Cell Transplantation 24(10) Abstracts

Return to Cell Transplantation>

Cell Transplantation, Vol. 24, pp. 1915-1930, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685744
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Stem Cell Therapy for Corneal Regeneration Medicine and Contemporary Nanomedicine for Corneal Disorders

Chih-Chien Hsu,*†1 Chi-Hsien Peng,†‡1 Kuo-Hsuan Hung,†§¶1 Yi-Yen Lee,†# Tai-Chi Lin,*† Shih-Fan Jang,#** Jorn-Hon Liu,†† Yan-Ting Chen,†‡‡§§ Lin-Chung Woung,¶¶ Chien-Ying Wang,¶#** Ching-Yao Tsa,¶¶ Shih-Hwa Chiou,*¶#** Shih-Jen Chen,*¶ and Yuh-Lih Chang¶#**

*Department of Ophthalmology, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
†Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan, ROC
‡Department of Ophthalmology, Shin Kong Wu Ho-Su Memorial Hospital and Fu-Jen Catholic University, Taipei, Taiwan, ROC
§Department of Ophthalmology, National Yang-Ming University Hospital, Yilan, Taiwan, ROC
¶School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC
#Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan, ROC
**Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan, ROC
††Department of Ophthalmology, Cheng-Hsin Hospital, Taipei, Taiwan, ROC
‡‡Department of Ophthalmology, Changhua Christian Hospital, Changhua, Taiwan, ROC
§§Department of Optometrics, Central Taiwan University of Science and Technology, Changhua, Taiwan, ROC
¶¶Department of Ophthalmology, Taipei City Hospital, Taipei, Taiwan, ROC

The ocular surface is the outermost part of the visual system that faces many extrinsic or intrinsic threats, such as chemical burn, infectious pathogens, thermal injury, Stevens–Johnson syndrome, ocular pemphegoid, and other autoimmune diseases. The cornea plays an important role in conducting light into the eyes and protecting intraocular structures. Several ocular surface diseases will lead to the neovascularization or conjunctivalization of corneal epithelium, leaving opacified optical media. It is believed that some corneal limbal cells may present stem cell-like properties and are capable of regenerating corneal epithelium. Therefore, cultivation of limbal cells and reconstruction of the ocular surface with these limbal cell grafts have attracted tremendous interest in the past few years. Currently, stem cells are found to potentiate regenerative medicine by their capability of differentiation into multiple lineage cells. Among these, the most common cell sources for clinical use are embryonic, adult, and induced stem cells. Different stem cells have varied specific advantages and limitations for in vivo and in vitro expansion. Other than ocular surface diseases, culture and transplantation of corneal endothelial cells is another major issue for corneal decompensation and awaits further studies to find out comprehensive solutions dealing with nonregenerative corneal endothelium. Recently, studies of in vitro endothelium culture and r-associated kinase (ROCK) inhibitor have gained encouraging results. Some clinical trials have already been finished and achieved remarkable vision recovery. Finally, nanotechnology has shown great improvement in ocular drug delivery systems during the past two decades. Strategies to reconstruct the ocular surface could combine with nanoparticles to facilitate wound healing, drug delivery, and even neovascularization inhibition. In this review article, we summarized the major advances of corneal limbal stem cells, limbal stem cell deficiency, corneal endothelial cell culture/transplantation, and application of nanotechnology on ocular surface reconstruction. We also illustrated potential applications of current knowledge for the future treatment of ocular surface diseases.

Key words: Corneal neovascularization; Stem cells; Nanomedicine; Regeneration medicine; Corneal endothelial cell transplant

Received July 10, 2014; final acceptance November 25, 2014. Online prepub date: December 12, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Yuh-Lih Chang, Ph.D., Institute of Pharmacology, School of Medicine, National Yang-Ming University, No. 155, Sec. 2, Linong Street, Taipei, 11221 Taiwan, ROC. Tel: +886-2-2875-7325; Fax: +886-2-2876-1351; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1931-1943, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685410
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Ex Vivo Expanded Allogeneic Mesenchymal Stem Cells With Bone Marrow Transplantation Improved Osteogenesis in Infants With SevereHypophosphatasia

Takeshi Taketani,*† Chigusa Oyama,† Aya Mihara,† Yuka Tanabe,† Mariko Abe,† Tomohiro Hirade,† Satoshi Yamamoto,† Ryosuke Bo,† Rie Kanai,† Taku Tadenuma,‡ Yuko Michibata,‡ SoichiroYamamoto,§ Miho Hattori,* Yoshihiro Katsube,¶ Hiroe Ohnishi,¶ Mari Sasao,¶ Yasuaki Oda,¶ Koji Hattori,¶ Shunsuke Yuba,¶ Hajime Ohgushi,¶ and Seiji Yamaguchi†

*Division of Blood Transfusion, Shimane University Hospital, Izumo, Shimane, Japan
†Department of Pediatrics, Shimane University School of Medicine, Izumo, Shimane, Japan
‡Division of Rehabilitation, Shimane University Hospital, Izumo, Shimane, Japan
§Department of Orthopedics, Shimane University School of Medicine, Izumo, Shimane, Japan
¶Department of Life Science and Biotechnology, National Institute of Advanced Industrial Science and Technology, Amagasaki, Hyogo, Japan

Patients with severe hypophosphatasia (HPP) develop osteogenic impairment with extremely low alkaline phosphatase (ALP) activity, resulting in a fatal course during infancy. Mesenchymal stem cells (MSCs) differentiate into various mesenchymal lineages, including bone and cartilage. The efficacy of allogeneic hematopoietic stem cell transplantation for congenital skeletal and storage disorders is limited, and therefore we focused on MSCs for the treatment of HPP. To determine the effect of MSCs on osteogenesis, we performed multiple infusions of ex vivo expanded allogeneic MSCs for two patients with severe HPP who had undergone bone marrow transplantation (BMT) from asymptomatic relatives harboring the heterozygous mutation. There were improvements in not only bone mineralization but also muscle mass, respiratory function, and mental development, resulting in the patients being alive at the age of 3. After the infusion of MSCs, chimerism analysis of the mesenchymal cell fraction isolated from bone marrow in the patients demonstrated that donor-derived DNA sequences existed. Adverse events of BMT were tolerated, whereas those of MSC infusion did not occur. However, restoration of ALP activity was limited, and normal bony architecture could not be achieved. Our data suggest that multiple MSC infusions, following BMT, were effective and brought about clinical benefits for patients with lethal HPP. Allogeneic MSC-based therapy would be useful for patients with other congenital bone diseases and tissue disorders if the curative strategy to restore clinically normal features, including bony architecture, can be established.

Key words: Hypophosphatasia (HPP); Bone marrow transplantation (BMT); Mesenchymal stem cell (MSC) transplantation; Osteogenesis

Received July 12, 2014; final acceptance November 5, 2014. Online prepub date: November 13, 2014.
Address correspondence to Takeshi Taketani, M.D., Ph.D., Division of Blood Transfusion, Shimane University Hospital, 89-1, Enya, Izumo, Shimane 693-8501, Japan. Tel: +81-853-20-2409; Fax: +81-853-20-2409; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1945-1955, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685276
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bone-Forming Capacity and Biodistribution of Bone Marrow-Derived Stromal Cells Directly Loaded Into Scaffolds: A Novel and Easy Approach for Clinical Application of Bone Regeneration

Julie Léotot,*† Angélique Lebouvier,*† Philippe Hernigou,*‡ Philippe Bierling,†§ Hélène Rouard,*†¶ and Nathalie Chevallier*†

*Université Paris-Est Créteil, Faculté de médecine, Laboratoire de “Bioingénierie Cellulaire, Tissulaire et Sanguine,” Créteil, France
†Etablissement Français du Sang d’Ile-de-France, Unité d’Ingénierie et de Thérapie Cellulaire, Créteil, France
‡Service de Chirurgie Orthopédique et Traumatologique, AP-HP Hôpital Henri-Mondor, Créteil, France
§INSERM UMR955, Paris-Est University, Créteil, France
¶AP-HP Hôpital Henri-Mondor – A. Chenevier, Service Hospitalier, Créteil, France

In the context of clinical applications of bone regeneration, cell seeding into scaffolds needs to be safe and easy. Moreover, cell density also plays a crucial role in the development of efficient bone tissue engineering constructs. The aim of this study was to develop and evaluate a simple and rapid cell seeding procedure on hydroxyapatite/β-tricalcium phosphate (HA/βTCP), as well as define optimal cell density and control the biodistribution of grafted cells. To this end, human bone marrow-derived stromal cells (hBMSCs) were seeded on HA/βTCP scaffolds, and we have compared bone formation using an ectopic model. Our results demonstrated a significantly higher bone-forming capacity of hBMSCs directly loaded on HA/βTCP during surgery compared tohBMSCs preseeded for 7 days in vitro on HA/βTCP before ectopic implantation. The extent of new bone formation increases with increasing hBMSC densities quantitatively, qualitatively, and in frequency. Also, this study showed that grafted hBMSCsremained confined to the implantation site and did not spread toward other tissues, such as liver, spleen, lungs, heart, and kidneys. In conclusion, direct cell loading into a scaffold during surgery is more efficient for bone regeneration, as well as quick and safe. Therefore direct cell loading is suitable for clinical requirements and cell production control, making it a promising approach for orthopedic applications. Moreover, our results have provided evidence that the formation of a mature bone organ containing hematopoietic islets needs a sufficiently high local density of grafted hBMSCs, which should guide the optimal dose of cells for clinical use.

Key words: Bone marrow-derived stromal cell transplantation; Cell seeding; Cell density; Biodistribution; Bone tissue engineering

Received May 7, 2014; final acceptance October 21, 2014. Online prepub date: October 28, 2014.
Address correspondence to Nathalie Chevallier, Etablissement Français du Sang d’Ile-de-France, Unité d’Ingénierie et de Thérapie Cellulaire, 5 rue Gustave Eiffel, 94017 Créteil cedex, France. Tel: +33 1 56 72 21 20; Fax: +33 1 56 72 76 88; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1957-1968, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685393
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Dose–Effect Relationship Between the Seeding Quantity of Human Marrow Mesenchymal Stem Cells and In Vivo Tissue-Engineered Bone Yield

Huanhuan Wu, Ning Kang, Qian Wang, Ping Dong, Xiaoyan Lv, Yilin Cao, and Ran Xiao

Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P. R. China

Although the feasibility of human bone marrow mesenchymal stem cell (hBMMSC)-based tissue-engineered bone (TEB) has been proven in a number of studies, reaching a high positive fraction and bone yield of TEB still remains a challenge. Here we report a dose–effect relationship of the quantity of seeded cells with in vivo bone yield and the required quantity of hBMMSCs for the effective, stable bone formation of TEB. In our study, TEB was constructed using the static seeding technique with the gradient of seeding densities and volumes of passage 3 hBMMSCs. The in vitro characteristics of seeding efficiency, proliferation, viability, distribution, and osteogenic differentiation of hBMMSCs seeded on two commercial scaffolds of β-TCP and CHA were investigated using alamarBlue assay, live/dead staining, confocal laser scanning microscope, scanning electronic microscopy examination, and mRNA expression analysis of osteogenic differentiation markers. After 3 months of ectopic implantation, in vivo bone regeneration was examined by quantitative analysis of histology and micro-CT. The results showed that 10 × 106 cells/ml was the minimum cell seeding density for CHA and β-TCP to generate new bone in vivo. In addition, 20 × 106 cells/ml and 30 × 106cells/ml were the saturating seeding densities for CHA and β-TCP to produce new bone effectively and stably, respectively. Thus, for different scaffolds, the saturating seeding density should be investigated first to ensure the effectiveness and stability of TEB construction with minimum donor injury, which is essential for the clinical application of TEB.

Key words: Bone tissue engineering; Bone marrow mesenchymal stem cells; Ectopic bone yield; Dose–effect relationship

Received May 12, 2014; final acceptance November 4, 2014. Online prepub date: November 13, 2014.
Address correspondence to Yilin Cao, Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100144, P. R. China. Tel: +86 10 88771507; Fax: +86 10 88960373; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Ran Xiao, Research Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100144, P. R. China. Tel: +86 10 88771507; Fax: +86 10 88960373; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1969-1980, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685294
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Canine Mesenchymal Stem Cell Potential and the Importance of Dog Breed: Implication for Cell-Based Therapies

Alessandro Bertolo,* Frank Steffen,† Cherry Malonzo-Marty,* and Jivko Stoyanov*‡

*Swiss Paraplegic Research, Nottwil, Switzerland
†Vetsuisse faculty of the University of Zurich, Zurich, Switzerland
‡Institute for Surgical Technology and Biomechanics, University of Bern, Bern, Switzerland

The study of canine bone marrow-derived mesenchymal stem cells (MSCs) has a prominent position in veterinary cell-based applications. Yet the plethora of breeds, their different life spans, and interbreed variations provide unclearness on what can be achieved specifically by such therapies. In this study, we compared a set of morphological, physiological, and genetic markers of MSCs derived from large dog breeds, namely, Border collie, German shepherd, Labrador, Malinois, Golden retriever, andHovawart. We compared colony-forming units (CFUs) assay, population doubling time (PDT), senescence-associated β-galactosidase (SA-β-gal) activity, telomere length, and gene expression of MSCs, as well as the ability of cells to differentiate to osteogenic, adipogenic, and chondrogenic phenotypes. The influence of the culture media a-MEM, low-glucose DMEM, and high-glucose DMEM, used in cell isolation and expansion, was investigated in the presence and absence of basic fibroblast growth factor (bFGF). Initial cell yield was not affected by culturing medium, but MSCs expanded best in a-MEM supplemented with bFGF. After isolation, the number of MSCs was similar among breeds—as shown by equivalent CFUs—except in the Hovawartsamples, which had fivefold less CFU. Telomere lengths were similar among breeds. MSCs divided actively only for 4 weeks in culture (PDT = ~50 h/division), except Border collie cells divided for a longer time than cells from other groups. The percentage of senescent cells increased linearly in all breeds with time, with a faster rate in German shepherd, Labrador, and Golden retriever. Border collie cells underwent efficient osteogenic differentiation, Hovawart cells performed the best in chondrogenicdifferentiation, and Labrador cells in both, while German shepherd cells had the lower differentiation potential. MSCs from all breeds preserved the same adipogenic differentiation potential. In conclusion, despite variations, isolated MSCs can be expanded and differentiated in vitro, and all breeds are eligible for MSC-based therapies.

Key words: Canine bone marrow stem cells; Chondrogenesis; Osteogenesis; Adipogenesis; Dog breeds

Received May 26, 2014; final acceptance October 28, 2014. Online prepub date: November 5, 2014.
Address correspondence to Jivko Stoyanov, Biomedical Laboratories, Swiss Paraplegic Research, G. A. Zäch Strasse 4, CH-6207 Nottwil, Switzerland. Tel: +41 41-939-6635; Fax: +41 41-939-6640; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1981-1997, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685302
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of Magnetic Intensities for Mesenchymal Stem Cell Targeting Therapy on Ischemic Myocardial Repair: High Magnetic Intensity Improves Cell Retention but Has no Additional Functional Benefit

Yunli Shen,*1 Xuebo Liu,*1 Zheyong Huang,† Ning Pei,‡ Jianfeng Xu,† Zheng Li,§ Yunkai Wang,* Juying Qian,† and Junbo Ge†

*Department of Cardiology, Shanghai East Hospital, Tongji University, Shanghai, China
†Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China
‡College of Science, Shanghai University, Shanghai, China
§Biomedical Research Center, Zhongshan Hospital, Fudan University, Shanghai, China

Magnetic targeting has the potential to enhance the therapeutic effects of stem cells through increasing retention of transplanted cells. To investigate the effects of magnetic targeting intensities on cell transplantation, we performed different magnetic intensities for mesenchymal stem cell (MSC)-targeting therapy in a rat model of ischemia/reperfusion. Rat MSCs labeled with superparamagnetic oxide nanoparticles (SPIOs) were injected into the left ventricular (LV) cavity of rats during a brief aorta and pulmonary artery occlusion. The 0.15 Tesla (T), 0.3 T, and 0.6 T magnets were placed 0~1 mm above the injured myocardium during and after the injection of 1 × 106 MSCs. Fluorescence imaging and quantitative PCR revealed that magnetic targeting enhanced cell retention in the heart at 24 h in a magnetic field strength-dependent manner. Compared with the 0 T group, three magnetic targeting groups enhanced varying cell engraftment at 3 weeks, at which time LV remodeling was maximally attenuated, and the therapeutic benefit (LV ejection fraction) was also highest in the 0.3 T groups. Interestingly, due to the low MSC engraftment resulting from microvascular embolisms, the 0.6 T group failed to translate into additional therapeutic outcomes, though it had the highest cell retention. Magnetic targeting enhances cell retention in a magnetic field strength-dependent manner. However, too high of a magnetic intensity may result in microembolization and consequently undermine the functional benefits of cell transplantation.

Key words: Mesenchymal stem cells; Magnetic targeting intensity; Microembolization; Myocardial infarction; Rat model

Received July 24, 2013; final acceptance September 20, 2014. Online prepub date: November 5, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Juying Qian, M.D., Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 180 Feng Lin Road, Shanghai 200032, China. Tel: +86-21-64041990, x10564; Fax: +86-21-64223006; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or ZheyongHuang, M.D., Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 180 Feng Lin Road, Shanghai 200032, China. Tel: +86-21-64041990, x12596; Fax: +86-21-64223006; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 1999-2010, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685267
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Mesenchymal Stem Cells Enhance Angiogenesis and Follicle Survival in Human Cryopreserved Ovarian Cortex Transplantation

Xi Xia,*†‡ Tailang Yin,*§ Jie Yan,*§ Liying Yan,*§ Chao Jin,¶ Cuilin Lu,*§ Tianren Wang,*§ Xiaohui Zhu,*‡§ Xu Zhi,*‡§ Jijun Wang,*§# Lei Tian,*§# Jing Liu,¶ Rong Li,*‡§ and Jie Qiao*‡§

*Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, China
†Department of Reproductive Medicine, Peking University Shenzhen Hospital, Shenzhen, China
‡Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, China
§Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, Beijing, China
¶Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing, China
#Department of Hematology, Peking University Third Hospital, HaiDian District, Beijing, China

Transplantation of cryopreserved ovarian tissue is a novel technique to restore endocrine function and fertility especially for cancer patients. However, the main obstacle of the technique is massive follicle loss as a result of ischemia in the process of transplantation. Mesenchymal stem cells (MSCs) have been acknowledged to play an important role in supporting angiogenesis and stabilizing long-lasting blood vessel networks through release of angiogenic factors and differentiation into pericytes and endothelial cells. This study is aimed to investigate whether MSCs could be applied to overcome the above obstacle to support the ovarian tissue survival in the transplantation. Here we show that human MSCs could enhance the expression level of VEGF, FGF2, and especially the level of angiogenin, significantly stimulate neovascularization, and increase blood perfusion of the grafts in the cryopreserved ovarian tissue transplantation. Further studies reveal that MSCs could notably reduce the apoptotic rates of primordial follicles and decrease follicle loss in the grafted ovarian tissues. In summary, our findings demonstrate a previously unrecognized function of MSCs in improving human ovarian tissue transplantation and provide a useful strategy to optimize fertility preservation and restoration.

Key words: Ovarian cortex transplantation; Mesenchymal stem cells (MSCs); Fertility preservation; Ovarian cryopreservation; Follicle survival

Received June 26, 2014; final acceptance October 15, 2014. Online prepub date: October 28, 2014.
Address correspondence to Jie Qiao, Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, No. 49 North HuaYuan Road, HaiDian District, Beijing 100191, China. Fax: +86-10-82265080; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Xi Xia, Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, No. 49 North HuaYuan Road, HaiDian District, Beijing 100191, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2011-2028, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685258
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

IL-1β-Induced Mesenchymal Stem Cell Migration Involves MLCK Activation via PKC Signaling

Cheng-Yu Lin,* Chia-Hua Zu,* Chih-Chang Yang,* Pei-Jiun Tsai,† Jia-Fwu Shyu,‡ Chie-Pein Chen,§ Zen-Chung Weng,¶ Tien-Hua Chen,*# and Hwai-Shi Wang*

*Institute of Anatomy and Cell Biology, School of Medicine, National Yang Ming University, Taipei, Taiwan
†Institute of Clinical Medicine, National Yang Ming University, Taipei, Taiwan
‡Department of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan
§Division of High Risk Pregnancy, Mackay Memorial Hospital, Taipei, Taiwan
¶Division of Cardiovascular Surgery, Cardiovascular Center, Taipei Medical University Hospital, Taipei, Taiwan
#Department of Surgery, Veteran General Hospital, Taipei, Taiwan

Mesenchymal stem cells (MSCs) migrate via the bloodstream to sites of injury, possibly attracted by inflammatory cytokines. Although many cytokines can induce stem cell migration, the underlying mechanism is not fully understood. We found that tail vein-injected MSCs migrate to the pancreas in nonobese diabetic (NOD) mice. An ELISA assay revealed that hyperglycemic NOD mice have higher pancreatic levels of interleukin-1β (IL-1β) than normal NOD mice and that IL-1β stimulates MSC migration in aTranswell assay and electric cell–substrate impedance sensing system. Microarray analysis showed that myosin light chain kinase (MLCK) is involved in IL-1β-induced MSC migration, while Western blots showed that IL-1β stimulates MLCK expression and activation and that MLCK-siRNA transfection reduces MSC migration. Kinase inhibitors, chromatin immunoprecipitation, and a knockdown study revealed that IL-1β-induced MLCK expression is regulated by the PKCδ/NF-κB signaling pathway, and a kinase inhibitor study revealed that IL-1β-induced MLCK activation occurs via the PKCα/MEK/ERK signaling pathway. These results show that IL-1β released from the pancreas of hyperglycemic NOD mice induces MSC migration and that this is dependent on MLCK expression via the PKCδ/NF-κB pathway and on MLCK activation via the PKCα/MEK/ERK signaling cascade. This study increases our understanding of the mechanisms by which MSCs home to injury sites.

Key words: Interleukin-1β (IL-1β); Myosin light chain kinase (MLCK); Protein kinase C (PKC) signaling; Stem cell migration; Nonobese diabetic (NOD) mice

Received April 17, 2014; final acceptance October 12, 2014. Online prepub date: October 20, 2014.
Address correspondence to Hwai-Shi Wang, Ph.D., Department of Anatomy, School of Medicine, Yang Ming University, Peitou, Taipei, Taiwan 112, R.O.C. Tel: +(886) 2228267035; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Tien-Hua Chen, M.D., Department of Anatomy, National Yang Ming University, Taipei, Taiwan 112, R.O.C. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2029-2039, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685401
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Concentration-Dependent Vascularization of Adipose Stromal Vascular Fraction Cells

John G. Maijub,1 Nolan L. Boyd,1 Jacob R. Dale, James B. Hoying, Marvin E. Morris, and Stuart K. Williams

Cardiovascular Innovation Institute, Department of Surgery, University of Louisville School of Medicine, Louisville, KY, USA

Adipose-derived stromal vascular fraction (SVF) cells have been shown to self-associate to form vascular structures under both in vitro and in vivo conditions. The angiogenic (new vessels from existing vessels) and vasculogenic (new vessels through self-assembly) potential of the SVF cell population may provide a cell source for directly treating (i.e., point of care without further cell isolation) ischemic tissues. However the correct dosage of adipose SVF cells required to achieve a functional vasculature has not been established. Accordingly, in vitro and in vivo dose response assays were performed evaluating the SVF cell vasculogenic potential. Serial dilutions of freshly isolated rat adipose SVF cells were plated on growth factor reduced Matrigel andvasculogenesis, assessed as cellular tube-like network assembly, was quantified after 3 days of culture. This in vitro vasculogenesis assay indicated that rat SVF cells reached maximum network length at a concentration of 2.5 × 105 cells/ml and network maintained at the higher concentrations tested. The same concentrations of rat and human SVF cells were used to evaluate vasculogenesis in vivo. SVF cells were incorporated into collagen gels and subcutaneously implanted into Rag1 immunodeficient mice. The 3D confocal images of harvested constructs were evaluated to quantify dose dependency of SVF cell vasculogenesis potential. Rat- and humanderived SVF cells yielded a maximum vasculogenic potential at 1 × 106 and 4 × 106 cells/ml, respectively. No adverse reactions (e.g., toxicity, necrosis, tumor formation) were observed at any concentration tested. In conclusion, the vasculogenic potential of adipose-derived SVF cell populations is dose dependent.

Key words: Adipose stromal vascular fraction; Therapeutic angiogenesis; Vasculogenesis

Received January 24, 2014; final acceptance November 5, 2014. Online prepub date: November 13, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Stuart K. Williams, Ph.D., Cardiovascular Innovation Institute, 302 E. Muhammad Ali Blvd., Louisville, KY 40202, USA. Tel: +1-502-852-1379; Fax: +1-502-852-1795; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2041-2053, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685582
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Intrarenal Delivery of Mesenchymal Stem Cells and Endothelial Progenitor Cells Attenuates Hypertensive Cardiomyopathy in ExperimentalRenovascular Hypertension

Alfonso Eirin,* Xiang-Yang Zhu,* Behzad Ebrahimi,* James D. Krier,* Scott M. Riester,† Andre J. van Wijnen,† Amir Lerman,‡ and Lilach O. Lerman*‡

*Department of Internal Medicine, Division of Nephrology and Hypertension, Mayo Clinic, Rochester, MN, USA
†Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA
‡Division of Cardiovascular Diseases, Mayo Clinic, Rochester, MN, USA

Renovascular hypertension (RVH) leads to left ventricular (LV) hypertrophy and diastolic dysfunction, associated with increased cardiovascular mortality. Intrarenal delivery of endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) improves kidney function in porcine RVH, and the potent anti-inflammatory properties of MSCs may serve to blunt inflammatory mediators in the cardiorenal axis. However, their relative efficacy in attenuating cardiac injury and dysfunction remains unknown. This study tested the hypothesis that the cardioprotective effect of EPCs and MSCs delivered into the stenotic kidney in experimental RVH are comparable. Pigs (n = 7 per group) were studied after 10 weeks of RVH or control untreated or treated with a single intrarenal infusion of autologous EPCs or MSCs 4 weeks earlier. Cardiac and renal function (fast CT) and stenotic kidney release of inflammatory mediators (ELISA) were assessed in vivo, and myocardial inflammation, remodeling, and fibrosis ex vivo. After 10 weeks of RVH, blood pressure was not altered in cell-treated groups, yet stenotic kidney glomerular filtration rate (GFR), blunted in RVH, improved in RVH + EPC, and normalized in RVH + MSCs. Stenotic kidney release of monocyte chemoattractant protein (MCP)-1 and its myocardial expression were elevated in RVH + EPC, but normalized only in RVH + MSC pigs. RVH-induced LV hypertrophy was normalized in both EPC- and MSC-treated pigs, while diastolic function (E/A ratio) was restored to normal levels exclusively in RVH + MSCs. RVH-induced myocardial fibrosis and collagen deposition decreased in RVH + EPCs but further decreased in RVH + MSC-treated pigs. Intrarenal delivery of EPCs or MSCs attenuates RVH-induced myocardial injury, yet MSCs restore diastolic function more effectively than EPCs, possibly by greater improvement in renal function or reduction of MCP-1 release from the stenotic kidney. These observations suggest a therapeutic potential for EPCs and MSCs in preserving the myocardium in chronic experimental RVH.

Key words: Renal hypertension; Myocardium; Stem cells; Progenitor cells

Received February 14, 2014; final acceptance November 10, 2014. Online prepub date: November 21, 2014.
Address correspondence to Lilach O. Lerman, M.D., Ph.D., Division of Nephrology and Hypertension, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. E-mail: Lerman.Lilach@Mayo.


Cell Transplantation, Vol. 24, pp. 2055-2064, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685609
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Transplanted Human Amniotic Epithelial Cells Secrete Paracrine Proangiogenic Cytokines in Rat Model of Myocardial Infarction

Yi-Sun Song,* Hyun-Woo Joo,* In-Hwa Park,* Guang-Yin Shen,† Yonggu Lee,† Jeong Hun Shin,† Hyuck Kim,‡ Il-Seob Shin,§ and Kyung-Soo Kim*†

*Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, South Korea
†Cardiology Division, Department of Internal Medicine, Hanyang University College of Medicine, Seoul, South Korea
‡Department of Thoracic and Cardiovascular Surgery, Hanyang University College of Medicine, Seoul, South Korea
§Stem Cell Research Center, K-STEMCELL, Seoul, South Korea

Human amniotic epithelial cells (h-AECs) have been shown to differentiate into cardiomyocyte-like cells in vivo that can regenerate myocardial tissue and improve cardiac function in a rat model of myocardial infarction (MI). In this study, we investigated the paracrine factors released from h-AECs under hypoxic conditions to elucidate the possible mechanisms underlying this previously reported phenomenon of h-AEC-mediated cardiac repair. We used hypoxic cell culture conditions to simulate myocardial infarction in vitro. In comparison to normal conditions, we found that h-AECs secreted higher levels of several cytokines, including angiogenin (ANG), epidermal growth factor (EGF), interleukin (IL)-6, and monocyte chemoattractant protein (MCP)-1. To determine whether transplanted h-AECs express these proangiogenic cytokines in vivo, we ligated the coronary artery of rats to cause MI and injected either h-AECs or saline into the infarcted area. We found that the infarct and border zones of rat myocardium treated with h-AECs had higher expression levels of the human-origin cytokines ANG, EGF, IL-6, and MCP-1 compared to the tissues of saline-treated rats. In conclusion, h-AECs secreted proangiogenic cytokines in a rat model of MI, which may suggest that the paracrine effect by h-AECs could regenerate myocardial tissue and improve cardiac function.

Key words: Amniotic epithelial cells; Myocardial infarction (MI); Paracrine; Cytokine

Received May 27, 2014; final acceptance November 16, 2014. Online prepub date: November 21, 2014.
Address correspondence to Kyung-Soo Kim, Cardiology Division, Department of Internal Medicine, Hanyang University College of Medicine, 17 Haengdang-dong, Sungdong-ku, Seoul 133-792, South Korea. Tel: +82-2-2290-8312; Fax: +82-2-2298-9183; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2065-2076, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685230
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Amnion-Derived Stem Cells Have Immunosuppressive Properties on NK Cells and Monocytes

Jiali Li,* Chika Koike-Soko,* Jun Sugimoto,† Toshiko Yoshida,* Motonori Okabe,* and Toshio Nikaido*

*Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan
†Department of Human Molecular Biology, University of the Ryukyus, Okinawa, Japan

Human amnion-derived cells are considered to be a promising alternative cell source for their potential clinical use in tissue engineering and regenerative medicine because of their proliferation and differentiation ability. The cells can easily be obtained from human amnion, offering a potential source without medical intervention. It has been proven that human amnion-derived cells express immunosuppressive factors CD59 and HLA-G, implying that they may have an immunosuppressive function. To assess the immunosuppressive activity, we investigated the effect of human amnion-derived cells on NK cell and monocyte function. Amnion-derived cells inhibited the cytotoxicity of NK cells to K562 cells. The inhibition depended on the NK/amnion-derived cell ratio. The inhibition of NK cytotoxicity was recovered by continuous culturing without amnion-derived cells. The inhibition of NK cytotoxicity was related to the downregulation of the expression of the activated NK receptors and the production of IFN-γ, as well as the upregulation of the expression of IL-10 and PGE2 in human amnion-derived cells. The addition of antibody to IL-10 or PGE2 inhibitor tended to increase NK cytotoxicity. IL-10 and PGE2 might be involved in the immunosuppressive activity of amniotic cells toward NK cells. Amniotic cells also suppressed the activity of cytokine production in monocytes analyzed with TNF-α and IL-6. These data suggested that amniotic cells have immunosuppressive activity.

Key words: Amnion mesenchymal stem cells; Amnion epithelial stem cells; Immunosuppressive activity; IL-10; PGE2

Received April 22, 2014; final acceptance October 10, 2014. Online prepub date: October 20, 2014.
Address correspondence to Toshio Nikaido, Ph.D., Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan. Tel: +81-76-434-7210; Fax: +81-76-434-5011; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2077-2098, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685104
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Wharton’s Jelly-Derived Stem Cells Display Immunomodulatory Properties and Transiently Improve Rat Experimental Autoimmune Encephalomyelitis

Raf Donders,* Marjan Vanheusden,* Jeroen F. J. Bogie,* Stylianos Ravanidis,* Kristof Thewissen,* Piet Stinissen,* Wilfried Gyselaers,*† Jerome J. A. Hendriks,* and Niels Hellings*

*Hasselt University, Biomedical Research Institute/Transnational University Limburg, School of Life Sciences, Diepenbeek, Belgium
†Ziekenhuis Oost-Limburg, Campus St. Jan, Genk, Belgium

Umbilical cord matrix or Wharton’s jelly-derived stromal cells (WJ-MSCs) are an easily accessible source of mesenchymal-like stem cells. Recent studies describe a hypoimmunogenic phenotype, multipotent differentiation potential, and trophic support function for WJ-MSCs, with variable clinical benefit in degenerative disease models such as stroke, myocardial infarction, and Parkinson’s disease. It remains unclear whether WJ-MSCs have therapeutic value for multiple sclerosis (MS), where autoimmune-mediated demyelination and neurodegeneration need to be halted. In this study, we investigated whether WJ-MSCs possess the required properties to effectively and durably reverse these pathological hallmarks and whether they survive in an inflammatory environment after transplantation. WJ-MSCs displayed a lowly immunogenic phenotype and showed intrinsic expression of neurotrophic factors and a variety of anti-inflammatory molecules. Furthermore, they dose-dependently suppressed proliferation of activated T cells using contact-dependent and paracrine mechanisms. Indoleamine 2,3-dioxygenase 1 was identified as one of the main effector molecules responsible for the observed T-cell suppression. The immune-modulatory phenotype of WJ-MSCs was further enhanced after proinflammatory cytokine treatment in vitro (licensing). In addition to their effect on adaptive immunity, WJ-MSCs interfered with dendritic cell differentiation and maturation, thus directly affecting antigen presentation and therefore T-cell priming. Systemically infused WJ-MSCs potently but transiently ameliorated experimental autoimmune encephalomyelitis (EAE), an animal model for MS, when injected at onset or during chronic disease. This protective effect was paralleled with a reduction in autoantigen-induced T-cell proliferation, confirming their immunomodulatory activity in vivo. Surprisingly, in vitro licensed WJ-MSCs did not ameliorate EAE, indicative of a fast rejection as a result of enhanced immunogenicity. Collectively, we show that WJ-MSCs have trophic support properties and effectively modulate immune cell functioning both in vitro and in the EAE model, suggesting WJ-MSC may hold promise for MS therapy. Future research is needed to optimize survival of stem cells and enhance clinical durability.

Key words: Multiple sclerosis (MS); Wharton’s jelly; Transplantation; Immunomodulation; Mesenchymal stem cells (MSCs); Cell therapy

Received April 16, 2014; final acceptance September 26, 2014. Online prepub date: October 10, 2014.
Address correspondence to Prof. Dr. Niels Hellings, Hasselt University, Biomedical Research Institute, Campus Diepenbeek, Agoralaan building C, 3590 Diepenbeek, Belgium. Tel: +32 11 26 92 01; Fax: +32 11 26 92 99; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2099-2098, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685591
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Striatal Transplantation of Human Dopaminergic Neurons Differentiated From Induced Pluripotent Stem Cells Derived From Umbilical Cord Blood Using Lentiviral Reprogramming

Anna Effenberg,*1 Nancy Stanslowsky,†‡1 Alexander Klein,* Maike Wesemann,* Alexandra Haase,§¶ Ulrich Martin,§¶ Reinhard Dengler,†‡ Claudia Grothe,*‡ Andreas Ratzka,*2 and Florian Wegner†‡2

*Institute of Neuroanatomy, Hannover Medical School, Hannover, Germany
†Department of Neurology, Hannover Medical School, Hannover, Germany
‡Center for Systems Neuroscience (ZSN), Hannover Medical School, Hannover, Germany
§Department of Cardiac, Thoracic, Transplantation, and Vascular Surgery, Leibnitz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Hannover Medical School, Hannover, Germany
¶REBIRTH-Cluster of Excellence, Hannover, Germany

Human induced pluripotent stem cells (hiPSCs) are promising sources for regenerative therapies like the replacement of dopaminergic neurons in Parkinson’s disease. They offer an unlimited cell source that can be standardized and optimized to produce applicable cell populations to gain maximal functional recovery. In the present study, human cord blood-derived iPSCs (hCBiPSCs) were differentiated into dopaminergic neurons utilizing two different in vitro protocols for neural induction: (protocol I) by fibroblast growth factor (FGF-2) signaling, (protocol II) by bone morphogenetic protein (BMP)/transforming growth factor (TGF-β) inhibition. After maturation, in vitro increased numbers of tyrosine hydroxylase (TH)-positive neurons (7.4% of total cells) were observed by protocol II compared to 3.5% in protocol I. Furthermore, 3 weeks after transplantation in hemiparkinsonian rats in vivo, a reduced number of undifferentiated proliferating cells was achieved with protocol II. In contrast, proliferation still occurred in protocol I-derived grafts, resulting in tumor-like growth in two out of four animals 3 weeks after transplantation. Protocol II, however, did not increase the number of TH+ cells in the striatal grafts of hemiparkinsonian rats. In conclusion, BMP/TGF-β inhibition was more effective than FGF-2 signaling with regard to dopaminergic induction of hCBiPSCs in vitro and prevented graft overgrowth in vivo.

Key words: Dopaminergic neurons; Human cord blood cells; Intrastriatal transplantation; In vitro differentiation; Induced pluripotent stem cells (iPSCs); Parkinson’s disease

Received April 30, 2014; final acceptance November 7, 2014. Online prepub date: November 21, 2014.
1These authors provided equal contribution to this work.
2These senior authors provided equal contribution to this work.

Address correspondence to Dr. Andreas Ratzka, Institute of Neuroanatomy, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Tel: ++49/511/5322894; Fax: ++49/511/5322880; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2113-2128, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683584
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Dopamine Release Impairment in Striatum After Different Levels of Cerebral Cortical Fluid Percussion Injury

Yuan-Hao Chen,* Eagle Yi-Kung Huang,† Tung-Tai Kuo,‡ Hsin-I Ma,* Barry J. Hoffer,§ Pi-Fen Tsui,† Jing-Jr Tsai,* Yu-Ching Chou,¶ and Yung-Hsiao Chiang#

*Department of Neurological Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, R.O.C.
†Department of Pharmacology, National Defense Medical Center, Taipei, Taiwan, R.O.C.
‡Graduate Institute of Computer and Communication Engineering, National Taipei University of Technology, Taipei, Taiwan, R.O.C.
§Department of Neurosurgery, Case Western Reserve University School of Medicine, Cleveland, OH, USA
¶School of Public Health, National Defense Medical Center, Taipei, Taiwan, R.O.C.
#Graduate Program on Neuroregeneration, Taipei Medical University, Taipei, Taiwan, R.O.C.

To investigate the role of dopamine release in cognitive impairment and motor learning deficits after brain injury, different levels of traumatic brain injury (TBI) were made in rats by using fluid percussion at two different atmospheres (2 Psi and 6 Psi). Tonic and phasic bursting dopamine release and behavior tests followed at several time points. We used in vitro fast-scan cyclic voltammetry to survey dopamine release in the striatum and analyzed the rats’ behavior using novel object recognition (NOR) androtarod tests. Both tonic and bursting dopamine release were greatly depressed in the severely (6 Psi) injured group, which persisted up to 8 weeks later. However, in the 2 Psi-injured group, the suppression of bursting dopamine release occurred at 1~2 weeks after injury, but there were no significant differences after 4 weeks. Tonic dopamine release was also diminished significantly at 1~2 weeks after the injury; partial recovery could then be seen 4 weeks after injury. A significant deficiency in the fixed speedrotarod test and NOR test were noted in both 2 Psi and 6 Psi groups initially; however, the changes recovered in the 2 Psi group 2 weeks after injury while persisting in the 6 Psi group. In conclusion, striatal evoked dopamine release was affected by fluid percussion injury, with behavioral deficits showing differences as a function of injury severity. The severe fluid percussion injury (6 Psi) group showed more dopamine release defects, as well as cognitive and motor deficiencies. Recovery of dopamine release and improvement in behavioral impairment were better in the mild TBI group.

Key words: Fluid percussion injury; Dopamine; Fast-scan cyclic voltammetry (FSCV); Novel object recognition (NOR) test; Dopamine reuptake

Received April 30, 2014; final acceptance August 1, 2014. Online prepub date: August 5, 2014.
Address correspondence to Yuan-Hao Chen, M.D., Ph.D., Department of Neurological Surgery, Tri-Service General Hospital, National Defense Medical Center, 4F, No. 325, 2nd Sec., Cheng-Kung Road, Nei-Hu District, Taipei City, 114 Taiwan, R.O.C. Tel: (+886) 2- 8792-7177; Fax: (+886) 2-8792-7178; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2129-2142, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683575
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

CD8lowCD28 T Cells: A Human CD8 T-Suppressor Subpopulation With Alloantigen Specificity Induced by Soluble HLA-A2 Dimer In Vitro

Zhigang Wang,*†1 Lichen Ouyang,*1 Zhihui Liang,* Jun Chen,* Qian Yu,* Victor Tunje Jeza,* Yeli Gong,* Guanxin Shen,* Xiufang Weng,* and Xiongwen Wu*

*Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
†Department of Oncology, Wuhan General Hospital of Guangzhou Command, Wuhan, China

CD8+ suppressor T cells have been demonstrated to provide protection of allografts from rejection. We previously reported that soluble peptide/HLA-A2 dimer shows peptide-specific inhibitory effects on alloresponse in a coculture of peptide-pulsed T2 cells with HLA-A2 negative lymphocytes in vitro. Here we found a subset of CD8lowCD28 T cells that was induced in the dimer-treated coculture. Importantly, this population showed hyporesponsiveness to the alloantigen restimulation as well as alloantigen-specific suppression on alloreactive T cells in a cell–cell contact-dependent fashion. The suppressive mechanisms of CD8lowCD28 T cells involved an elevated expression of membrane-bound TGF-β1, but not Foxp3, CTLA-4, or IL-10. Furthermore, anoverrepresention of CD8lowCD28 T cells was observed in the patients after allogeneic platelet transfusion and positively correlated with the elevated concentrations of plasma HLA class I antigens. Our findings demonstrated that soluble HLA-A2 dimer could efficiently induce the tolerant CD8lowCD28 T cells with alloantigen-specific suppression on alloreactive T cells. This study might provide a new strategy for preparation of donor-specific suppressor T cells and represent an attractive alternative for induction of allograft tolerance.

Key words: Soluble HLA-A2 dimer; Suppressor T cells; CD8lowCD28 T cells; Membrane-bound TGF-β1; Allograft tolerance

Received May 10, 2012; final acceptance July 31, 2014. Online prepub date: August 5, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Prof. Xiongwen Wu, Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Rd, Wuhan 430030, China. Tel: +86-27-83692611; Fax: +86-27-83693500; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  orDr. Xiufang Weng, Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Rd, Wuhan 430030, China. Tel: +86-27-83692611; Fax: +86-27-83693500; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2143-2154, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685249
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

CCL22 Prevents Rejection of Mouse Islet Allografts and Induces Donor-Specific Tolerance

Joel Montane,* Merce Obach,* Sigrid Alvarez,* Loraine Bischoff,* Derek L. Dai,* Galina Soukhatcheva,* John J. Priatel,* Gijs Hardenberg,† Megan K. Levings,† Rusung Tan,* Paul C. Orban,* and C. Bruce Verchere*†

*Department of Pathology and Laboratory Medicine, University of British Columbia and Child and Family Research Institute, Vancouver, BC, Canada
†Department of Surgery, University of British Columbia and Child and Family Research Institute, Vancouver, BC, Canada

Manipulation of regulatory T cell (Treg) migration by islet expression of the chemokine CCL22 prevents diabetes in NOD mice and delays recurrent autoimmunity in syngeneic islet transplants. We sought to determine whether attracting Tregs with CCL22 also prevents islet allograft rejection. Isolated Bl/6 mouse islets were transduced overnight with adenovirus expressing CCL22 (Ad-CCL22) downstream of the CMV promoter. Islets were transplanted under the renal capsule of Balb/c recipients made diabetic by streptozotocin. To assess immunologic tolerance, graft-bearing kidneys from recipients of CCL22-expressing islet grafts were removed, and mice received a second transplant of naive islets from the same donor strain or third-party islets into the contralateral kidney. Adenoviral expression of CCL22 conferred prolonged protection of islet allografts in MHC-mismatched, diabetic recipients, maintaining normoglycemia in 75% of recipients for at least 80 days. Increased frequency of Treg cells was observed in islet grafts transduced with Ad-CCL22 compared with untreated grafts. Normoglycemic recipients of CCL22-expressing islet grafts showed complete absence of antidonor antibodies and no lymphocyte proliferation after exposure to donorsplenocytes. After removal of the primary graft at day 80, mice that received a second transplant with untreated islets from the same donor strain did not reject the grafts, suggesting the development of tolerance. Expression of CCL22 recruits Treg cells to transplanted islets, prevents activation of alloreactive T-cells and islet allograft failure and induces alloantigen-specific tolerance. Manipulation of Treg cells by CCL22 in transplanted islets may be a novel therapeutic strategy for diabetes.

Key words: Diabetes mellitus; Islet transplantation; Allograft; Chemokine; Tolerance; Regulatory T cells (Tregs)

Received January 5, 2013; final acceptance October 13, 2014. Online prepub date: October 20, 2014.

Address correspondence to Bruce Verchere, Child and Family Research Institute, 950 West 28th Ave, Vancouver, British Columbia, V5Z 4H4, Canada.

Tel: +1-604-875-2490; Fax: +1-604-875-2373; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2155-2168, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685438
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Pancreatic Islet-Like Three-Dimensional Aggregates Derived From Human Embryonic Stem Cells Ameliorate Hyperglycemia in Streptozotocin-Induced Diabetic Mice

Joong-Hyun Shim,*†1 JongHyun Kim,*1 Jiyou Han,* Su Yeon An,* Yu Jin Jang,* Jeongsang Son,* Dong-Hun Woo,* Suel-Kee Kim,* and Jong-Hoon Kim*

*Laboratory of Stem Cells and Tissue Regeneration, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea
†Department of Oriental Cosmetic Science, Semyung University, Chungbuk, Republic of Korea

We previously reported the in vitro differentiation of human embryonic stem cells (hESCs) into pancreatic endoderm. Here we demonstrate that islet-like three-dimensional (3D) aggregates can be derived from the pancreatic endoderm by optimizing our previous protocol. Sequential treatment with Wnt3a, activin A, and noggin induced a transient upregulation of T and MixL1, followed by increased expression of endodermal genes, including FOXA2SOX17, and CXCR4. Subsequent treatment with retinoic acid highly upregulated PDX1 expression. We also show that inhibition of sonic hedgehog signaling by bFGF/activin βB and cotreatment with VEGF and FGF7 produced many 3D cellular clusters that express both SOX17 and PDX1. We found for the first time that proteoglycans and vimentin+ mesenchymal cells were mainly localized in hESC-derived PDX1+ clusters. Importantly, treatment with chlorate, an inhibitor of proteoglycan sulfation, together with inhibition of Notch signaling significantly increased the expression of Neurog3 and NeuroD1, promoting a transition from PDX1+ progenitor cells toward mature pancreatic endocrine cells. Purified dithizone+ 3D aggregates generated by our refined protocol produced pancreatic hormones and released insulin in response to both glucose and pharmacological drugs in vitro. Furthermore, the islet-like 3D aggregates decreased blood glucose levels and continued to exhibit pancreatic features after transplantation into diabetic mice. Generation of islet-like 3D cell aggregates from human pluripotent stem cells may overcome the shortage of cadaveric donor islets for future cases of clinical islet transplantation.

Key words: Human embryonic stem cells (hESCs); Sodium chlorate; β-Cells; Pancreatic islet; Diabetes mellitus

Received January 2, 2014; final acceptance November 5, 2014. Online prepub date: November 13, 2014.
1These authors provided equal contribution to this work.
Address correspondence to Jong-Hoon Kim, Ph.D., Building 1/Room 304, College of Life Sciences and Biotechnology, Science Campus, Korea University, Anam-dong 5-ga, Sungbuk-goo, Seoul 136-713, Republic of Korea. Tel: +82-2-3290-3007; Fax: +82-2-3290-3507; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it