Cell Transplantation 26(6) Abstracts

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Cell Transplantation, Vol. 26, pp. 925-947, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368917X
694859
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Review

Pig-to-Primate Islet Xenotransplantation: Past, Present, and Future

Zhengzhao Liu,*1 Wenbao Hu,*1 Tian He,* Yifan Dai,† Hidetaka Hara,‡ Rita Bottino,§ David K. C. Cooper,‡ Zhiming Cai,* and Lisha Mou*

*Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People’s Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong, P.R. China
†Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, P.R. China
‡Xenotransplantation Program/Department of Surgery, The University of Alabama at Birmingham, Birmingham, AL, USA
§Institute for Cellular Therapeutics, Allegheny-Singer Research Institute, Pittsburgh, PA, USA

Islet allotransplantation results in increasing success in treating type 1 diabetes, but the shortage of deceased human donor pancreata limits progress. Islet xenotransplantation, using pigs as a source of islets, is a promising approach to overcome this limitation. The greatest obstacle is the primate immune/inflammatory response to the porcine (pig) islets, which may take the form of rapid early graft rejection (the instant blood-mediated inflammatory reaction) or T-cell-mediated rejection. These problems are being resolved by the genetic engineering of the source pigs combined with improved immunosuppressive therapy. The results of pig-to-diabetic nonhuman primate islet xenotransplantation are steadily improving, with insulin independence being achieved for periods >1 year. An alternative approach is to isolate islets within a micro- or macroencapsulation device aimed at protecting them from the human recipient’s immune response. Clinical trials using this approach are currently underway. This review focuses on the major aspects of pig-to-primate islet xenotransplantation and its potential for treatment of type 1 diabetes.

Key words: Type 1 diabetes (T1D); Encapsulation; Instant blood-mediated inflammatory reaction (IBMIR); Islets; Porcine; Xenotransplantation

Received September 16, 2016; final acceptance March 21, 2017. Online prepub date: February 3, 2017.
1These authors provided equal contribution to this work.
Address correspondence to Lisha Mou, Ph.D., Shenzhen Second People’s Hospital, No. 3002 Sungang Road, Futian District, Shenzhen 518035, P.R. China. Tel: (086) 0755-83366388-3230; Fax: (086) 0755-83366388-3230; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 949-965, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368917X
694877
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Review

Using Electrical Stimulation to Enhance the Efficacy of Cell Transplantation Therapies for Neurodegenerative Retinal Diseases: Concepts, Challenges, and Future Perspectives

Abby Leigh Manthey,* Wei Liu,† Zhi Xin Jiang,* Marcus Hiu Kong Lee,* Jian Ji,† Kwok-Fai So,*‡§¶ Jimmy Shiu Ming Lai,*‡ Vincent Wing Hong Lee,* and Kin Chiu*‡§

*Department of Ophthalmology, The University of Hong Kong, Hong Kong SAR, P.R. China
†Tianjin Medical University Eye Hospital, Tianjin, P.R. China
‡Research Centre of Heart, Brain, Hormone and Healthy Aging, The University of Hong Kong, Hong Kong SAR, P.R. China
§State Key Laboratory of Brain and Cognitive Sciences, The University of Hong Kong, Hong Kong SAR, P.R. China
¶GHM Institute of CNS Regeneration, Jinan University, Guangzhou, P.R. China

Disease or trauma-induced loss or dysfunction of neurons in any central nervous system (CNS) tissue will have a significant impact on the health of the affected patient. The retina is a multilayered tissue that originates from the neuroectoderm, much like the brain and spinal cord. While sight is not required for life, neurodegeneration-related loss of vision not only affects the quality of life for the patient but also has societal implications in terms of health care expenditure. Thus, it is essential to develop effective strategies to repair the retina and prevent disease symptoms. To address this need, multiple techniques have been investigated for their efficacy in treating retinal degeneration. Recent advances in cell transplantation (CT) techniques in preclinical, animal, and in vitro culture studies, including further evaluation of endogenous retinal stem cells and the differentiation of exogenous adult stem cells into various retinal cell types, suggest that this may be the most appropriate option to replace lost retinal neurons. Unfortunately, the various limitations of CT, such as immune rejection or aberrant cell behavior, have largely prevented this technique from becoming a widely used clinical treatment option. In parallel with the advances in CT methodology, the use of electrical stimulation (ES) to treat retinal degeneration has also been recently evaluated with promising results. In this review, we propose that ES could be used to enhance CT therapy, whereby electrical impulses can be applied to the retina to control both native and transplanted stem cell behavior/survival in order to circumvent the limitations associated with retinal CT. To highlight the benefits of this dual treatment, we have briefly outlined the recent developments and limitations of CT with regard to its use in the ocular environment, followed by a brief description of retinal ES, as well as described their combined use in other CNS tissues.

Key words: Neurodegeneration; Retina; Electrical stimulation (ES); Combination therapy

Received August 22, 2016; final acceptance March 17, 2017. Online prepub date: February 3, 2017.
Address correspondence to Dr. Kin Chiu, Department of Ophthalmology, The University of Hong Kong, Room 410, Hong Kong Jockey Club Building for Interdisciplinary Research, 5 Sassoon Road, Pokfulam, Hong Kong SAR, P.R. China. Tel: +852-2831-5356; Fax: +852-2817-0491; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Dr. Vincent Lee, Hong Kong Eye Consultants (Central), Melbourne Plaza, Room 2302-2303, 33 Queen’s Road Central, Hong Kong SAR, P.R. China. Tel: +852-2223-6110; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 967-982, 2017
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DOI: https://doi.org/10.3727/096368917X
694994
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Olfactory Ensheathing Cells Inhibit Gliosis in Retinal Degeneration by Downregulation of the Müller Cell Notch Signaling Pathway

Jing Xie,*† Shujia Huo,*† Yijian Li,*† Jiaman Dai,*† Haiwei Xu,*† and Zheng Qin Yin*†

*Southwest Hospital/Southwest Eye Hospital, Third Military Medical University, Chongqing, P.R. China
†Key Lab of Visual Damage, Regeneration and Restoration of Chongqing, Chongqing, P.R. China

Retinal regeneration and self-repair, whether in response to injury or degenerative disease, are severely impeded by glial scar formation by Müller cells (specialized retinal macroglia). We have previously demonstrated that the activation of Müller cells and gliosis in the degenerative retina are significantly suppressed by the subretinal transplantation of a mixture of olfactory ensheathing cells (OECs) and olfactory nerve fibroblasts. However, the underlying molecular mechanism has remained elusive. Here we transplanted purified rat OECs into the subretinal space of pigmented Royal College of Surgeons (RCS) rats, a classic rodent model of retinal degeneration. Using behavioral testing and electroretinography, we confirmed that the grafted OECs preserved the visual function of rats for 8 weeks, relative to vehicle controls (phosphate-buffered saline). Histological evaluation of outer nuclear layer thickness and composition demonstrated that more photoreceptors and ON-bipolar cells were preserved in the retinas of OEC-treated RCS rats than in controls. The grafted OECs migrated into the outer plexiform layer, inner nuclear layer, and inner plexiform layer. They interacted directly with Müller cells in the retina of RCS rats, in three distinct patterns, and secreted matrix metalloproteinases 2 and 3. Previous studies have demonstrated that rat OECs express delta-like ligand (DLL), while Müller cells express Notch3, the receptor for DLL. Here we found that the grafted OECs significantly decreased the expression, by retinal cells, of Notch signaling pathway components (including Notch3, Notch4, DLL1, DLL4, Jagged1, Hes1, and Hes5) 2 weeks after the cell transplantation and that this effect persisted for a further 2 weeks. Based on these findings, we suggest that transplanted OECs inhibit the activation of Müller cells and the associated gliosis, at least partly through suppression of the Notch pathway.

Key words: Retinitis pigmentosaOlfactory ensheathing cells (OECs); Müller cells; Matrix metalloproteinase 2/3; Notch pathway

Received August 3, 2016; final acceptance March 22, 2017. Online prepub date: February 9, 2017.
Address correspondence to Professor Haiwei Xu, Southwest Hospital/Southwest Eye Hospital, Third Military Medical University, Chongqing 400038, P.R. China. Tel: +86-23-68754803; Fax: +86-23-68754401; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Zheng Qin Yin, Southwest Hospital/Southwest Eye Hospital, Third Military Medical University, Chongqing, 400038, P.R. China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 983-1000, 2017
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DOI: https://doi.org/10.3727/096368917X
694697
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Early Subretinal Allograft Rejection Is Characterized by Innate Immune Activity

Kevin P. Kennelly,*† Toby M. Holmes,‡ Deborah M. Wallace,* Cliona O’Farrelly,§¶ and David J. Keegan*†

*School of Medicine, University College Dublin, Dublin, Ireland
†Department of Ophthalmology, Mater Misericordiae University Hospital, Dublin, Ireland
‡School of Clinical Dentistry, University of Sheffield, Sheffield, UK
§School of Biochemistry and Immunology, Trinity College Dublin, Dublin, Ireland
¶School of Medicine, Trinity College Dublin, Dublin, Ireland

Successful subretinal transplantation is limited by considerable early graft loss despite pharmacological suppression of adaptive immunity. We postulated that early innate immune activity is a dominant factor in determining graft survival and chose a nonimmunosuppressed mouse model of retinal pigment epithelial (RPE) cell transplantation to explore this. Expression of almost all measured cytokines by DH01 RPE cells increased significantly following graft preparation, and the neutrophil chemoattractant KC/GRO/CINC was most significantly increased. Subretinal allografts of DH01 cells (C57BL/10 origin) into healthy, nonimmunosuppressed C57BL/6 murine eyes were harvested and fixed at 1, 3, 7, and 28 days postoperatively and subsequently cryosectioned and stained. Graft cells were detected using SV40 large T antigen (SV40T) immunolabeling and apoptosis/necrosis by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Sections were also immunolabeled for macrophage (CD11b and F4/80), neutrophil (Gr1 Ly-6G), and T-lymphocyte (CD3-ε) infiltration. Images captured with an Olympus FV1000 confocal microscope were analyzed using the Imaris software. The proportion of the subretinal bolus comprising graft cells (SV40T+) was significantly p < 0.001) reduced between postoperative day (POD) 3 (90 ± 4%) and POD 7 (20 ± 7%). CD11b+, F4/80+, and Gr1 Ly-6G+
cells increased significantly ( p < 0.05) from POD 1 and predominated over SV40T+ cells by POD 7. Colabeling confocal microscopic analysis demonstrated graft engulfment by neutrophils and macrophages at POD 7, and reconstruction of z-stacked confocal images confirmed SV40T inside Gr1 Ly-6G+ cells. Expression of CD3-ε was low and did not differ significantly between time points. By POD 28, no graft cells were detectable and few inflammatory cells remained. These studies reveal, for the first time, a critical role for innate immune mechanisms early in subretinal graft rejection. The future success of subretinal transplantation will require more emphasis on techniques to limit innate immune-mediated graft loss, rather than focusing exclusively on suppression of the adaptive immune response.

Key words: Retinal transplantation; Innate immunity; Cell transplantation; Retinal pigment epithelium (RPE); Neutrophil; Macrophage

Received February 19, 2015; final acceptance January 20, 2017. Online prepub date: January 20, 2017.
Address correspondence to David Keegan, Department of Ophthalmology, Mater Misericordiae University Hospital, Eccles Street, Dublin 7, Ireland. Tel: +35318307059; Fax: +35318307198; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1001-1016, 2017
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DOI: https://doi.org/10.3727/096368916X
694391
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Human Dental Pulp Stem Cells Are More Effective Than Human Bone Marrow-Derived Mesenchymal Stem Cells in Cerebral Ischemic Injury

Miyeoun Song,*† Jae-Hyung Lee,†‡ Jinhyun Bae,§ Youngmin Bu,§ and Eun-Cheol Kim*

*Department of Oral and Maxillofacial Pathology, Research Center for Tooth and Periodontal Regeneration (MRC), School of Dentistry, Kyung Hee University, Seoul, Republic of Korea
†Department of Life and Nanopharmaceutical Sciences, Kyung Hee University, Seoul, Republic of Korea
‡Department of Maxillofacial Biomedical Engineering, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea
§Department of Herbal Pharmacology, College of Oriental Medicine, Kyung Hee University, Seoul, Republic of Korea

We compared the therapeutic effects and mechanism of transplanted human dental pulp stem cells (hDPSCs) and human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in a rat stroke model and an in vitro model of ischemia. Rats were intravenously injected with hDPSCs or hBM-MSCs 24 h after middle cerebral artery occlusion (MCAo), and both groups showed improved functional recovery and reduced infarct volume versus control rats, but the hDPSC group showed greater reduction in infarct volume than the hBM-MSC group. The positive area for the endothelial cell marker was greater in the lesion boundary areas in the hDPSC group than in the hBM-MSC group. Administration of hDPSCs to rats with stroke significantly decreased reactive gliosis, as evidenced by the attenuation of MCAo-induced GFAP+/nestin+
and GFAP+/Musashi-1+ cells, compared with hBM-MSCs. In vivo findings were confirmed by in vitro data illustrating that hDPSCs showed superior neuroprotective, migratory, and in vitro angiogenic effects in oxygen–glucose deprivation (OGD)-injured human astrocytes (hAs) versus hBM-MSCs. Comprehensive comparative bioinformatics analyses from hDPSC- and hBM-MSC-treated in vitro OGD-injured hAs were examined by RNA sequencing technology. In gene ontology and KEGG pathway analyses, significant pathways in the hDPSC-treated group were the MAPK and TGF-β signaling pathways. Thus, hDPSCs may be a better cell therapy source for ischemic stroke than hBM-MSCs.

Key words: Human dental pulp stem cells (hDPSCs); Stroke; Cell therapy; Oxygen–glucose deprivation (OGD); Intravenous injection

Received December 3, 2015; final acceptance March 29, 2017. Online prepub date: January 20, 2017.
Address correspondence to Eun-Cheol Kim, D.D.S., Ph.D., Professor, Department of Oral and Maxillofacial Pathology, Research Center for Tooth and Periodontal Regeneration (MRC), School of Dentistry, Kyung Hee University, 26 Kyungheedae-roDongdaemun-gu, Seoul 130-701, Republic of Korea. Tel: +82-2-961-0383; Fax: +82-2-960-2324; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1017-1030, 2017
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DOI: https://doi.org/10.3727/096368917X
695010
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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The Timing of Immunomodulation Induced by Mesenchymal Stromal Cells Determines the Outcome of the Graft in Experimental Renal Allotransplantation

Ana Merino,* Elia Ripoll,* Laura de Ramon,* Nuria Bolaños,* Montserrat Goma,† Oriol Bestard,*‡ Nuria Lloberas,* Josep M. Grinyo,*‡ and Juan Torras Ambròs*‡

*Experimental and Translational Laboratory of Nephrology, Bellvitge Biomedical Research Institute (IDIBELL), Hospitalet de Llobregat, Barcelona, Spain
†Pathology Department, Hospital Universitari de Bellvitge, Hospitalet de Llobregat, Barcelona, Spain
‡Nephrology Department, Hospital Universitari de Bellvitge, Hospitalet de Llobregat, Barcelona, Spain

The immunomodulatory characteristics of mesenchymal stromal cells (MSCs) may lead to multifaceted strategies in rejection of organ transplantation. This study was designed to investigate, first, the effect of the donor-type MSCs from Wistarrats on the immune system of immunocompetent Lewis rats and, second, the rejection responses in a renal transplantation model of Wistar to Lewis. In the first experimental model, MSCs from the bone marrow induced a systemic immune response in the immunocompetent Lewis rats, characterized by two different phases. In the initial phase (days 1–3 after MSCs infusion), the main findings were a decrease in the percentage of the main peripheral blood (PB) lymphocyte subpopulations [T cells, B cells, and natural killer (NK) cells], an increase in the FOXP3 MFI in Tregs, and an elevated concentration of circulating proinflammatory cytokines (IL-1β and TNF-α). In the late phase (days 4–6), the percentage of T cells, B cells, and NK cells returned to baseline levels; the concentration of circulating IL-1β and TNF-α decreased; and the level of anti-inflammatory cytokines (IL-10 and IL-4) increased with respect to the initial phase. In the allogeneic kidney transplantation model, rats were randomized into four groups: nontreated, cyclosporine oral administration, and two groups of rats treated with two different schedules of MSC infusion: 4 days (MSCs-4) and 7 days (MSCs-7) before kidney transplantation and in both a further infusion at the day of transplantation. Both MSC treatments decreased the percentage of T, B, and NK cells in PB. Creatinine levels, survival, and histological parameters were better in MSCs-7 than in MSCs-4. We can conclude that MSCs, by themselves, produce changes in the immune system; they do not need a pathological condition to produce immunomodulatory responses. In the renal allograft model, the optimal time schedule for MSC infusion before grafting was 7 days to prevent acute rejection.

Key words: Mesenchymal stromal cells (MSCs); Renal transplantation; In vivo immunomodulation

Received September 2, 2016; final acceptance March 13, 2017. Online prepub date: February 3, 2017.
Address correspondence to Dr. Juan Torras Ambròs, Nephrology Department, Hospital Universitari de Bellvitge and Institut d’Investigació Biomèdica de Bellvitge (IDIBELL), Feixa Llarga s/n. 08907, L’Hospitalet de Llobregat, Barcelona, Spain. Tel: 0034 932607602; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1031-1042, 2017
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DOI: https://doi.org/10.3727/096368917X
694660
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Human Umbilical Cord Mesenchymal Stem Cells Inhibit T Follicular Helper Cell Expansion Through the Activation of iNOS in Lupus-Prone B6.MRL-Faslpr Mice

Zhuoya Zhang,*1 Ruihai Feng,*1 Lingying Niu,* Saisai Huang,* Wei Deng,* Bingyu Shi,* Genhong Yao,* Weiwei Chen,* Xiaojun Tang,* Xiang Gao,† Xuebing Feng,* and Lingyun Sun*†

*Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, P.R. China
†Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing University, Nanjing, P.R. China

The aberrant generation or activation of T follicular helper (Tfh) cells contributes to the pathogenesis of systemic lupus erythematosus (SLE), yet little is known about how these cells are regulated. In this study, we demonstrated that the frequency of Tfh cells was increased in lupus-prone B6.MRL-
Faslpr (B6.lpr) mice and positively correlated to plasma cell proportions and serum total IgG as well as anti-dsDNA antibody levels. Transplantation of mesenchymal stem cells derived from Wharton’s jelly of human umbilical cords (hUCMSCs) ameliorated lupus symptoms in B6.lpr mice, along with decreased percentages of Tfh cells. In vitro studies showed that the differentiation and proliferation of Tfh cells were markedly suppressed by hUC-MSCs. The production of inducible nitric oxide synthase (iNOS) was dramatically upregulated in hUC-MSCs when cocultured with CD4+ T cells directly, while adding the specific inhibitor of iNOS into the coculture system significantly reversed the inhibitory effect of hUC-MSCs on Tfh cell generation. Interestingly, the efficacy of hUC-MSCs in inhibiting Tfh cells was impaired in the Transwell system, with the reduction of iNOS in both mRNA and protein levels. Taken together, our findings suggest that hUC-MSCs could effectively inhibit Tfh cell expansion through the activation of iNOS in lupus-prone B6.lpr mice, which is highly dependent on cell-to-cell contacts.

Key words: T follicular helper cells; Systemic lupus erythematosus (SLE); Mesenchymal stem cells (MSCs); Inducible nitric oxide synthase (iNOS)

Received August 9, 2016; final acceptance March 17, 2017. Online prepub date: January 20, 2017.
1These authors provided equal contribution to this work.
Address correspondence to Professor Lingyun Sun, Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing 210008, P.R. China. Tel: +86-25-6818-2422; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
or Dr. Xuebing Feng, Department of Rheumatology and Immunology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing 210008, P.R. China. Tel: +86-25-6818-2422; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it



CD34 Antigen and the MPL Receptor Expression Defines a Novel Class of Human Cord Blood-Derived Primitive Hematopoietic Stem Cells

Yoshikazu Matsuoka,* Masaya Takahashi,† Keisuke Sumide,* Hiroshi Kawamura,*‡ Ryusuke Nakatsuka,* Tatsuya Fujioka,* and Yoshiaki Sonoda*

*Department of Stem Cell Biology and Regenerative Medicine, Graduate School of Medical Science, Kansai Medical University, Osaka, Japan
†Department of Pediatrics, Kansai Medical University, Osaka, Japan
‡Department of Orthopedic Surgery, Kansai Medical University, Osaka, Japan

In the murine hematopoietic stem cell (HSC) compartment, thrombopoietin (THPO)/MPL (THPO receptor) signaling plays an important role in the maintenance of adult quiescent HSCs. However, the role of THPO/MPL signaling in the human primitive HSC compartment has not yet been elucidated. We have identified very primitive human cord blood (CB)-derived CD34
severe combined immunodeficiency (SCID)-repopulating cells (SRCs) using the intra-bone marrow injection method. In this study, we investigated the roles of the MPL expression in the human primitive HSC compartment. The SRC activities of the highly purified CB-derived 18LinCD34+/−MPL+/− cells were analyzed using NOG mice. In the primary recipient mice, nearly all mice that received CD34+/−MPL+/− cells were repopulated with human CD45+ cells. Nearly all of these mice that received CD34+MPL+/− and CD34MPL cells showed a secondary repopulation. Interestingly, the secondary recipient mice that received CD34+/−MPL cells showed a distinct tertiary repopulation. These results clearly indicate that the CD34+/− SRCs not expressing MPL sustain a long-term (LT) (>1 year) human cell repopulation in NOG mice. Moreover, CD34 SRCs generate CD34+CD38CD90+ SRCs in vitro and in vivo. These findings provide a new concept that CD34MPL SRCs reside at the apex of the human HSC hierarchy.

Key words: CD34; Myeloproliferative leukemia virus (MPL); Human hematopoietic stem cells (HSCs); HSC hierarchy; Cord blood (CB)

Received August 30, 2016; final acceptance March 28, 2017. Online prepub date: November 30, 2016.
Address correspondence to Professor Yoshiaki Sonoda, M.D., Ph.D., Department of Stem Cell Biology and Regenerative Medicine, Graduate School of Medical Science, Kansai Medical University, Hirakata, Osaka 573-1010, Japan. Tel: +81-72-804-2391; Fax: +81-72-804-2399; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1059-1066, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368917X
694778
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Long-Term Follow-Up of Patients After Autologous Bone Marrow Cell Infusion for Decompensated Liver Cirrhosis

Ja Kyung Kim,* Soo-Jeong Kim,* Yuri Kim,* Yong Eun Chung,† Young Nyun Park,‡ Hyun Ok Kim,§ Jin Seok Kim,* Mi-Suk Park,† Isao Sakaida,¶ Do Young Kim,* Jung Il Lee,* Sang Hoon Ahn,* Kwan Sik Lee,* and Kwang-Hyub Han*

*Department of Internal Medicine, Yonsei University College of Medicine, Seoul, South Korea
†Department of Radiology, Yonsei University College of Medicine, Seoul, South Korea
‡Department of Pathology, Yonsei University College of Medicine, Seoul, South Korea
§Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, South Korea
¶Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan

Although several human clinical trials using various bone marrow-derived cell types for cirrhotic or decompensated patients have reported a short-term benefit, long-term follow-up data are limited. We analyzed the long-term clinical outcomes of autologous bone marrow cell infusion (ABMI) for decompensated liver cirrhosis (LC). Patients enrolled in a pilot single-armed ABMI study were followed up more than 5 years. Bone marrow-derived mononuclear cells (BM-MNCs) from decompensated LC were harvested and after processing were infused into a peripheral vein. The laboratory test results and long-term clinical course including liver transplantation (LT), development of cancer, cause of death, and survival after ABMI were analyzed. Nineteen patients were followed up for a median of 66 months after ABMI. Liver function, including serum levels of albumin and Child–Pugh (CP) score, was improved at the 1-year follow-up. Liver volume was significantly greater, cirrhosis was sustained, and collagen content was decreased at the 6-month follow-up. Five years after ABMI, five patients (26.3%) maintained CP class A without LT or death, and five patients (26.3%) had undergone elective LT. Hepatocellular carcinoma (HCC) occurred in five patients (26.3%), and lymphoma and colon cancer occurred in one patient each. Three patients (15.8%) were lost to follow-up at months 22, 31, and 33, respectively, but maintained CP class A until their last follow-up. Five patients expired due to infection. While improved liver function was maintained in some patients for more than 5 years after ABMI, other patients developed HCC. Further studies of long-term follow-up cohorts after cell therapy for LC are warranted.

Key words: Adult stem cells; Bone marrow; Autologous cells; Transplantation; Liver regeneration; Liver cirrhosis (LC)

Received September 19, 2016; final acceptance March 30, 2017. Online prepub date: January 24, 2017.
Address correspondence to Kwang-Hyub Han, M.D., Department of Internal Medicine, Yonsei University College of Medicine, 250 Seongsanno Seodaemun-gu, Seoul 120-752, South Korea. Tel: 82-2-2228-1949; Fax: 82-2-393-6884; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1067-1076, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368916X
694166
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
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Calcium Concentration in Culture Medium as a Nondestructive and Rapid Marker of Osteogenesis

Yohei Tanikake,* Manabu Akahane,† Akira Furukawa,* Yasuaki Tohma,‡ Yusuke Inagaki,* Tsutomu Kira,* and Yasuhito Tanaka*

*Department of Orthopedic Surgery, Nara Medical University, Kashihara, Nara, Japan
†Department of Public Health, Health Management and Policy, Faculty of Medicine, Nara Medical University, Kashihara, Nara, Japan
‡Department of Orthopedic Surgery, National Hospital Organization, Nara Medical Center, Nara, Japan

Artificial bones made of β-tricalcium phosphate (β-TCP) combined with bone marrow-derived mesenchymal stromal cells (BM-MSCs) are used for effective reconstruction of bone defects caused by genetic defects, traumatic injury, or surgical resection of bone tumors. However, the selection of constructs with high osteogenic potential before implantation is challenging. The purpose of this study was to determine whether the calcium concentration in BM-MSC culture medium can be used as a nondestructive and simple osteogenic marker for selecting tissue-engineered grafts constructed using β-TCP and BM-MSCs. We prepared three cell passages of BM-MSCs derived from three 7-week-old, male Fischer 344 rats; the cells were cultured in osteoinductive medium in the presence of β-TCP for 15 days. The medium was replaced with fresh medium on day 1 in culture and subsequently changed every 48 h; it was collected for measurement of osteocalcin secretion and calcium concentration by enzyme-linked immunosorbent assay and X-ray fluorescence spectrometry, respectively. After cultivation, the constructs were implanted subcutaneously into the backs of recipient rats. Four weeks after implantation, the alkaline phosphatase (ALP) activity and osteocalcin content of the constructs were measured. A strong inverse correlation was observed between the calcium concentration in the medium and the ALP activity and osteocalcin content of the constructs, with Pearson’s correlation coefficients of 0.92 and 0.90, respectively. These results indicate that tissue-engineered bone with high osteogenic ability can be selected before implantation based on low calcium content of the culture medium, resulting in successful bone formation after implantation. This nondestructive, simple method shows great promise for assessing the osteogenic ability of tissue-engineered bone.

Key words: Nondestructive; Osteogenic marker; Calcium concentration; Mesenchymal stromal cells; Tissue engineering

Received November 4, 2016; final acceptance March 13, 2017. Online prepub date: November 29, 2016.
Address correspondence to Yohei Tanikake, M.D., Department of Orthopedic Surgery, Nara Medical University, Kashihara, Nara 634-8522, Japan. Tel: +81-0744-22-3051; Fax: +81-0744-22-4121; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1077-1087, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368917X
694886
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Cisplatin-Impaired Adipogenic Differentiation of Adipose Mesenchymal Stem Cells1

Yu-Hsun Chang,*† Hwan-Wun Liu,‡ Tang-Yuan Chu,†§ Yao-Tseng Wen,¶ Rong-Kung Tsai,¶ and Dah-Ching Ding†§#

*Department of Pediatrics, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
†Lab of Stem Cell Research, Department of Research, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
‡Department of Occupational Medicine, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
§Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
¶Institute of Eye Research, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
#Institue of Medical Science, Tzu Chi University, Hualien, Taiwan, R.O.C.

Adipose tissue-derived mesenchymal stem cells (ADSCs) are derived from adipose tissue and can be induced in vitro to differentiate into osteoblasts, chondroblasts, myocytes, neurons, and other cell types. Cisplatin is a commonly used chemotherapy drug for cancer patients. However, the effects of cisplatin on ADSCs remain elusive. This study found that a high concentration of cisplatin affects the viability of ADSCs. First, the IC50
concentration of cisplatin was evaluated. Proliferation of ADSCs, as assessed by the XTT method, decreased immediately after treatment with various concentrations of cisplatin. ADSCs maintained mesenchymal stem cell surface markers after cisplatin treatment, as determined by flow cytometry. Upon differentiation by adding specific reagents, a significant decrease in adipogenic differentiation (by Oil red O staining) and osteogenic differentiation (by Alizarin red staining), and significant chondrogenic differentiation (by Alcian blue staining) were found after cisplatin treatment. Quantitative RT-PCR was also used in evaluating expression of specific genes to confirm differentiation. Finally, ADSCs from one donor who had received cisplatin showed significantly decreased adipogenic differentiation but increased osteogenic differentiation compared with ADSCs derived from one healthy donor. In conclusion, cisplatin affects the viability, proliferation, and differentiation of ADSCs both in vitro and in vivo via certain signaling pathways, such as p53 and Fas/FasL. The differentiation abilities of ADSCs should be evaluated before their transplantation for repairing cisplatin-induced tissue damage.

Key words: Adipose stem cells; Chemotherapy; Cisplatin; Differentiation; Adipogenesis

Received June 13, 2016; final acceptance March 28, 2017. Online prepub date: February 3, 2017.
1This article was originally submitted for the special Pacific Symposium on Stem Cells and Cancer Research (PPSSC) issue of Cell Transplantation, but it was not completed in time to be included in that issue.
Address correspondence to Dah-Ching Ding, M.D., Ph.D., Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, 707 Chung-Yang Road, Section 3, Hualien, Taiwan, R.O.C. Tel: 886-3-8561825; Fax: 886-3-8577161; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1089-1102, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368917X
694831
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Effects of Different Cell-Detaching Methods on the Viability and Cell Surface Antigen Expression of Synovial Mesenchymal Stem Cells

Kunikazu Tsuji,*1 Miyoko Ojima,†1 Koji Otabe,‡ Masafumi Horie,‡ Hideyuki Koga,† Ichiro Sekiya,‡ and Takeshi Muneta

*Department of Cartilage Regeneration, Tokyo Medical and Dental University, Tokyo, Japan
†Department of Joint Surgery and Sports Medicine, Tokyo Medical and Dental University, Tokyo, Japan
‡Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University, Tokyo, Japan

Flow cytometric analysis of cell surface antigens is a powerful tool for the isolation and characterization of stem cells residing in adult tissues. In contrast to the collection of hematopoietic stem cells, the process of enzymatic digestion is usually necessary to prepare mesenchymal stem cells (MSCs) suspensions, which can influence the expression of cell surface markers. In this study, we examined the effects of various cell-detaching reagents and digestion times on the expression of stem cell-related surface antigens and MSC functions. Human MSCs were detached from dishes using four different reagents: trypsin, TrypLE, collagenase, and a nonenzymatic cell dissociation reagent (C5789; Sigma-Aldrich). Following dissociation reagent incubations ranging from 5 to 120 min, cell surface markers were analyzed by flow cytometry. Trypsin and TrypLE quickly dissociated the cells within 5 min, while collagenase and C5789 required 60 min to obtain maximum cell yields. C5789 significantly decreased cell viability at 120 min. Trypsin treatment significantly reduced CD44+, CD55+, CD73+, CD105+, CD140a+, CD140b+, and CD201+
cell numbers within 30 min. Collagenase treatment reduced CD140a expression by 30 min. In contrast, TrypLE treatment did not affect the expression of any cell surface antigens tested by 30 min. Despite the significant loss of surface antigen expression after 60 min of treatment with trypsin, adverse effects of enzymatic digestion on multipotency of MSCs were limited. Overall, our data indicated that TrypLE is advantageous over other cell dissociation reagents tested for the rapid preparation of viable MSC suspensions.

Key words: Mesenchymal stem cells (MSCs); Surface antigen; Cell-detaching method; Viability; Multipotency

Received September 16, 2016; final acceptance March 28, 2017. Online prepub date: January 31, 2017.
1These authors provided equal contribution to this work.
Address correspondence to Kunikazu Tsuji, Ph.D., Associate Professor, Department of Cartilage Regeneration, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. Tel: +81-3-5803-4080; Fax: +81-3-5803-4675; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 26, pp. 1103-1114, 2017
0963-6897/17 $90.00 + .00
DOI: https://doi.org/10.3727/096368917X
694868
E-ISSN 1555-3892
Copyright © 2017 Cognizant, LLC.
Printed in the USA. All rights reserved

Human Muscle Precursor Cells Overexpressing PGC-1α Enhance Early Skeletal Muscle Tissue Formation

Deana Haralampieva,*†‡ Souzan Salemi,* Ivana Dinulovic,§ Tullio Sulser,* Simon M. Ametamey,† Christoph Handschin,§ and Daniel Eberli*‡

*Laboratory for Tissue Engineering and Stem Cell Therapy, Department of Urology, University Hospital Zurich and University of Zürich, Zürich, Switzerland
†Institute of Pharmaceutical Sciences, ETH Zürich, Zürich, Switzerland
‡Zurich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland
§Biozentrum, Focal Area Growth and Development, University of Basel, Basel, Switzerland

Muscle precursor cells (MPCs) are activated satellite cells capable of muscle fiber reconstruction. Therefore, autologous MPC transplantation is envisioned for the treatment of muscle diseases. However, the density of MPCs, as well as their proliferation and differentiation potential, gradually declines with age. The goals of this research were to genetically modify human MPCs (hMPCs) to overexpress the peroxisome proliferator-activated receptor γ coactivator (PGC-1α), a key regulator of exercise-mediated adaptation, and thereby to enhance early skeletal muscle formation and quality. We were able to confirm the sustained myogenic phenotype of the genetically modified hMPCs. While maintaining their viability and proliferation potential, PGC-1α-modified hMPCs showed an enhanced myofiber formation capacity in vitro. Engineered muscle tissues were harvested 1, 2, and 4 weeks after subcutaneous injection of cell–collagen suspensions, and histological analysis confirmed the earlier myotube formation in PGC-1α-modified samples, predominantly of slow-twitch myofibers. Increased contractile protein levels were detected by Western blot. In summary, by genetically modifying hMPCs to overexpress PGC-1α, we were able to promote early muscle fiber formation in vitro and in vivo, with an initial switch to slow-type myofibers. Therefore, overexpressing PGC-1α is a novel strategy to further enhance skeletal muscle tissue engineering.

Key words: Human muscle precursor cells (hMPCs); Differentiation; Skeletal muscle tissue engineering; Peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α)

Received May 17, 2016; final acceptance March 21, 2017. Online prepub date: Februaury 3, 2017.
Address correspondence to Daniel Eberli, M.D., Ph.D., University Hospital Zurich, Frauenklinikstr. 10, CH-8091 Zürich, Switzerland. Tel: +41 44 255 96 30; Fax: +41 44 255 96 20; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it