent Lee, Frankie Wai Tsoi Cheng, Matthew Ming Kong Shing, Henry Nga Hin Pong, Ting Fan Leung, Patrick Man Pan Yuen, and Chi Kong Li

Department of Pediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong

Double-unit umbilical cord blood (DU-UCB) may extend the use of UCB transplantation and improve clinical outcomes. Data in the literature show that single-unit dominance happened in a vast majority of recipients, and the mechanism is unknown. We examined the clinical relevance and engraftment kinetics of DU-UCB transplant in 65 consecutive children who underwent unrelated single-unit (n = 25) and double-unit (n = 40) UCB transplantation for various hematological malignancies (n = 45) and nonmalignant disorders (n = 20). Our result showed no discernible benefit to children receiving double-unit transplant over those receiving single-unit transplant when the total nucleated cell (TNC) doses are ≥2.5 × 107/kg, in terms of the hastening of the engraftment of neutrophils and platelets, reduction of nonengraftment, disease recurrence, early mortality, and graft-versus-host disease, despite significantly higher numbers of TNCs in double units. Further analyses demonstrated that the phenomena were not associated with underlying disease, duration of UCB storage, postthaw viability, HLA disparity, ABO incompatibility, gender, or doses of TNCs, CD34+ cells, CD3+ cells, or colony-forming units. Engrafting units in DU-UCB transplants were notably associated with higher CD34+ cell dose. Chimerism studies demonstrated that single-unit dominance started before neutrophil engraftment in DU-UCB transplants. Data from the study suggested no advantage of infusing double-unit UCB, if an adequately dosed single-unit UCB is available. Successful prediction of the dominant graft would optimize algorithms of UCB selection and maximize the long-term engraftment of chosen units.

Key words: Umbilical cord blood; Transplantation; Engraftment; Chimerism; HLA; Graft-versus-host disease (GvHD)

Received June 10, 2015; final acceptance February 23, 2016. Online prepub date: October 1, 2015.
Address correspondence to Dr. Chi Kong Li, Consultant Pediatrician, Department of Pediatrics, Prince of Wales Hospital, 30-32 Ngan Shing Street, Shatin, N.T., Hong Kong. Tel: (+852) 2632 1019; Fax: (+852) 2649 7859; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1287-1297, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688911
E-ISSN 1555-3892
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A Simple, Rapid, and Efficient Method for Isolating Mesenchymal Stem Cells From the Entire Umbilical Cord

Mehdi Banitalebi Dehkordi,* Zahra Madjd,† Morteza Hashemzadeh Chaleshtori,* Reza Meshkani, ‡ Laleh Nikfarjam,§ and Abdol-Mohammad Kajbafzadeh§

*Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran (IRI)
†Oncopathology Research Center and Dep Pathology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran (IRI)
‡Department of Biochemistry, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran (IRI)
§Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Children’s Hospital Medical Center, Tehran University of Medical Sciences, Tehran, Iran (IRI)

Several reports have been published on the isolation, culture, and identification of mesenchymal stem cells (MSCs) from different anatomical regions of the umbilical cord (UC). UC is suitable for standardizing methods of MSC isolation because it is a uniform source with high MSC numbers. Although the UC is considered a medical waste after childbirth, ethical issues for its use must be considered. An increased demand for MSCs in regenerative medicine has made scientists prioritize the development of MSC isolation methods. Several research groups are attempting to provide a large number of high-quality MSCs. In this study, we present a modulated explant/enzyme method (MEEM) to isolate the maximum number of MSCs from the entire UC. This method was established for the isolation of MSCs from different anatomical regions of the UC altogether. We could retrieve 6 to 10 million MSCs during 8 to 10 days of primary culture. After three passages, we could obtain 8–10 × 108 cells in 28–30 days. MSCs isolated by this method express CD73, CD90, CD105, and CD44, but they do not express hematopoietic markers CD34 and CD45 or the endothelial marker CD31. The genes SOX2, OCT4, and NANOG are expressed in isolated MSCs. The capacity of these MSCs to differentiate into adipocytes and osteocytes highlights their application in regenerative medicine. This method is simple, reproducible, and cost efficient. Moreover, this method is suitable for the production of a large number of high-quality MSCs from an UC in less than a month, to be used for cell therapy in an 80-kg person.

Key words: Umbilical cord (UC); Mesenchymal stem cells (MSCs); Regenerative medicine

Received December 14, 2014; final acceptance September 8, 2015. Online prepub date: August 12, 2015.
Address correspondence to Dr. Abdol-Mohammad Kajbafzadeh, No. 62, Dr. Gharib’s Street, Keshavarz Boulevard, Tehran, Iran (IRI) 1419433151. Tel/Fax: +98-21-66565400; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1299-1317, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688975
E-ISSN 1555-3892
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Coencapsulation of Target Effector Cells With Mesenchymal Stem Cells Reduces Pericapsular Fibrosis and Improves Graft Survival in a Xenotransplanted Animal Model

Vijayaganapathy Vaithilingam, Margaret D. M. Evans, Anthony Rowe, Penelope A. Bean, and Bernard E. Tuch1

Biomedical Manufacturing Research Program, CSIRO Manufacturing Flagship, North Ryde, NSW, Australia Pericapsular fibrotic overgrowth (PFO) is a problem that thwarts full implementation of cellular replacement therapies involving encapsulation in an immunoprotective material, such as for the treatment of diabetes. Mesenchymal stem cells (MSCs) have inherent anti-inflammatory properties. We postulated that coencapsulation of MSCs with the target cells would reduce PFO. A hepatoinsulinoma cell line (HUH7) was used to model human target cells and was coencapsulated with either human or mouse MSCs at different ratios in alginate microcapsules. Viability of encapsulated cells was assessed in vitro and xenografted either intraperitoneally or subcutaneously into C57BL/6 mice. Graft retrieval was performed at 3 weeks posttransplantation and assessed for PFO. Coencapsulation of human MSCs (hMSCs) or mouse MSCs (mMSCs) with HUH7 at different ratios did not alter cell viability in vitro. In vivo data from intraperitoneal infusions showed that PFO for HUH7 cells coencapsulated with hMSCs and mMSCs in a ratio of 1:1 was significantly reduced by ~30% and ~35%, respectively, compared to HUH7 encapsulated alone. PFO for HUH7 cells was reduced by ~51% when the ratio of mMSC/HUH7 was increased to 2:1. Implanting the microcapsules subcutaneously rather than intraperitoneally substantially reduced PFO in all treatment groups, which was most significant in the mMSC/HUH7 2:1 group with a ~53% reduction in PFO compared with HUH7 alone. Despite the reduced PFO reaction to the individual microcapsules implanted subcutaneously, all microcapsule treatment groups were contained in a vascularized fibrotic pouch at 3 weeks. The presence of MSCs in microcapsules retrieved from these fibrotic pouches improved graft survival with significantly higher cell viabilities of 83.1 ± 0.6% and 79.1 ± 0.8% seen with microcapsules containing mMSC/HUH7 at 2:1 and 1:1 ratios, respectively, compared to HUH7 alone (51.5 ± 0.7%) transplanted subcutaneously. This study showed that coencapsulation of MSCs with target cells has a dose-dependent effect on reducing PFO and improving graft survival when implanted either intraperitoneally or subcutaneously in a stringent xenotransplantation setting.

Key words: Mesenchymal stem cells (MSCs); Coencapsulation; Xenotransplantation; Pericapsular fibrosis

Received April 29, 2015; final acceptance November 30, 2015. Online prepub date: September 7, 2015.
1Current address: School of Biomedical Science, Discipline Physiology, University of Sydney, Camperdown, NSW, Australia.
Address correspondence to Vijayaganapathy Vaithlingam, CSIRO Manufacturing Flagship, Riverside Corporate Park, 11 Julius Avenue, Room 4.2.02, PO Box 52, North Ryde, NSW-1670, Australia. Tel: +61 2 9490 5134; Fax: +61 2 9490 5483; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1319-1329, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690449
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Cell-Permeable Peptide Blocks TLR4 Signaling and Improves Islet Allograft Survival

Huansheng Dong,*† Yong Zhang,*† Lili Song,*† Do-sung Kim,* Hongju Wu,‡ Lijun Yang,§ Shiwu Li,§ Katherine A. Morgan,* David B. Adams,* and Hongjun Wang*

*Department of Surgery, Medical University of South Carolina, Charleston, SC, USA
†College of Life Sciences, Qingdao Agricultural University, Qingdao, Shandong, China
‡Department of Medicine, Tulane University, New Orleans, LA, USA
§Department of Pathology, University of Florida, Jacksonville, FL, USA

Toll-like receptor 4 (TLR4) activation in pancreatic β cells activates aberrant islet graft cellular pathways and contributes to immune rejection in allogeneic islet transplantation. As an approach to overcoming this problem, we determined the capacity of a 33-amino acid peptide consisting of a protein transduction domain (PTD) from the Hph-1 virus and a fragment of the intracellular domain of TLR4 from the C3H mice (PTD-dnTLR4) to block TLR4 signaling and improve allogeneic islet survival in vitro and after transplantation. The efficacy of PTD-dnTLR4 in blocking TLR4 signaling was assessed in the Raw264.7 macrophage line, in the islets, and the βTC3 cell line. In Raw264.7 cells, preculture with the peptide reduced LPS-induced NF-κB activation and production of proinflammatory cytokines (IL-1β, TNF-α, iNOS, and IL-6). In islets and β cells, preincubation with PTD-dnTLR4 suppressed LPS-induced TNF-α expression via inhibition of NF-κB activation and protected them from stress-induced cell death. In vivo, preincubation of BALB/c (H-2d) islets with PTD-dnTLR4 resulted in significantly longer survival than control islets in a streptozotocin-induced diabetes model (two of seven grafts survived long term >100 days). PTD-dnTLR4-treated grafts exhibited reduced expression of TNF-α and iNOS and reduced macrophage infiltration posttransplant. The data indicate that PTD-dnTLR4 blocked TLR4 signaling in both macrophages and β cells, and prolonged allograft survival at least in part by suppressing inflammation and macrophage infiltration. This strategy for blocking TLR4 activity has potential utilization in the treatment of diseases where excessive TLR4 activation contributes to the pathologic cellular pathways such as islet transplantation.

Key words: Islet transplantation; Diabetes; Toll-like receptor 4 (TLR4); Graft survival, Inflammation

Received August 12, 2015; final acceptance March 21, 2016. Online prepub date: January 13, 2016.
Address correspondence to Hongjun Wang, Ph.D. Department of Surgery, Medical University of South Carolina, BSB 641, 173 Ashley Ave, Charleston, SC 29425, USA. Tel: 843-792-1800; Fax: 843-792-3315; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1331-1341, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688966
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Kidney Versus Islet Allograft Survival After Induction of Mixed Chimerism With Combined Donor Bone Marrow Transplantation

Tetsu Oura,* Dicken S. C. Ko,* Svjetlan Boskovic,* John J. O’Neil,† Vaja Chipashvili,‡ Maria Koulmanda,‡ Kiyohiko Hotta,* Kento Kawai,* Ognjenka Nadazdin,* R. Neal Smith,* A. B. Cosimi,* and Tatsuo Kawai*

*Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
†Janssen Research & Development, Limited Liability Company, Raritan, NJ, USA
‡Department of Medicine, Transplant Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

We have previously reported successful induction of transient mixed chimerism and long-term acceptance of renal allografts in MHC mismatched nonhuman primates. In this study, we attempted to extend this tolerance induction approach to islet allografts. A total of eight recipients underwent MHC mismatched combined islet and bone marrow (BM) transplantation after induction of diabetes by streptozotocin. Three recipients were treated after a nonmyeloablative conditioning regimen that included low-dose total body and thymic irradiation, horse Atgam (ATG), six doses of anti-CD154 monoclonal antibody (mAb), and a 1-month course of cyclosporine (CyA) (Islet A). In Islet B, anti-CD8 mAb was administered in place of CyA. In Islet C, two recipients were treated with Islet B, but without ATG. The results were compared with previously reported results of eight cynomolgus monkeys that received combined kidney and BM transplantation (Kidney A) following the same conditioning regimen used in Islet A. The majority of kidney/BM recipients achieved long-term renal allograft survival after induction of transient chimerism. However, prolonged islet survival was not achieved in similarly conditioned islet/BM recipients (Islet A), despite induction of comparable levels of chimerism. In order to rule out islet allograft loss due toCyA toxicity, three recipients were treated with anti-CD8 mAb in place of CyA. Although these recipients developed significantly superior mixed chimerism and more prolonged islet allograft survival (61, 103, and 113 days), islet function was lost soon after the disappearance of chimerism. In Islet C recipients, neither prolonged chimerism nor islet survival was observed (30 and 40 days). Significant improvement of mixed chimerism induction and islet allograft survival were achieved with a CyA-free regimen that included anti-CD8 mAb. However, unlike the kidney allograft, islet allograft tolerance was not induced with transient chimerism. Induction of more durable mixed chimerism may be necessary for induction of islet allograft tolerance.

Key words: Kidney transplantation; Islet transplantation; Nonhuman primate; Mixed chimerism

Received February 3, 2015; final acceptance November 9, 2015. Online prepub date: September 2, 2015.
Address correspondence to Tatsuo Kawai, M.D., Ph.D., Department of Surgery, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, White 521, 55 Fruit Street, Boston, MA 02114, USA. Tel: +1-617-817-3270; Fax: +1-617-724-3471; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1343-1357, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690378
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Microcarrier-Expanded Neural Progenitor Cells Can Survive, Differentiate, and Innervate Host Neurons Better When Transplanted as Aggregates

Lifeng Qiu,* Yu Ming Lim,† Allen K. Chen,† Shaul Reuveny,† Steve K. W. Oh,† Eng King Tan,‡§ and Li Zeng*§

*Neural Stem Cell Research Lab, Research Department, National Neuroscience Institute, Singapore
†Stem Cell Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A STAR), Singapore
‡Department of Neurology, National Neuroscience Institute, SGH Campus, Singapore
§Neuroscience and Behavioral Disorders Program, DUKE-NUS Graduate Medical School, Singapore

Neuronal progenitor cells (NPCs) derived from human embryonic stem cells (hESCs) are an excellent cell source for transplantation therapy due to their availability and ethical acceptability. However, the traditional method of expansion and differentiation of hESCs into NPCs in monolayer cultures requires a long time, and the cell yield is low. A microcarrier (MC) platform can improve the expansion of hESCs and increase the yield of NPCs. In this study, for the first time, we transplanted microcarrier-expanded hESC-derived NPCs into the striatum of adult NOD-SCID IL2Rgc null mice, either as single cells or as cell aggregates. The recipient mice were perfused, and the in vivo survival, differentiation, and targeted innervation of the transplanted cells were assessed by immunostaining. We found that both the transplanted single NPCs and aggregate NPCs were able to survive 1 month posttransplantation, as revealed by human-specific neural cell adhesion molecule (NCAM) and human nuclear antigen staining. Compared to the single cells, the transplanted cell aggregates showed better survival over a 3-month period. In addition, both the transplanted single NPCs and the aggregate NPCs were able to differentiate into DCX-positive immature neurons and Tuj1-positive neurons in vivo by 1 month posttransplantation. However, only the transplantation of aggregate NPCs was shown to result in mature neurons at 3 months posttransplantation. Furthermore, we found that the cell aggregates were able to send long axons to innervate their targets. Our study provides preclinical evidence that the use of MCs to expand and differentiate hESC-derived NPCs and transplantation of these cells as aggregates produce longer survival in vivo.

Key words: Neural progenitor cells (NPCs); Transplant; Differentiation; Aggregate; Microcarrier (MC)

Received July 21, 2015; final acceptance March 16, 2016. Online prepub date: December 30, 2015.
Address correspondence to Li Zeng, Neural Stem Cell Research Lab, National Neuroscience Institute, 11 Jalan Tan Tock Seng, Singapore 308433. Tel: +65 6357 7515; Fax: +65 6256 9178; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Steve Oh, Stem Cell Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A STAR), Singapore. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1359-1369, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690421
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Relationship of Grafted FGF-2-Overexpressing Neural Stem/Progenitor Cells With the Vasculature in the Cerebral Cortex

Oleg Tsupykov,*† Michiko Kanemitsu,‡ Ekaterina Smozhanik,* Galina Skibo,*† Alexandre G. Dayer,‡§ and Jozsef Z. Kiss‡

*Department of Cytology, Bogomoletz Institute of Physiology, Kyiv, Ukraine
†Cell and Tissue Technologies Department, State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine
‡Department of Basic Neurosciences, University Medical Center, University of Geneva Medical School, Geneva, Switzerland
§Department of Mental Health and Psychiatry, University Hospital of Geneva, Geneva, Switzerland

Neural progenitor cells (NPCs) overexpressing fibroblast growth factor 2 (FGF-2) have the distinct tendency to associate with the vasculature and establish multiple proliferative clusters in the perivascular environment after transplantation into the cerebral cortex. Strikingly, the vascular clusters of progenitor cells give rise to immature neurons after ischemic injury, raising prospects for the formation of ectopic neurogenic niches for repair. We investigated the spatial relationship of perivascular clusters with the host vascular structures. FGF-2-GFPtransduced NPCs were transplanted into the intact somatosensory rat cortex. Confocal microscopic analysis revealed that grafted cells preferentially contacted venules at sites with aquaporin-4-positive astrocytic endfeet and avoided contacts with desmin-positive pericytes. Electron microscopic analysis confirmed that grafted cells preferentially made contact with astroglial endfeet, and only a minority of them reached the endothelial basal lamina. These results provide new insights into the fine structural and anatomical relationship between grafted FGF-2-transduced NPCs and the host vasculature.

Key words: Fibroblast growth factor 2 (FGF-2); Neural stem cells; Transplantation; Somatosensory cortex; Perivascular clusters

Received August 6, 2015; final acceptance March 16, 2016. Online prepub date: January 22, 2016.
Address correspondence to Jozsef Zoltan Kiss, Department of Neurosciences, University Medical Center (CMU), Rue Michel-Servet 1, 1211 Genève 4, Switzerland. Tel: +41223795202; Fax: +41223795402; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1371-1380, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688533
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Optogenetic Stimulation of Neural Grafts Enhances Neurotransmission and Downregulates the Inflammatory Response in Experimental Stroke Model

Marcel M. Daadi,* Jill Q. Klausner,* Bryce Bajar,* Inbal Goshen,† Christopher Lee-Messer,† Soo Yeun Lee,† Mårten C. G. Winge,‡ Charu Ramakrishnan,† Maisie Lo,† Guohua Sun,* Karl Deisseroth,† and Gary K. Steinberg*

*Department of Neurosurgery, Stanford University School of Medicine, Stanford, CA, USA
†Departments of Bioengineering and Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, CA, USA
‡Stanford Stroke Center, Department of Dermatology, Stanford University School of Medicine, Stanford, CA, USA

Compelling evidence suggests that transplantation of neural stem cells (NSCs) from multiple sources ameliorates motor deficits after stroke. However, it is currently unknown to what extent the electrophysiological activity of grafted NSC progeny participates in the improvement of motor deficits and whether excitatory phenotypes of the grafted cells are beneficial or deleterious to sensorimotor performances. To address this question, we used optogenetic tools to drive the excitatory outputs of the grafted NSCs and assess the impact on local circuitry and sensorimotor performance. We genetically engineered NSCs to express the Channelrhodopsin-2 (ChR2), a light-gated cation channel that evokes neuronal depolarization and initiation of action potentials with precise temporal control to light stimulation. To test the function of these cells in a stroke model, rats were subjected to an ischemic stroke and grafted with ChR2-NSCs. The grafted NSCs identified with a human-specific nuclear marker survived in the peri-infarct tissue and coexpressed the ChR2 transgene with the neuronal markers TuJ1 and NeuN. Gene expression analysis in stimulated versus vehicle-treated animals showed a differential upregulation of transcripts involved in neurotransmission, neuronal differentiation, regeneration, axonal guidance, and synaptic plasticity. Interestingly, genes involved in the inflammatory response were significantly downregulated. Behavioral analysis demonstrated that chronic optogenetic stimulation of the ChR2-NSCs enhanced forelimb use on the stroke-affected side and motor activity in an open field test. Together these data suggest that excitatory stimulation of grafted NSCs elicits beneficial effects in experimental stroke model through cell replacement and non-cell replacement, anti-inflammatory/neurotrophic effects.

Key words: Neural stem cells (NSCs); Neural transplantation; Optogenetic stimulation; Sensorimotor behavior; Stroke

Received March 26, 2015; final acceptance September 28, 2015. Online prepub date: June 30, 2015.
Address correspondence to Marcel Daadi, Ph.D., Southwest National Primate Research Center, Texas Biomedical Research Institute, P.O. Box 760549, San Antonio, TX, 78245-0549, USA. Tel: +1-210-258-9210; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1381-1393, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689893
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Dysfunction in Motor Coordination in Neonatal White Matter Injury Model Without Apparent Neuron Loss

Sachiyo Misumi, Yoshitomo Ueda, Ruriko Nishigaki, Mina Suzuki, Akimasa Ishida, Cha-Gyun Jung, and Hideki Hida

Department of Neurophysiology and Brain Science, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan

We made a white matter injury (WMI) model with mild hindlimb dysfunction by right common carotid artery occlusion followed by 6% oxygen for 60 min at postnatal day 3 (P3), in which actively proliferating oligodendrocyte (OL) progenitors are mainly damaged. To know whether this model is appropriate for cell therapy using OL progenitors, the pathological response to mild hypoxia–ischemia (H-I) in neurons and OL lineage cells and myelination failure were investigated along with gene expression analysis. In WMI model rats, coordinated motor function, as assessed by the accelerating rotarod test, was impaired. The dysfunction was accompanied by myelination failure in layers I–IV of the sensorimotor cortex. Although several oligo2-positive OLs stained positive for active caspase 3 in the cortex and white matter at 24 h after H-I, few NeuN-positive neurons were apoptotic. Argyrophil-III staining for damaged neurons revealed no increase in the number of degenerating cells in the model. Moreover, the total number of NeuN-positive neurons in the cortex was comparable to that of controls 7 days later. Retrograde labeling of the corticospinal tract with Fluoro-Gold revealed no significant loss of layer V neurons. In addition, no decrease in the numbers of cortical projecting neurons and layers V–VI neurons in both motor and sensory areas was observed. Interestingly, the numbers of inhibitory GABAergic cells immunoreactive for parvalbumin, calretinin, or somatostatin were preserved in the P26 cortex. Gene expression analysis at P5 revealed 98 upregulated and 65 downregulated genes that may relate to cell survival, myelin loss, and differentiation of OLs. These data suggest that impaired motor coordination was not induced by neuron loss but, rather, myelination failure in layers I–IV. As OL lineage cells are mainly damaged, this WMI model might be useful for cell-based therapy by replacing OL progenitors.

Key words: Preterm infant; Oligodendrocytes; Hindlimb dysfunction; Active caspase 3; Fluoro-Gold

Received August 20, 2015; final acceptance March 02, 2016. Online prepub date: November 11, 2015.
Address correspondence to Hideki Hida, M.D., Ph.D., Department of Neurophysiology and Brain Science, Nagoya City University Graduate School of Medical Sciences, 1 Kawsasumi, Mizuho-ku, Nagoya 467-8601, Japan. Tel: +1-81-52-853-8134; Fax: +1-81-52-842-3069; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1395-1403, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690198
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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The Use of Autologous Schwann Cells to Supplement Sciatic Nerve Repair With a Large Gap: First in Human Experience

Allan D. Levi, S. Shelby Burks, Kim D. Anderson, Marine Dididze, Aisha Khan, and W. Dalton Dietrich

Department of Neurological Surgery and the Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, USA

Insufficient donor nerve graft material in peripheral nerve surgery remains an obstacle for successful long-distance regeneration. Schwann cells (SCs) can be isolated from adult mammalian peripheral nerve biopsies and can be grown in culture and retain their capacity to enhance peripheral nerve regeneration within tubular repair strategies in multiple animal models. Human Schwann cells (hSCs) can be isolated, expanded in number, and retain their ability to promote regeneration and myelinate axons, but have never been tested in a clinical case of peripheral nerve injury. A sural nerve biopsy and peripheral nerve tissue from the traumatized sciatic nerve stumps was obtained after Food and Drug Administration (FDA) and Institutional Review Board (IRB) approval as well as patient consent. The SCs were isolated after enzymatic digestion of the nerve and expanded with the use of heregulin β1 (0.1 μg/ml) and forskolin (15 mM). After two passages the Schwann cell isolates were combined with sural nerve grafts to repair a large sciatic nerve defect (7.5 cm) after a traumatic nerve injury. The sural nerve and the traumatized sciatic nerve ends both served as an excellent source of purified (90% and 97%, respectively) hSCs. Using ultrasound and magnetic resonance imaging (MRI) we were able to determine continuity of the nerve graft repair and the absence of tumor formation. The patient had evidence of proximal sensory recovery and definitive motor recovery distal to the repair in the distribution of the tibialand common peroneal nerve. The patient did experience an improvement in her pain scores over time. The goals of this approach were to determine the safety and clinical feasibility of implementing a new cellular repair strategy. In summary, this approach represents a novel strategy in the treatment of peripheral nerve injury and represents the first reported use of autologous cultured SCs after human peripheral nerve injury.

Key words: Nerve injury; Repair; Sciatic nerve; Sural nerve

Received August 3, 2015; final acceptance March 17, 2016. Online prepub date: November 25, 2015.
Address correspondence Allan D. Levi, Department of Neurological Surgery, Lois Pope Life Center, University of Miami Miller School of Medicine, 1095 NW 14th Terrace (D4-6), Miami, FL 33136, USA. Tel: (305) 243-2088; Fax: (305) 243-3337; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1405-1414, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689730
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Human Multipotent Mesenchymal Stromal Cells in the Treatment of Postoperative Temporal Bone Defect: An Animal Model

Lukas Skoloudik,* Viktor Chrobok,* David Kalfert,* Zuzana Koci,†‡§ Eva Sykova,†‡ Tetyana Chumak,† Jiri Popelar,† Josef Syka,† Jan Laco,¶ Jana Dedková,# Govindan Dayanithi,†**††‡‡ and Stanislav Filip§§

*Department of Otorhinolaryngology and Head and Neck Surgery, University Hospital Hradec Kralové, Charles University in Prague, Faculty of Medicine in Hradec Kralové, Hradec Kralové, Czech Republic
†Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic
‡Department of Neuroscience, 2nd Faculty of Medicine, Charles University in Prague, Prague, Czech Republic
§Bioinova, Ltd., Prague, Czech Republic
¶The Fingerland Department of Pathology, University Hospital Hradec Kralové, Charles University in Prague, Faculty of Medicine in Hradec Kralové, Hradec Kralové, Czech Republic
#Department of Radiology, University Hospital Hradec Kralové, Charles University in Prague, Faculty of Medicine in Hradec Kralové, Hradec Kralové, Czech Republic
**Department of Molecular Neurophysiology, Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic
††Institut National de la Santé et de la Recherche Médicale, Unité de recherche U1198, Université Montpellier, Montpellier, France
‡‡Ecole Pratique des Hautes Etudes-Sorbonne, Paris, France
§§Department of Oncology and Radiotherapy, Charles University in Prague, Faculty of Medicine in Hradec Kralové, Hradec Kralové, Czech Republic

Canal wall down mastoidectomy is one of the most effective treatments for cholesteatoma. However, it results in anatomical changes in the external and middle ear with a negative impact on the patient’s quality of life. To provide complete closure of the mastoid cavity and normalize the anatomy of the middle and external ear, we used human multipotent mesenchymal stromal cells (hMSCs), GMP grade, in a guinea pig model. A method for preparing a biomaterial composed of hMSCs, hydroxyapatite, and tissue glue was developed. Animals from the treated group were implanted with biomaterial composed of hydroxyapatite and hMSCs, while animals in the control group received hydroxyapatite alone. When compared to controls, the group implanted with hMSCs showed a significantly higher ratio of new bone formation (p = 0.00174), as well as a significantly higher volume percentage of new immature bone (p = 0.00166). Our results proved a beneficial effect of hMSCs on temporal bone formation and provided a promising tool to improve the quality of life of patients after canal wall down mastoidectomy by hMSC implantation.

Key words: Temporal bone; Middle ear surgery; Human bone marrow; Human mesenchymal stromal cells (hMSCs); Guinea pig model; Transplantation; Osteogenesis

Received June 25, 2015; final acceptance February 10, 2016. Online prepub date: October 22, 2015.
Address correspondence to Prof. Stanislav Filip, M.D., Ph.D., D.Sc., Department of Oncology and Radiotherapy, Charles University in Prague, Faculty of Medicine in Hradec Kralové, Hradec Kralové, Czech Republic. Tel: +420 495 834 618; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it



Cell Transplantation, Vol. 25, pp. 1415-1422, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689532
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Osteogenic Ability of Canine Adipose-Derived Mesenchymal Stromal Cell Sheets in Relation to Culture Time

Minyong Kuk,*† Yongsun Kim,*† Seung Hoon Lee,*† Wan Hee Kim,† and Oh-Kyeong Kweon*†

*BK21 PLUS Program for Creative Veterinary Science Research, Research Institute for Veterinary Science and College of Veterinary Medicine, Seoul National University, Seoul, Korea
†Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University, Seoul, Korea

Cell sheets could be used for bone regeneration without requiring a scaffold and can be easily produced from autologous mesenchymal stromal cells (MSCs). We compared the osteogenic potential of MSC-derived cell sheets in relation to culture time. Undifferentiated cell sheets (U-CS) and osteogenic differentiated cell sheets (O-CS) were generated using canine adipose-derived MSCs. Undifferentiated cells (UCs) were used as the control. Osteogenic differentiation was assessed by assaying alkaline phosphatase (ALP) activity. Expression of osteogenesis-related genes was evaluated by reverse transcription-polymerase chain reaction at 4, 7, 14, and 21 days after initiation of culture. The calcium content in cells was measured, and the cells were stained with Alizarin red S (ARS). The mRNA expression of transforming growth factor-β in U-CS and O-CS at day 4 was higher than that in UCs (p < 0.05). The level of bone morphogenetic protein 7 mRNA in O-CS increased significantly at day 4 and was significantly higher than that of U-CS at day 7. The mRNA level of runt-related transcription factor-2 in both sheet types increased significantly at 7 days of culture. The mRNA level of ALP in O-CS and U-CS increased significantly at day 7, and ALP activity was highest at days 7 and 14, respectively (p < 0.05). The mRNA level of osteocalcin in U-CS and O-CS increased significantly at day 21. O-CS and U-CS showed negative ARS staining but their calcium contents increased marginally at day 21. The O-CS cells started to aggregate at days 10–12, and only a partial sheet remained at day 21. The upregulation of expression of genes related to osteogenic differentiation, peak in ALP activity, and morphological changes in cell sheets suggest that the optimal time for application of O-CS and U-CS is between 7 and 10 days and after 14 days of culture, respectively.

Key words: Canine; Osteogenesis; Mesenchymal stromal cells (MSCs); Osteogenic cell sheet

Received August 17, 2015; final acceptance March 12, 2016. Online prepub date: September 21, 2015.
Address correspondence to Oh-Kyeong Kweon, Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul 151-742, Korea. Tel: +82-2-880-1248; Fax: +82-2-888-2866; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1423, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X692078
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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ERRATUM

The following was originally published in Volume 18, Number 12, pages 1359-1368, 2009 (DOI: 10.3727/096368909X475329). In the article title the word Marrow was misspelled as Barrow. The correct spelling in the title is now shown below.

Functional Recovery After the Transplantation of Neurally Differentiated Mesenchymal Stem Cells Derived From Bone Marrow in a Rat Model of Spinal Cord Injury

Sung-Rae Cho,* Yong Rae Kim,† Hoi-Sung Kang,‡ Sun Hee Yim,* Chang-il Park,* Yoo Hong Min,§ Bae Hwan Lee,¶ Ji Cheol Shin,* and Jong-Baeck Lim‡

*Department & Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, Seoul, Korea
†Department of Rehabilitation Medicine, Pochun Joongmoon University College of Medicine, Seoul, Korea
‡Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Korea
§Department of Hematology, Yonsei University College of Medicine, Seoul, Korea
¶Department of Physiology, Brain Research Institute and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea

This study was designed to investigate functional recovery after the transplantation of mesenchymal stem cells (MSCs) or neurally differentiated MSCs (NMSCs) derived from bone marrow in a rat model of spinal cord injury (SCI). Sprague-Dawley rats were subjected to incomplete SCI using an NYU impactor to create a free drop contusion at the T9 level. The SCI rats were then classified into three groups; MSCs, NMSCs, and phosphate-buffered saline (PBS)-treated groups. The cells or PBS were administrated 1 week after SCI. Basso-Beattie-Bresnahan (BBB) locomotor rating scores were measured at 1-week intervals for 9 weeks. Somatosensory evoked potentials (SSEPs) and motor evoked potentials (MEPs) were also recorded 8 weeks after transplantation. While transplantation of MSCs led to a clear tendency of motor recovery, NMSC-treated rats had significantly improved BBB scores and showed significantly shortened initial latency, N1 latency, and P1 latency of the SSEPs compared to PBS controls. In addition, 5-bromo-2-deoxyuridine (BrdU)-prelabeled MSCs costained for BrdU and glial fibrillary acidic protein (GFAP) or myelin basic protein (MBP) were found rostrally and caudally 5 mm each from the epicenter of the necrotic cavity 4 weeks after transplantation. These results suggest that neurallydifferentiated cells might be an effective therapeutic source for functional recovery after SCI.

Key words: Spinal cord injury; Transplantation; Mesenchymal stem cells (MSCs); Neural differentiation; Functional recovery

Received September 2, 2008; final acceptance September 15, 2009. Online prepub date: September 28, 2009.
Address correspondence to Jong-Baeck Lim, M.D., Ph.D., Department of Laboratory Medicine, Yonsei University College of Medicine, 134 Shinchon-dong, Seodaemun-gu, Seoul, Korea 120-752. Tel: +82 2 2228-2447; Fax: +82 2 363-2794; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it