Cell Transplantation 25(5) Abstracts

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Cell Transplantation, Vol. 25, pp. 783-796, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690502
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Induced Pluripotent Stem Cells With Six Reprogramming Factors From Prairie Vole, Which Is an Animal Model for Social Behaviors

Masafumi Katayama,*†1 Takashi Hirayama,*‡1 Kengo Horie,* Tohru Kiyono,§ Kenichiro Donai,*† Satoru Takeda,‡ Katsuhiko Nishimori,* and Tomokazu Fukuda*¶

*Graduate School of Agricultural Science, Tohoku University, Sendai, Japan
†Japan Society for the Promotion of Science, Tokyo, Japan
‡Department of Obstetrics and Gynecology, Juntendo University, Tokyo, Japan
§Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo, Japan
¶Ecological Genetics Analysis Section, Center for Environmental Biology and Ecosystem, National Institute for Environmental Studies of Japan, Tsukuba, Japan

Prairie voles show strong pair bonding with their mating partners, and they demonstrate parental behavior toward their infants, indicating that the prairie vole is a unique animal model for analysis of molecular mechanisms of social behavior. Until a recent study, the signaling pathway of oxytocin was thought to be critical for the social behavior of prairie voles, but neuron-specific functional research may be necessary to identify the molecular mechanisms of social behavior. Prairie vole pluripotent stem cells of high quality are essential to elucidate the molecular mechanisms of social behaviors. Generation of high-quality induced pluripotent stem cells (iPSCs) would help to establish a genetically modified prairie vole, including knockout and knock-in models, based on the pluripotency of iPSCs. Thus, we attempted to establish high-quality prairie vole-derived iPSCs (pv-iPSCs) in this study. We constructed a polycistronic reprogramming vector, which included six reprograming factors (Oct3/4, Sox2, Klf4, c-myc, Lin28, and Nanog). Furthermore, we evaluated the effect of six reprogramming factors, which included Oct3/4 with the transactivation domain (TAD) of MyoD. Implantation of the pv-iPSCs into immunodeficient mice caused a teratoma with three germ layers. Furthermore, the established pv-iPSCs tested positive for stem cell markers, including alkaline phosphatase activity (ALP), stage-specific embryonic antigen (SSEA)-1, and dependence on leukemia inhibitory factor (LIF). Our data indicate that our newly established pv-iPSCs may be a useful tool for genetic analysis of social behavior.

Key words: Induced pluripotent stem cells (iPSCs); Prairie vole; Social behavior; Animal model; Transposon

Received October 19, 2015; final acceptance January 29, 2016. Online prepub date: January 15, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Tomokazu Fukuda, Ph.D., Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan. Tel/Fax: +81 22-717-8695; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Katsuhiko Nishimori, Ph.D., Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan. Tel: +81 22-717-8770; Fax: +81 22-717-8773; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 797-809, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690403
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Intracranial Transplantation of Hypoxia-Preconditioned iPSC-Derived Neural Progenitor Cells Alleviates Neuropsychiatric Defects After Traumatic Brain Injury in Juvenile Rats

Zheng Zachory Wei,*†1 Jin Hwan Lee,†1 Yongbo Zhang,* Yan Bing Zhu,* Todd C. Deveau,† Xiaohuan Gu,† Megan M. Winter,† Jimei Li,* Ling Wei,*† and Shan Ping Yu†‡

*Laboratories of Stem Cell Biology and Regenerative Medicine, Department of Neurology, Experimental Research Center and Neurological Disease Center, Beijing Friendship Hospital, Capital Medical University, Beijing, China
†Department of Anesthesiology, Emory University School of Medicine, Atlanta, GA, USA
‡Center for Visual and Neurocognitive Rehabilitation, Atlanta VA Medical Center, Decatur, GA, USA

Traumatic brain injury (TBI) is a common cause of mortality and long-term morbidity in children and adolescents. Posttraumatic stress disorder (PTSD) frequently develops in these patients, leading to a variety of neuropsychiatric syndromes. Currently, few therapeutic strategies are available to treat juveniles with PTSD and other developmental neuropsychiatric disorders. In the present investigation, postnatal day 14 (P14) Wistar rats were subjected to TBI induced by a controlled cortical impact (CCI) (velocity = 3 m/s, depth = 2.0 mm, contact time = 150 ms). This TBI injury resulted in not only cortical damages, but also posttrauma social behavior deficits. Three days after TBI, rats were treated with intracranial transplantation of either mouse iPSC-derived neural progenitor cells under normal culture conditions (N-iPSC-NPCs) or mouse iPSC-derived neural progenitor cells pretreated with hypoxic preconditioning (HP-iPSC-NPCs). Compared to TBI animals that received N-iPSC-NPCs or vehicle treatment, HP-iPSC-NPC-transplanted animals showed a unique benefit of improved performance in social interaction, social novelty, and social transmission of food preference tests. Western blotting showed that HP-iPSC-NPCs expressed significantly higher levels of the social behavior-related genes oxytocin and the oxytocin receptor. Overall, HP-iPSC-NPC transplantation exhibits a great potential as a regenerative therapy to improve neuropsychiatric outcomes after juvenile TBI.

Key words: Induced pluripotent stem cell-derived neural progenitor cells (iPSC-NPCs); Hypoxic preconditioning; Regeneration; Juveniles; Traumatic brain injury (TBI); Posttraumatic disorders

Received December 16, 2015; final acceptance January 28, 2016. Online prepub date: January 13, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Shan Ping Yu, M.D., Ph.D., Department of Anesthesiology, Emory University School of Medicine, 101 Woodruff Circle WMRB Suite 620, Atlanta, GA 30322, USA. Tel: +1-404-727-6300; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 811-827, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690467
E-ISSN 1555-3892
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Melanoma Immunotherapy in Mice Using Genetically Engineered Pluripotent Stem Cells

Mohammad Haque,*1 Jianyong Song,†1 Kristin Fino,* Praneet Sandhu,* Youfei Wang,† Bing Ni,† Deyu Fang,‡ and Jianxun Song*

*Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Hershey, PA, USA
†Institutes of Irradiation/Immunology, The Third Military Medical University, Chongqing, China
‡Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

Adoptive cell transfer (ACT) of antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) is a highly promising treatment for a variety of diseases. Naive or central memory T-cell-derived effector CTLs are optimal populations for ACT-based immunotherapy because these cells have a high proliferative potential, are less prone to apoptosis than terminally differentiated cells, and have the higher ability to respond to homeostatic cytokines. However, such ACT with T-cell persistence is often not feasible due to difficulties in obtaining sufficient cells from patients. Here we present that in vitro differentiated HSCs of engineered PSCs can develop in vivo into tumor Ag-specific naive CTLs, which efficiently suppress melanoma growth. Mouse-induced PSCs (iPSCs) were retrovirally transduced with a construct encoding chicken ovalbumin (OVA)-specific T-cell receptors (TCRs) and survival-related proteins (i.e., BCL-xL and survivin). The gene-transducediPSCs were cultured on the Δ-like ligand 1-expressing OP9 (OP9-DL1) murine stromal cells in the presence of murine recombinant cytokines (rFlt3L and rIL-7) for a week. These iPSC-derived cells were then intravenously adoptively transferred into recipient mice, followed by intraperitoneal injection with an agonist α-Notch 2 antibody and cytokines (rFlt3L and rIL-7). Two weeks later, naive OVA-specific CD8+ T cells were observed in the mouse peripheral lymphatic system, which were responsive to OVA-specific stimulation. Moreover, the mice were resistant to the challenge of B16-OVA melanoma induction. These results indicate that genetically modified stem cells may be used for ACT-based immunotherapy or serve as potential vaccines.

Key words: Experimental immunotherapy; Pluripotent stem cells; Genetic modification; Adoptive transfer; Mouse; T cells

Received October 19, 2015; final acceptance January 25, 2016. Online prepub date: January 15, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Jianxun Song, Ph.D., Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, 500 University Dr., Hershey, PA 17033, USA. Tel: +1-717-531-0003, ext. 287768; Fax: +1-717-531-4600; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 829-848, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689622
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Clinical Trials With Mesenchymal Stem Cells: An Update

Tiziana Squillaro,*† Gianfranco Peluso,† and Umberto Galderisi*‡§

*Department of Experimental Medicine, Biotechnology and Molecular Biology Section, Second University of Naples, Naples, Italy
†Institute of Bioscience and Bioresources, National Research Council CNR, Naples, Italy
Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, Temple University, Philadelphia, PA, USA
§Genome and Stem Cell Center (GENKOK), Erciyes University, Kayseri, Turkey

In the last year, the promising features of mesenchymal stem cells (MSCs), including their regenerative properties and ability to differentiate into diverse cell lineages, have generated great interest among researchers whose work has offered intriguing perspectives on cell-based therapies for various diseases. Currently the most commonly used adult stem cells in regenerative medicine, MSCs, can be isolated from several tissues, exhibit a strong capacity for replication in vitro, and can differentiate into osteoblasts, chondrocytes, and adipocytes. However, heterogeneous procedures for isolating and cultivating MSCs among laboratories have prompted the International Society for Cellular Therapy (ISCT) to issue criteria for identifying unique populations of these cells. Consequently, the isolation of MSCs according to ISCT criteria has produced heterogeneous, nonclonal cultures of stromal cells containing stem cells with different multipotent properties, committed progenitors, and differentiated cells. Though the nature and functions of MSCs remain unclear, nonclonal stromal cultures obtained from bone marrow and other tissues currently serve as sources of putative MSCs for therapeutic purposes, and several findings underscore their effectiveness in treating different diseases. To date, 493 MSC-based clinical trials, either complete or ongoing, appear in the database of the US National Institutes of Health. In the present article, we provide a comprehensive review of MSC-based clinical trials conducted worldwide that scrutinizes biological properties of MSCs, elucidates recent clinical findings and clinical trial phases of investigation, highlights therapeutic effects of MSCs, and identifies principal criticisms of the use of these cells. In particular, we analyze clinical trials using MSCs for representative diseases, including hematological disease, graft-versus-host disease, organ transplantation, diabetes, inflammatory diseases, and diseases in the liver, kidney, and lung, as well as cardiovascular, bone and cartilage, neurological, and autoimmune diseases.

Key words: Mesenchymal stem cells (MSCs); Clinical trials; Immunomodulation; Differentiation; Secretome; Paracrine effects

Received July 10, 2015; final acceptance February 3, 2016. Online prepub date: September 29, 2015.
Address correspondence to Umberto Galderisi, Department of Experimental Medicine, Second University of Naples, via Costantinopoli, 16, 80128, Naples, Italy. Tel: +39 081 566-7585; Fax: +39 081 566-7547; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Gianfranco Peluso, Institute of Bioscience and Bioresources, CNR, via Pietro Castellino 111, 80131 Naples, Italy. Tel: +39 081 613-2280; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 849-861, 2016
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DOI: http://dx.doi.org/10.3727/096368916X690881
E-ISSN 1555-3892
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Review

Muse Cells Provide the Pluripotency of Mesenchymal Stem Cells: Direct Contribution of Muse Cells to Tissue Regeneration

Mari Dezawa

Department of Stem Cell Biology and Histology and Department of Anatomy and Anthropology, Tohoku University Graduate School of Medicine, Sendai, Japan

While mesenchymal stem cells (MSCs) are easily accessible from mesenchymal tissues, such as bone marrow and adipose tissue, they are heterogeneous, and their entire composition is not fully identified. MSCs are not only able to differentiate into osteocytes, chondrocytes, and adipocytes, which belong to the same mesodermal lineage, but they are also able to cross boundaries between mesodermal, ectodermal, and endodermal lineages, and differentiate into neuronal- and hepatocyte-like cells. However, the ratio of such differentiation is not very high, suggesting that only a subpopulation of the MSCs participates in this cross-lineage differentiation phenomenon. We have identified unique cells that we named multilineage-differentiating stress-enduring (Muse) cells that may explain the pluripotent-like properties of MSCs. Muse cells comprise a small percentage of MSCs, are able to generate cells representative of all three germ layers from a single cell, and are nontumorigenic and self-renewable. Importantly, cells other than Muse cells in MSCs do not have these pluripotent-like properties. Muse cells are particularly unique compared with other stem cells in that they efficiently migrate and integrate into damaged tissue when supplied into the bloodstream, and spontaneously differentiate into cells compatible with the homing tissue. Such a repairing action of Muse cells via intravenous injection is recognized in various tissues including the brain, liver, and skin. Therefore, unlike ESCs/iPSCs, Muse cells render induction into the target cell type prior to transplantation unnecessary. They can repair tissues in two simple steps: collection from mesenchymal tissues, such as the bone marrow, and intravenous injection. The impressive regenerative performance of these cells provides a simple, feasible strategy for treating a variety of diseases. This review details the unique characteristics of Muse cells and describes their future application for regenerative medicine.

Key words: Mesenchymal stem cells (MSCs); Pluripotent stem cells; Cell therapy; Liver; Muscle; Stroke

Received October 19, 2015; final acceptance February 1, 2016. Online prepub date: February 15, 2016.
Address correspondence to Mari Dezawa, M.D., Ph.D., Department of Stem Cell Biology and Histology and Department of Anatomy and Anthropology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Sendai 980-8575, Japan. Tel: +81-22-717-8025; Fax: +81-22-717-8030; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 863-882, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690511
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Review

Induction of Neurorestoration From Endogenous Stem Cells

Ji Hea Yu,*† Jung-Hwa Seo,*† Ji Yong Lee,*‡ Min-Young Lee,*‡ and Sung-Rae Cho*†‡§

*Department and Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, Seoul, Korea
†Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, Korea
Yonsei Stem Cell Center, Avison Biomedical Research Center, Seoul, Korea
§Rehabilitation Institute of Neuromuscular Disease, Yonsei University College of Medicine, Seoul, Korea

Neural stem cells (NSCs) persist in the subventricular zone lining the ventricles of the adult brain. The resident stem/progenitor cells can be stimulated in vivo by neurotrophic factors, hematopoietic growth factors, magnetic stimulation, and/or physical exercise. In both animals and humans, the differentiation and survival of neurons arising from the subventricular zone may also be regulated by the trophic factors. Since stem/progenitor cells present in the adult brain and the production of new neurons occurs at specific sites, there is a possibility for the treatment of incurable neurological diseases. It might be feasible to induce neurogenesis, which would be particularly efficacious in the treatment of striatal neurodegenerative conditions such as Huntington’s disease, as well as cerebrovascular diseases such as ischemic stroke and cerebral palsy, conditions that are widely seen in the clinics. Understanding of the molecular control of endogenous NSC activation and progenitor cell mobilization will likely provide many new opportunities as therapeutic strategies. In this review, we focus on endogenous stem/progenitor cell activation that occurs in response to exogenous factors including neurotrophic factors, hematopoietic growth factors, magnetic stimulation, and an enriched environment. Taken together, these findings suggest the possibility that functional brain repair through induced neurorestoration from endogenous stem cells may soon be a clinical reality.

Key words: Neural stem cells (NSCs); Neurotrophic factor; Hematopoietic growth factor; Magnetic stimulation; Enriched environment

Received December 16, 2015; final acceptance January 27, 2016. Online prepub date: January 18, 2016.
Address correspondence to Sung-Rae Cho, M.D., Ph.D., Department and Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, 50 Yonsei-roSeodaemun-gu, Seoul, South Korea, 120-752. Tel: +82 2 2228-3715; Fax: +82 2 363-2795; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 883-891, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689749
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Review

Melatonin as an Antioxidant for Stroke Neuroprotection

Nate Watson,1 Theo Diamandis,1 Chiara Gonzales-Portillo, Stephanny Reyes, and Cesar V. Borlongan

Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA

Melatonin (N-acetyl-5-methoxytryptamine) is a hormone derived from the pineal gland that has a wide range of clinical applications. While melatonin was originally assessed as a hormone specializing in regulation of the normal circadian rhythm in mammals, it now has been shown to be an effective free radical scavenger and antioxidant. Current research has focused on central nervous system (CNS) disorders, stroke in particular, for potential melatonin-based therapeutics. As of now, the realm of potential therapy regimens is focused on three main treatments: exogenously delivered melatonin, pineal gland grafting, and melatonin-mediated stem cell therapy. All therapies contain both costs and benefits, and current research is still focused on finding the best treatment plan. While comprehensive research has been conducted, more research regarding the safety of such therapies is needed in order to transition into the clinical level of testing. Antioxidants such as traditional Chinese medicine, (−)-epigallocatechin-3-gallate (EGCG), and lavender oil, which have been used for thousands of years as treatment, are now gaining recognition as effective melatonin treatment alternatives. This review will further discuss relevant studies assessing melatonin-based therapeutics and provide evidence of other natural melatonin treatment alternatives for the treatment of stroke.

Key words: Melatonin; Pineal gland; Antioxidant; Neuroprotection; Melatonin receptors; Dementia

Received July 31, 2015; final acceptance November 5, 2015. Online prepub date: October 22, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Professor Cesar V. Borlongan, Ph.D., Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida,Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA. Tel: +1-813-974-3154; Fax: +1-813-974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 893-898, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690548
E-ISSN 1555-3892
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Review

Recent Progress in Therapeutic Strategies for Ischemic Stroke

Toru Yamashita and Koji Abe

Department of Neurology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

Possible strategies for treating stroke include neuroprotection in the acute phase of cerebral ischemia and stem cell therapy in the chronic phase of cerebral ischemia. Previously, we have studied the temporal and spatial expression patterns of c-fos, hypoxia inducible factor-1α (HIF-1α), heat shock protein 70 (HSP70), and annexin V after 90 min of transient middle cerebral occlusion in rats and concluded that there is a time window for neuroprotection from 12 to 48 h after ischemia. In addition, we have estimated the neuroprotective effect of glial cell line-derived neurotrophic factor (GDNF) by injecting Sendai viral vector containing the GDNF gene into the postischemic brain. This Sendai virus-mediated gene transfer of GDNF showed a significant neuroprotective effect in the ischemic brain. Additionally, we have administered GDNF and hepatocyte growth factor (HGF) protein into the postischemic rat brain and estimated the infarct size and antiapoptoticand antiautophagic effects. GDNF and HGF significantly reduced infarct size, the number of microtubuleassociated protein 1 light chain 3 (LC3)-positive cells, and the number of terminaldeoxynucleotidyl transferase-mediated dUTP-biotin in situ nick-end labeling (TUNEL)-positive cells, indicating that GDNF and HGF were greatly associated with not only the antiapoptoticeffect but also the antiautophagic effects. Finally, we have previously transplanted undifferentiated iPSCs into the ipsilateral striatum and cortex at 24 h after cerebral ischemia. Histological analysis was performed at 14 and 28 days after cell transplantation, and we found that iPSCs could supply a great number of doublecortin-positive neuroblasts but also formed tridermal teratomain the ischemic brain. Our results suggest that iPSCs have a potential to provide neural cells after ischemic brain injury if tumorigenesis is properly controlled. In the future, we will combine these strategies to develop more effective therapies for the treatment of strokes.

Key words: Multiple molecular penumbra; Neurotrophic factor; Induced pluripotent stem cells (iPSCs); Cell transplantation

Received October 19, 2015; final acceptance January 28, 2016. Online prepub date: January 18, 2016.
Address correspondence to Koji Abe, Department of Neurology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. Tel: +81-86-235-7365; Fax: +81-86-235-7368; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 899-912, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690539
E-ISSN 1555-3892
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Therapeutic Effect of Ligustilide-Stimulated Adipose-Derived Stem Cells in a Mouse Thromboembolic Stroke Model

Kang Chi,* Ru-Huei Fu,*† Yu-Chuen Huang,‡§ Shih-Yin Chen,‡§ Shinn-Zong Lin,*† Pi-Chun Huang,¶ Po-cheng Lin,¶ Fu-Kuei Chang,# and Shih-Ping Liu*,**††

*Center for Neuropsychiatry, China Medical University Hospital, Taichung, Taiwan, China
†Graduate Institute of Immunology, China Medical University, Taichung, Taiwan, China
‡Genetics Center, Department of Medical Research, China Medical University Hospital, Taichung, Taiwan, China
§School of Chinese Medicine, College of Chinese Medicine, China Medical University, Taichung, Taiwan, China
¶Department of Stem Cell Applied Technology, Gwo Xi Stem Cell Applied Technology, Hsinchu, Taiwan, China
#Department of Health Management, College of Medicine, I-Shou University, Kaohsiung City, Taiwan, China
**Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan, China
††Department of Social Work, Asia University, Taichung, Taiwan, China

Stroke is a result of cerebral ischemia that triggers a cascade of both physiological and biochemical events. No effective treatment is available for stroke; however, stem cells have the potential to rescue tissue from the effects of stroke. Adipose-derived stem cells (ADSCs) are an abundant source of adult stem cells; therefore, ADSC therapy can be considered as a future strategy for regenerative medicine. However, more research is required to improve the effectiveness of transplanted ADSCs as a treatment for stroke in the mouse stroke model. Ligustilide, isolated from the herb Angelica sinensis, exhibits a protective effect on neurons and inhibits inflammation. We also demonstrated that ligustilide treatment increases the expression levels of homing factors such as SDF-1 and CXCR4. In the present study, we evaluated the therapeutic effects of ADSC transplantation and ligustilide treatment in a mouse thromboembolic stroke model by behavioral tests, including beam walking, locomotor activity, and rotarod analysis. ADSCs pretreated with ligustilide were transplanted into the brains of stroke mice. The results showed that the therapeutic effect of ADSCs pretreated with ligustilide was better than that of ADSCs without ligustilide pretreatment. There was no difference between the recovery of mice treated by ADSC transplantation combined with subcutaneous ligustilide injection and that of mice treated only with ADSCs. The TUNEL assay showed fewer apoptotic cells in the brains of mice transplanted with ADSCs pretreated with ligustilide as well as in those without pretreatment. In summary, pretreatment of ADSCs with ligustilide improves the therapeutic efficacy of ADSC transplantation. The results of this study will help improve stem cell therapies being developed for future clinical applications.

Key words: Stroke; Adipose-derived stem cells (ADSCs); Ligustilide

Received December 16, 2015; final acceptance January 29, 2016. Online prepub date: January 18, 2016.
Address correspondence to Shih-Ping Liu, Ph.D., Center for Neuropsychiatry, China Medical University Hospital, No. 2, Yuh-Der Road, Taichung, Taiwan 40447, ROC. Tel: +886-4-2205-2121, ext. 7828; Fax: +886-4-2205-2121, ext. 7810; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 913-927, 2016
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DOI: http://dx.doi.org/10.3727/096368915X689785
E-ISSN 1555-3892
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Transferring Xenogenic Mitochondria Provides Neural Protection Against Ischemic Stress in Ischemic Rat Brains

Po-Jui Huang,* Chi-Chung Kuo,*†‡ Hsiu-Chin Lee,* Ching-I Shen,§ Fu-Chou Cheng,¶ Shih-Fang Wu,* Jui-Chih Chang,# Hung-Chuan Pan,**†† Shinn-Zong Lin,‡‡§§¶¶ Chin-San Liu,#,##*** and Hong-Lin Su*

*Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, Taichung, Taiwan
†Department of Neurology, Taichung Tzu Chi Hospital, The Buddhist Tzu Chi Medical Foundation, Taichung, Taiwan
‡School of Medicine, Tzu Chi University, Hualien, Taiwan
§Department of Chemistry, National Chung Hsing University, Taichung, Taiwan
¶Stem Cells Center, Department of Medical Research, Taichung Veteran General Hospital, Taichung, Taiwan
#Vascular and Genomic Center, Changhua Christian Hospital, Changhua, Taiwan
**Department of Neurosurgery, Taichung Veteran General Hospital, Taichung, Taiwan
††Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan
‡‡Center for Neuropsychiatry, China Medical University Hospital, Taichung, Taiwan
§§Department of Neurosurgery, China Medical University Beigang Hospital, Yunlin, Taiwan
¶¶Graduate Institute of Immunology, China Medical University, Taichung, Taiwan
##Department of Neurology, Changhua Christian Hospital, Changhua, Taiwan
***Graduate Institute of Integrated Medicine, College of Chinese Medicine, China Medical University, Taichung, Taiwan

Transferring exogenous mitochondria has therapeutic effects on damaged heart, liver, and lung tissues. Whether this protective effect requires the symbiosis of exogenous mitochondria in host cells remains unknown. Here xenogenic mitochondria derived from a hamster cell line were applied to ischemic rat brains and rat primary cortical neurons. Isolated hamster mitochondria, either through local intracerebral or systemic intra-arterial injection, significantly restored the motor performance of brain-ischemic rats. The brain infarct area and neuronal cell death were both attenuated by the exogenous mitochondria. Although internalized mitochondria could be observed in neurons and astrocytes, the low efficacy of mitochondrial internalization could not completely account for the high rate of rescue of the treated neural cells. We further illustrated that disrupting electron transport or ATPase synthase in mitochondria significantly attenuated the protective effect, suggesting that intact respiratory activity is essential for the mitochondrial potency on neural protection. These results emphasize that nonsymbiotic extracellular mitochondria can provide an effective cell defense against acute injurious ischemic stress in the central nervous system.

Key words: Mitochondria; Ischemic stroke; Mitochondria transfer; Neural protection

Received August 18, 2015; final acceptance January 27, 2015. Online prepub date: November 6, 2015.
Address correspondence to Chin-San Liu, M.D., Ph.D., Changhua Christian Hospital, Department of Vascular and Genomic Center, 135, Nanhsiao Street, Changhua, 50094, Taiwan. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Hong-Lin Su, Ph.D., Department of Life Sciences, National Chung Hsing University, 250, Kuo-Kuang Road, Taichung, 402, Taiwan. Tel: +886-422840416; Fax: +886-422854391; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 929-935, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689758
E-ISSN 1555-3892
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Review

Extracardiac-Lodged Mesenchymal Stromal Cells Propel an Inflammatory Response Against Myocardial Infarction via Paracrine Effects

Yi Peng,* Wei Pan,* Yali Ou,* Weifang Xu,* Sussannah Kaelber,† Cesario V. Borlongan,† Meiqin Sun,* and Guolong Yu*

*Department of Cardiology, Xiangya Hospital, Central Southern University, Changsha, Hunan, China
†Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA

Transplantation of stem cells, including mesenchymal stromal cells (MSCs), improves the recovery of cardiac function after myocardial infarction (MI) in experimental studies using animal models and in patients. However, the improvement of cardiac function following MSC transplantation remains suboptimal in both preclinical and clinical studies. Understanding the mechanism of cell therapy may improve its therapeutic outcomes, but the mode of action mediating stem cell promotion of cardiac repair is complex and not fully understood. Recent studies suggest that the immunomodulatory effects of MSCs on the macrophage M1/M2 subtype transition allow the transplanted stem cells to inhibit inflammation-induced injury and promote cardiac repair in acute MI. However, equally compelling evidence shows that there is poor survival and minimal graft persistence of transplanted MSCs within the infarcted heart tissues, negating the view that graft survival per se is required for the observed high rate and long duration of the transition from proinflammatory M1 to reparative M2 macrophages in the infarcted myocardium. Therefore, we raised a novel hypothesis that the therapeutic effects of MSC transplantation for acute MI depends not primarily on the grafted cells in infarct myocardium, but that MSCs migrating to and being lodged in the extracardiac organs, demonstrating good graft survival and persistence, may render the therapeutic effects in MI. More specifically, MSC transplantation promotes the transition from M1 to M2 in extracardiac organs, such as spleen and bone marrow, and therapeutic effects are conferred to the infarcted myocardium via paracrine effects. In MSC transplantation, the conversion from proinflammatory M1 to anti-inflammatory M2 monocytes may occur remotely from the heart and may serve as one of the major pathways in regulating the dual effects of inflammation. This hypothesis, if proven valid, may represent an important new mechanism of action to be considered for the future of MSC transplantation in the treatment of MI.

Key words: Mesenchymal stromal cells (MSCs); Transplantation; Myocardial infarction (MI); Cardiac repair

Received July 31, 2015; final acceptance November 5, 2015. Online prepub date: October 22, 2015.
Address correspondence to Professor Cesar V. Borlongan, Ph.D., Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida,Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA. Tel: +1-813-974-3154; Fax: +1-813-974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 937-950, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690288
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Review

Mesenchymal Stem Cells and Their Clinical Applications in Osteoarthritis

Yu-Hsun Chang,*† Hwan-Wun Liu,†‡ Kun-Chi Wu,§ and Dah-Ching Ding†¶

*Department of Pediatrics, Buddhist Tzu Chi General Hospital, Hualien, Taiwan
†Institute of Medical Sciences, Tzu Chi University, Hualien, Taiwan
‡Department of Occupational Medicine, Buddhist Tzu Chi General Hospital, Hualien, Taiwan
§Department of Orthopedics, Buddhist Tzu Chi General Hospital, Hualien, Taiwan
¶Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, Hualien, Taiwan

Osteoarthritis is a chronic degenerative joint disorder characterized by articular cartilage destruction and osteophyte formation. Chondrocytes in the matrix have a relatively slow turnover rate, and the tissue itself lacks a blood supply to support repair and remodeling. Researchers have evaluated the effectiveness of stem cell therapy and tissue engineering for treating osteoarthritis. All sources of stem cells, including embryonic, induced pluripotent, fetal, and adult stem cells, have potential use in stem cell therapy, which provides a permanent biological solution. Mesenchymal stem cells (MSCs) isolated from bone marrow, adipose tissue, and umbilical cord show considerable promise for use in cartilage repair. MSCs can be sourced from any or all joint tissues and can modulate the immune response. Additionally, MSCs can directly differentiate into chondrocytes under appropriate signal transduction. They also have immunosuppressive and anti-inflammatory paracrine effects. This article reviews the current clinical applications of MSCs and future directions of research in osteoarthritis.

Key words: Mesenchymal stem cells (MSCs); Osteoarthritis (OA); Regeneration; Chondrocyte; Cartilage

Received August 25, 2015; final acceptance February 1, 2016. Online prepub date: December 18, 2015.
Address correspondence to Dah-Ching Ding, M.D., Ph.D., Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, 707, Chung-Yang Rd., Sec. 3, Hualien 970, Taiwan. Tel: +886-3-856-1825; Fax: +886-3-857-7161; E-mail:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 951-962, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690917
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Autotransplantation of Monkey Ear Perichondrium-Derived Progenitor Cells for Cartilage Reconstruction

Shintaro Kagimoto,* Takanori Takebe,†‡§1 Shinji Kobayashi,†¶ Yuichiro Yabuki,¶ Ayaka Hori,† Koichi Hirotomi,* Taro Mikami,* Toshimasa Uemura,# Jiro Maegawa,*1 and Hideki Taniguchi†‡1

*Department of Plastic and Reconstructive Surgery, Yokohama City University Hospital, Yokohama, Kanagawa, Japan
†Department of Regenerative Medicine, Yokohama City University, Yokohama, Kanagawa, Japan
‡Project leader in the Advanced Medical Research Center, Yokohama, Kanagawa, Japan
§Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA
¶Department of Plastic and Reconstructive Surgery, Kanagawa Children’s Medical Center, Yokohama, Kanagawa, Japan
#National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan

We recently developed a promising regenerative method based on the xenotransplantation of human cartilage progenitor cells, demonstrating self-renewing elastic cartilage reconstruction with expected long-term tissue restoration. However, it remains unclear whether autotransplantation of cartilage progenitors may work by a similar principle in immunocompetent individuals. We used a nonhuman primate (monkey) model to assess the safety and efficacy of our regenerative approach because the model shares characteristics with humans in terms of biological functions, including anatomical features. First, we identified the expandable and multipotent progenitor population from monkey ear perichondrium and succeeded in inducing chondrocyte differentiation in vitro. Second, in vivo transplanted progenitor cells were capable of reconstructing elastic cartilage by xenotransplantation into an immunodeficient mouse. Finally, the autologous monkey progenitor cells were transplanted into the subcutaneous region of a craniofacial section and developed mature elastic cartilage of their own 3 months after transplantation. Furthermore, we attempted to develop a clinically relevant, noninvasive monitoring method using magnetic resonance imaging (MRI). Collectively, this report shows that the autologous transplantation of cartilage progenitors is potentially effective for reconstructing elastic cartilage. This principle will be invaluable for repairing craniofacial injuries and abnormalities in the context of plastic and reconstructive surgery.

Key words: Autotransplantation; Monkey; Ear perichondrium-derived cartilage progenitor cells; Regenerative therapy; Plastic surgery

Received December 16, 2015; final acceptance February 7, 2016. Online prepub date: February 15, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Takanori Takebe, M.D., Associate Professor, Department of Regenerative Medicine Yokohama City University 3-9 Fuku-ura, Kanazawa-ku, Yokohama 236-0004, Japan. Tel: +81-45-787-2621; Fax: +81-45-787-8963; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Hideki Taniguchi, M.D., Ph.D. Professor, Department of Regenerative Medicine Yokohama City University 3-9 Fuku-ura, Kanazawa-ku, Yokohama 236-0004, Japan. Tel: +81-45-787-2621; Fax: +81-45-787-8963; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 963-971, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X688579
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Plasma Derived From Human Umbilical Cord Blood Modulates Mitogen-Induced Proliferation of Mononuclear Cells Isolated From the Peripheral Blood of ALS Patients

David J. Eve,* Jared Ehrhart,† Theresa Zesiewicz,‡ Israt Jahan,‡ Nicole Kuzmin-Nichols,† Cyndy Davis Sanberg,† Clifton Gooch,‡ Paul R. Sanberg,*§¶# and Svitlana Garbuzova-Davis*§¶

*Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, USA
Saneron CCEL Therapeutics, Inc., Tampa, FL, USA
‡Department of Neurology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA
§Department of Molecular Pharmacology and Physiology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA
¶Department of Pathology and Cell Biology, University of South Florida, Morsani College of Medicine, Tampa, FL, USA
#Department of Psychiatry, University of South Florida, Morsani College of Medicine, Tampa, FL, USA

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by degeneration of motor neurons in the spinal cord and brain. This disease clinically manifests as gradual muscular weakness and atrophy leading to paralysis and death by respiratory failure. While multiple interdependent factors may contribute to the pathogenesis of ALS, increasing evidence shows the possible presence of autoimmune mechanisms that promote disease progression. The potential use of plasma derived from human umbilical cord blood (hUCB) as a therapeutic tool is currently in its infancy. The hUCB plasma is rich in cytokines and growth factors that are required for growth and survival of cells during hematopoiesis. In this study, we investigated the effects of hUCB plasma on the mitogen-induced proliferation of mononuclear cells (MNCs) isolated from the peripheral blood of ALS patients and apoptotic activity by detection of caspase 3/7 expression of the isolated MNCs in vitro. Three distinct responses to phytohemagglutinin (PHA)-induced proliferation of MNCs were observed, which were independent of age, disease duration, and the ALS rating scale: Group I responded normally to PHA, Group II showed no response to PHA, while Group III showed a hyperactive response to PHA. hUCB plasma attenuated the hyperactive response (Group III) and potentiated the normal response in Group I ALS patients, but did not alter that of the nonresponders to PHA (Group II). The elevated activity of caspase 3/7 observed in the MNCs from ALS patients was significantly reduced by hUCB plasma treatment. Thus, study results showing different cell responses to mitogen suggest alteration in lymphocyte functionality in ALS patients that may be a sign of immune deficiency in the nonresponders and autoimmunity alterations in the hyperactive responders. The ability of hUCBplasma to modulate the mitogen cell response and reduce caspase activity suggests that the use of hUCB plasma alone, or with stem cells, may prove useful as a therapeutic in ALS patients.

Key words: Human umbilical cord blood plasma (hUCBP); Autoimmunity; PhytohemagglutininNonresponders; Caspase

Received May 24, 2015; final acceptance July 7, 2015. Online prepub date: July 8, 2015.
Address correspondence to Svitlana Garbuzova-Davis, Ph.D., D.Sc., Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA. Tel: +1-813-974-3189; Fax: +1-813-974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 973-982, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690520
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

miR-sc3, a Novel MicroRNA, Promotes Schwann Cell Proliferation and Migration by Targeting Astn1

Sheng Yi,*1 Shanshan Wang,†1 Qing Zhao,‡ Chun Yao,* Yun Gu,* Jie Liu,* Xiaosong Gu,* and Shiying Li*

*Jiangsu Key Laboratory of Neuroregeneration, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu, China
†Department of Obstetrics and Gynecology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China
‡Key Laboratory of the People’s Liberation Army, Institute of Orthopaedics, Chinese PLA General Hospital, Beijing, China

MicroRNAs (miRNAs, miRs) are small noncoding RNAs that regulate gene expression at the posttranscriptional level. We have previously identified a group of novel miRNAs in proximal sciatic nerve after sciatic nerve transection by Solexa sequencing, and miR-sc3 is a member of the group. In this study, we aimed to investigate the effects of miR-sc3 on phenotypic modulation of Schwann cells (SCs). miR-sc3 was highly expressed in the injured nerve after sciatic nerve transection. An increased and decreased expression of miR-sc3 promoted and reduced the proliferation and migration of primary SCs, respectively. miR-sc3 directly targeted astrotactin 1 (Astn1) and led to translational suppression of Astn1. There was an inverse association between the time-dependent expressions of miR-sc3 and Astn1 in proximal sciatic nerve after sciatic nerve transection. Overall, miR-sc3 affected SC proliferation and migration by targeting Astn1, thus playing the regulatory role in peripheral nerve regeneration.

Key words: miR-sc3; Schwann cells (SCs); Proliferation; Migration; Astrotactin 1 (Astn1)

Received December 16, 2015; final acceptance January 26, 2016. Online prepub date: January 18, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Prof. Xiaosong Gu, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, 19 Qixiu Road, Nantong, Jiangsu, China. Tel: +086-513-85051801; Fax: +086-513-85511585; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Shiying Li, Jiangsu Key Laboratory of Neuroregeneration, Nantong University, 19 Qixiu Road, Nantong, Jiangsu, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 983-993, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690494
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Repair of Rat Sciatic Nerve Defects by Using Allogeneic Bone Marrow Mononuclear Cells Combined With Chitosan/Silk Fibroin Scaffold

Min Yao,*1 Yi Zhou,*1 Chengbin Xue,* Hechun Ren,* Shengran Wang,* Hui Zhu,*† Xingjian Gu,* Xiaosong Gu,* and Jianhui Gu

*Jiangsu Key Laboratory of Neuroregeneration, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, China
†Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong, China
‡Department of Hand Surgery, Affiliated Hospital of Nantong University, Nantong, China

The therapeutic benefits of bone marrow mononuclear cells (BM-MNCs) in many diseases have been well established. To advance BM-MNC-based cell therapy into the clinic for peripheral nerve repair, in this study we developed a new design of tissue-engineered nerve grafts (TENGs), which consist of a chitosan/fibroin-based nerve scaffold and BM-MNCs serving as support cells. These TENGs were used for interpositional nerve grafting to bridge a 10-mm-long sciatic nerve defect in rats. Histological and functional assessments after nerve grafting showed that regenerative outcomes achieved by our developed TENGs were better than those achieved by chitosan/silk fibroin scaffolds and were close to those achieved by autologous nerve grafts. In addition, we used green fluorescent protein-labeled BM-MNCs to track the cell location within the chitosan/fibroin-based nerve scaffold and trace the cell fate at an early stage of sciatic nerve regeneration. The result suggested that BM-MNCs could survive at least 2 weeks after nerve grafting, thus helping to gain a preliminary mechanistic insight into the favorable effects of BM-MNCs on axonal regrowth.

Key words: Bone marrow mononuclear cells (BM-MNCs); Chitosan/silk fibroin scaffold; Rat sciatic nerve; Peripheral nerve repair

Received December 16, 2015; final acceptance January 26, 2016. Online prepub date: January 15, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Xiaosong Gu, Jiangsu Key Laboratory of Neuroregeneration, Co-innovation Center of Neuroregeneration, Nantong University, 19 Qixiu Road, Nantong, JS 226001, China. Tel: +86-513-85051801; Fax: +86-513-85511585; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Jianhui Gu, Department of Hand Surgery, Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, JS 226001, China. Tel: +86-513-85052066; Fax: +86-513-85519820; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it