Cell Transplantation 23(6) Abstracts

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Cell Transplantation, Vol. 23, pp. 691-701, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X663587
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Enhanced Bone-Forming Activity of Side Population Cells in the Periodontal Ligament

Tadashi Ninomiya,* Toru Hiraga,† Akihiro Hosoya,† Kiyoshi Ohnuma,‡ Yuzuru Ito,§ Masafumi Takahashi,¶ Susumu Ito,** Makoto Asashima,‡§ and Hiroaki Nakamura†

*Division of Hard Tissue Research, Institute for Oral Science, Matsumoto Dental University, Shiojiri, Nagano, Japan
†Department of Histology and Cell Biology, Matsumoto Dental University, Shiojiri, Nagano, Japan
‡Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro-ku, Tokyo, Japan
§Research Center for Stem Cell Engineering (SCRC), National Institute of Advanced Industrial Science and Technology AIST), Higashi, Tsukuba, Ibaraki, Japan
¶Division of Bioimaging Sciences, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan
**Division of Instrumental Analysis, Research Center for Human and Environmental Sciences, Shinshu University, Matsumoto, Nagano, Japan

Regeneration of alveolar bone is critical for the successful treatment of periodontal diseases. The periodontal ligament (PDL) has been widely investigated as a source of cells for the regeneration of periodontal tissues. In the present study where we attempted to develop an effective strategy for alveolar bone regeneration, we examined the osteogenic potential of side population (SP) cells, a stem cell-containing population that has been shown to be highly abundant in several kinds of tissues, in PDL cells. Isolated SP cells from the rat PDL exhibited a superior ability to differentiate into osteoblastic cells compared with non-SP (NSP) and unsorted PDL cells in vitro. The mRNA expressions of osteoblast markers and bone morphogenetic protein (BMP) 2 were significantly upregulated in SP cells and were further increased by osteogenic induction. To examine the bone-forming activity of SP cells in vivo, PDL SP cells isolated from green fluorescent protein (GFP)-transgenic rats were transplanted with hydroxyapatite (HA) disks into wild-type animals. SP cells exhibited a high ability to induce the mineralized matrix compared with NSP and unsorted PDL cells. At 12 weeks after the implantation, some of the pores in the HA disks with SP cells were filled with mineralized matrices, which were positive for bone matrix proteins, such as osteopontin, bone sialoprotein, and osteocalcin. Furthermore, osteoblast- and osteocyte-like cells on and in the bone-like mineralized matrices were GFP positive, suggesting that the matrices were directly formed by the transplanted cells. These results suggest that PDL SP cells possess enhanced osteogenic potential and could be a potential source for cell-based regenerative therapy for alveolar bone.

Keywords: Side population (SP) cells; Periodontal ligament (PDL) cells; Osteogenesis; Bone regeneration

Received June 27, 2012; final acceptance January 25, 2013. Online prepub date: February 5, 2013.
Address correspondence to Tadashi Ninomiya, Ph.D., Division of Hard Tissue Research, Institute for Oral Science, Matsumoto Dental University, 1780 Gobara Hirooka, Shiojiri, Nagano 399-0781, Japan. Tel: +81-263-51-2225; Fax: +81-263-51-2223; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 703-714, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X664577
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Combination Cell Therapy Using Mesenchymal Stem Cells and Regulatory T-Cells Provides a Synergistic Immunomodulatory Effect Associated With Reciprocal Regulation of Th1/Th2 and Th17/Treg Cells in a Murine Acute Graft-Versus-Host Disease Model

Jung-Yeon Lim,* Min-Jung Park,† Keon-Il Im,* Nayoun Kim,* Eun-Joo Jeon,* Eun-Jung Kim,* Mi-La Cho,† and Seok-Goo Cho*‡

*Laboratory of Immune Regulation, Convergent Research Consortium for Immunologic Disease, Seoul St. Mary’s Hospital, The Catholic University of Korea College of Medicine, Seoul, Korea
†Rheumatism Research Center, Catholic Institutes of Medical Science, Seoul St. Mary’s Hospital, The Catholic University of Korea College of Medicine, Seoul, Korea
‡Catholic Blood and Marrow Transplantation Center, Seoul St. Mary’s Hospital, The Catholic University of Korea College of Medicine, Seoul, Korea

Mesenchymal stem cells (MSCs) have been considered to be an ideal cellular source for graft-versus-host disease (GVHD) treatment due to their unique properties, including tissue repair and major histocompatibility complex (MHC)-unmatched immunosuppression. However, preclinical and clinical data have suggested that the immunomodulatory activity of MSCs is not as effective as previously expected. This study was performed to investigate whether the immunomodulatory capacity of MSCs could be enhanced by combination infusion of regulatory T (Treg) cells to prevent acute GVHD (aGVHD) following MHC-mismatched bone marrow transplantation (BMT). For GVHD induction, lethally irradiated BALB/c (H-2d) mice were transplanted with bone marrow cells (BMCs) and spleen cells of C57BL/6 (H-2b) mice. Recipients were injected with cultured recipient-derived MSCs, Treg cells, or MSCs plus Treg cells (BMT + day 0, 4). Systemic infusion of MSCs plus Treg cells improved clinicopathological manifestations and survival in the aGVHD model. Culture of MSCs plus Treg cells increased the population of Foxp3+ Treg cells and suppressed alloreactive T-cell proliferation in vitro. These therapeutic effects were associated with more rapid expansion of donor-type CD4+CD25+Foxp3+ Treg cells and CD4+IL-4+ type 2 T-helper (Th2) cells in the early posttransplant period. Furthermore, MSCs plus Treg cells regulated CD4+IL-17+ Th17 cells, as well as CD4+IFN-γ+ Th1 cells. These data suggest that the combination therapy with MSCs plus Treg cells may have cooperative effects in enhancing the immunomodulatory activity of MSCs and Treg cells in aGVHD. This may lead to development of new therapeutic approaches to clinical allogeneic hematopoietic cell transplantation.

Key words: Allogeneic bone marrow transplantation; Graft-versus-host disease (GVHD); Mesenchymal stem cells (MSCs); Regulatory T-cell

Received July 1, 2012; final acceptance February 7, 2013. Online prepub date: February 26, 2013.
Address correspondence to Professor Seok-Goo Cho, M.D., Ph.D,. Laboratory of Immune Regulation, Convergent Research Consortium for Immunologic Disease, Department of Hematology, Catholic Blood and Marrow Transplantation Center, Seoul St. Mary’s Hospital, The Catholic University of Korea College of Medicine, 222, Banpo-daero, Seocho-gu, Seoul 137-701, Korea. Tel: +82 2 2258 6052; Fax: +82-2-599-3589; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 715-727, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X669770
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Combined Effects of Connective Tissue Growth Factor-Modified Bone Marrow-Derived Mesenchymal Stem Cells and NaOH-Treated PLGA Scaffolds on the Repair of Articular Cartilage Defect in Rabbits

Songsong Zhu,*† Bi Zhang,* Cheng Man,* Yongqing Ma,* Xianwen Liu,* and Jing Hu*†

*State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan Province, People’s Republic of China
†West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan Province, People’s Republic of China

In cartilage tissue engineering using stem cells, it is important to stimulate proliferation and control the differentiation of stem cells to specific lineages. Here we reported a combined technique for articular cartilage repair, consisting of bone marrow mesenchymal stem cells (BMMSCs) transfected with connective tissue growth factor (CTGF) gene and NaOH-treated poly(lactic-co-glycolic) acid (PLGA) scaffolds. In the present study, BMMSCs or CTGF-modified BMMSCs seeded on PLGA or NaOH-treated PLGA scaffolds were incubated in vitro and NaOH-treated PLGA significantly stimulated proliferation of BMMSCs, while CTGF gene transfer promoted chondrogenic differentiation. The effects of the composite on the repair of cartilage defects were evaluated in rabbit knee joints in vivo. Full-thickness cartilage defects (diameter: 5 mm; depth: 3 mm) were created unilaterally in the patellar groove. Defects were either left empty (n = 18) or implanted with BMMSCs/PLGA (n = 18), BMMSCs/NaOH-treated PLGA (n = 18), or CTGF-modified BMMSCs/NaOH-treated PLGA (n = 18). The defect area was examined grossly, histologically, and mechanically at 6, 12, and 24 weeks postoperatively. Implanted cells were tracked using adeno-LacZ labeling at 6 weeks after implantation. Overall, the CTGF-modified BMMSCs/NaOH-treated PLGA group showed successful hyaline-like cartilage regeneration similar to normal cartilage, which was superior to the other groups using gross examination, qualitative and quantitative histology, and mechanical assessment. The in vivo viability of the implanted cells was demonstrated by their retention for 6 weeks after implantation. These findings suggested that a combination of CTGFmodified BMMSCs and NaOH-treated PLGA may be an alternative treatment for large osteochondral defects in high-loading sites.

Key words: Mesenchymal stem cells; Articular cartilage; Defect and repair; Connective tissue growth factor (CTGF); Poly(lactic-co-glycolic) acid (PLGA)

Received March 9, 2011; final acceptance May 4, 2012. Online prepub date: July 11, 2013.
Address correspondence to Jing Hu, D.D.S., M.D., Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, No. 14, 3rd Section of Renminnan Road, Chengdu, Sichuan Province, 610041, People’s Republic of China. Tel: +86-28-85502334; Fax: +86-28-85503552; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 729-745, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X664540
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Autologous Bone Marrow-Derived Cell Therapy Combined With Physical Therapy Induces Functional Improvement in Chronic Spinal Cord Injury Patients

Wael Abo El-kheir,* Hala Gabr,† Mohamed Reda Awad,‡ Osama Ghannam,§ Yousef Barakat,§ Haithem A. M. A. Farghali,¶ Zeinab M. El Maadawi,# Ibrahim Ewes,§ and Hatem E. Sabaawy†**

*Department of Immunology, Military Medical Academy, Cairo, Egypt
†Department of Hematology, Faculty of Medicine, Cairo University, Cairo, Egypt
‡Department of Physical Medicine and Rehabilitation, Al-Azhar University, Cairo, Egypt
§Neurosurgery, Faculty of Medicine, Al-Azhar University, Cairo, Egypt
¶Department of Veterinary Surgery, Anesthesiology and Radiology, Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt
#Department of Histology, Faculty of Medicine, Cairo University, Cairo, Egypt
**Regenerative and Molecular Medicine Program, Department of Medicine, Rutgers–Robert Wood Johnson Medical School, and Rutgers–Cancer Institute of New Jersey, New Brunswick, NJ, USA

Spinal cord injuries (SCI) cause sensory loss and motor paralysis. They are normally treated with physical therapy, but most patients fail to recover due to limited neural regeneration. Here we describe a strategy in which treatment with autologous adherent bone marrow cells is combined with physical therapy to improve motor and sensory functions in early stage chronic SCI patients. In a phase I/II controlled single-blind clinical trial (clinicaltrials.gov identifier: NCT00816803), 70 chronic cervical and thoracic SCI patients with injury durations of at least 12 months were treated with either intrathecal injection(s) of autologous adherent bone marrow cells combined with physical therapy or with physical therapy alone. Patients were evaluated with clinical and neurological examinations using the American Spinal Injury Association (ASIA) Impairment Scale (AIS), electrophysiological somatosensory-evoked potential, magnetic resonance imaging (MRI), and functional independence measurements. Chronic cervical and thoracic SCI patients (15 AIS A and 35 AIS B) treated with autologous adherent bone marrow cells combined with physical therapy showed functional improvements over patients in the control group (10 AIS A and 10 AIS B) treated with physical therapy alone, and there were no long-term cell therapy-related side effects. At 18 months posttreatment, 23 of the 50 cell therapy-treated cases (46%) showed sustained functional improvement. Compared to those patients with cervical injuries, a higher rate of functional improvement was achieved in thoracic SCI patients with shorter durations of injury and smaller cord lesions. Therefore, when combined with physical therapy, autologous adherent bone marrow cell therapy appears to be a safe and promising therapy for patients with chronic SCI of traumatic origin. Randomized controlled multicenter trials are warranted.

Key words: Spinal cord injury (SCI); Cell therapy; Physical therapy; Motor and sensory function; Clinical trial

Received May 10, 2012; final acceptance January 29, 2013. Online prepub date: April 12, 2013.
Address correspondence to Hatem E. Sabaawy, M.D., Ph.D., Regenerative and Molecular Medicine Program, CINJ/Robert Wood Johnson Medical School, 195 Little Albany Street, New Brunswick, NJ 08903. Tel: +1-732-235-8081; Fax: +1-732-235-8681; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 747-760, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X669761
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Wnt1 From Cochlear Schwann Cells Enhances Neuronal Differentiation of Transplanted Neural Stem Cells in a Rat Spiral Ganglion Neuron Degeneration Model

Ya He,*†1 Peng-zhi Zhang,*1 Dong Sun,‡1 Wen-juan Mi,* Xin-yi Zhang,* Yong Cui,* Xing-wang Jiang,* Xiao-bo Mao,* and Jian-hua Qiu*

*Department of Otolaryngology–Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, Xi’an, China
†Department of Otolaryngology, the 153th Hospital of PLA, Zhengzhou, Henan, China
‡Orthopedic Department, Southwest Hospital, Third Military Medical University, Chongqing, China

Although neural stem cell (NSC) transplantation is widely expected to become a therapy for nervous system degenerative diseases and injuries, the low neuronal differentiation rate of NSCs transplanted into the inner ear is a major obstacle for the successful treatment of spiral ganglion neuron (SGN) degeneration. In this study, we validated whether the local microenvironment influences the neuronal differentiation of transplanted NSCs in the inner ear. Using a rat SGN degeneration model, we demonstrated that transplanted NSCs were more likely to differentiate into microtubule-associated protein 2 (MAP2)-positive neurons in SGN-degenerated cochleae than in control cochleae. Using real-time quantitative PCR and an immunofluorescence assay, we also proved that the expression of Wnt1 (a ligand of Wnt signaling) increases significantly in Schwann cells in the SGN-degenerated cochlea. We further verified that NSC cultures express receptors and signaling components for Wnts. Based on these expression patterns, we hypothesized that Schwann cell-derived Wnt1 and Wnt signaling might be involved in the regulation of the neuronal differentiation of transplanted NSCs. We verified our hypothesis in vitro using a coculture system. We transduced a lentiviral vector expressing Wnt1 into cochlear Schwann cell cultures and cocultured them with NSC cultures. The coculture with Wnt1-expressing Schwann cells resulted in a significant increase in the percentage of NSCs that differentiated into MAP2-positive neurons, whereas this differentiation-enhancing effect was prevented by Dkk1 (an inhibitor of the Wnt signaling pathway). These results suggested that Wnt1 derived from cochlear Schwann cells enhanced the neuronal differentiation of transplanted NSCs through Wnt signaling pathway activation. Alterations of the microenvironment deserve detailed investigation because they may help us to conceive effective strategies to overcome the barrier of the low differentiation rate of transplanted NSCs.

Key words: Neural stem cells (NSCs); Transplantation; Inner ear; Schwann cells; Wnt signaling pathway

Received December 6, 2011; final acceptance June 8, 2012. Online prepub date: June 27, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Jian-Hua Qiu, Department of Otolaryngology–Head and Neck Surgery, Xijing Hospital, Fourth Military Medical University, No. 15 Changle Western Road, Xi’an 710032, China. Tel: +86 029 84775381; Fax: +86 029 83224749; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it