Cell Transplantation 25(8) Abstracts

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Cell Transplantation, Vol. 25, pp. 1425-1438, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690836
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Review

Cell Transplantation and Neuroengineering Approach for Spinal Cord Injury Treatment: A Summary of Current Laboratory Findings and Review of Literature

Xin-Yi Lin,* Bi-Qin Lai,* Xiang Zeng,* Ming-Tian Che,* Eng-Ang Ling,† Wutian Wu,‡§¶ and Yuan-Shan Zeng*#**††‡‡

*Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-sen University), Ministry of Education, Guangzhou, Guangdong, China
†Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore
‡School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong
§State Key Laboratory of Brain and Cognitive Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong
¶Jinan University–Hong Kong University Joint Laboratory, GHM Institute of CNS Regeneration, Jinan University, Guangzhou, China
#Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
**Institute of Spinal Cord Injury, Sun Yat-sen University, Guangzhou, China
††Co-innovation Center of Neuroregeneration, Nantong, Jiangsu, China
‡‡Guangdong Provincial Key Laboratory of Brain Function and Disease, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China

Spinal cord injury (SCI) can cause severe traumatic injury to the central nervous system (CNS). Current therapeutic effects achieved for SCI in clinical medicine show that there is still a long way to go to reach the desired goal of full or significant functional recovery. In basic medical research, however, cell transplantation, gene therapy, application of cytokines, and biomaterial scaffolds have been widely used and investigated as treatments for SCI. All of these strategies when used separately would help rebuild, to some extent, the neural circuits in the lesion area of the spinal cord. In light of this, it is generally accepted that a combined treatment may be a more effective strategy. This review focuses primarily on our recent series of work on transplantation of Schwann cells and adult stem cells, and transplantation of stem cell-derived neural network scaffolds with functional synapses. Arising from this, an artificial neural network (an exogenous neuronal relay) has been designed and fabricated by us—a biomaterial scaffold implanted with Schwann cells modified by the neurotrophin-3 (NT-3) gene and adult stem cells modified with the TrkC (receptor of NT-3) gene. More importantly, experimental evidence suggests that the novel artificial network can integrate with the host tissue and serve as an exogenous neuronal relay for signal transfer and functional improvement of SCI.

Key words: Neural network;
Exogenous neuronal relay; Neurotrophin-3 (NT-3); Nerve regeneration; Spinal cord injury (SCI)

Received December 7, 2015; final acceptance May 17, 2016. Online prepub date: February 2, 2016.
Address correspondence to Yuan-Shan Zeng, M.D., Ph.D., Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, #74 Zhongshan 2nd Road, Guangzhou 510080, China. Tel: +86-20-87332698; Fax: +86-20-87332698; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1439-1451, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691466
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Neural Stem Cells Restore Hair Growth Through Activation of the Hair Follicle Niche

Insik Hwang,*1 Kyung-Ah Choi,*†1 Hang-Soo Park,* Hyesun Jeong,* Jeong-Ok Kim,‡ Ki-Cheon Seol,‡ Han-Jin Kwon,§ In-Hyun Park,¶ and Sunghoi Hong*

*School of Biosystem and Biomedical Science, College of Health Science, Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea
†Department of Chemistry, College of Science, Korea University, Anam-dong, Seongbuk-gu, Seoul, Republic of Korea
‡Stem Cell Research Center, Future Cell Therapy, Jamwon-dong, Seocho-gu, Seoul, Republic of Korea
§Dermaster Clinic, Cheongdam-dong, Gangnam-gu, Seoul, Republic of Korea
¶Department of Genetics, Stem Cell Center, Yale School of Medicine, Yale University, New Haven, CT, USA

Several types of hair loss result from the inability of hair follicles to initiate the anagen phase of the hair regeneration cycle. Modulating signaling pathways in the hair follicle niche can stimulate entry into the anagen phase. Despite much effort, stem cell-based or pharmacological therapies to activate the hair follicle niche have not been successful. Here, we set out to test the effect of neural stem cell (NSC) extract on the hair follicle niche for hair regrowth. NSC extracts were applied to the immortalized cell lines HaCaT keratinocytes and dermal papilla cells (DPCs) and the shaven dorsal skin of mice. Treatment with NSC extract dramatically improved the growth of HaCaT keratinocytes and DPCs. In addition, NSC extract enhanced the hair growth of the shaven dorsal skin of mice. In order to determine the molecular signaling pathways regulated by NSCs, we evaluated the expression levels of multiple growth and signaling factors, such as insulin-like growth factor-1 (IGF-1), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β), and bone morphogenetic protein (BMP) family members. We found that treatment with an NSC extract enhanced hair growth by activating hair follicle niches via coregulation of TGF-β and BMP signaling pathways in the telogen phase. We also observed activation and differentiation of intrafollicular hair follicle stem cells, matrix cells, and extrafollicular DPCs in vivo and in vitro. We tested whether activation of growth factor pathways is a major effect of NSC treatment on hair growth by applying the growth factors to mouse skin. Combined growth factors, including TGF-β, significantly increased the hair shaft length and growth rate. DNA damage and cell death were not observed in skin cells of mice treated with the NSC extract for a prolonged period. Overall, our data demonstrate that NSC extract provides an effective approach for promoting hair growth by directly regulating hair follicle niches through TGF-β and BMP signaling pathways as well as induction of core growth factors.

Key words: Hair growth; Neural stem cell extract; Hair follicle niche; Transforming growth factor-β (TGF-β)

Received February 9, 2016; final acceptance May 10, 2016. Online prepub date: April 22, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Sunghoi Hong, Ph.D., School of Biosystem and Biomedical Science, College of Health Science, Korea University, 145 Anam-roSeongbuk-gu, Seoul 136-701, Republic of Korea. Tel: +82-2-3290-5636; Fax: +82-2-916-5943; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1453-1460, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690971
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Breaking the Blood–Brain Barrier With Mannitol to Aid Stem Cell Therapeutics in the Chronic Stroke Brain

Naoki Tajiri,1 Jea Young Lee,1 Sandra Acosta,1 Paul R. Sanberg, and Cesar V. Borlongan

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

Blood–brain barrier (BBB) permeabilizers, such as mannitol, can facilitate peripherally delivered stem cells to exert therapeutic benefits on the stroke brain. Although this BBB permeation-aided stem cell therapy has been demonstrated in the acute stage of stroke, such BBB permeation in the chronic stage of the disease remains to be examined. Adult Sprague–Dawley rats initially received sham surgery or experimental stroke via the 1-h middle cerebral artery occlusion (MCAo) model. At 1 month after the MCAo surgery, stroke animals were randomly assigned to receive human umbilical cord stem cells only (2 million viable cells), mannitol only (1.1 mol/L mannitol at 4°C), combined human umbilical cord stem cells (200,000 viable cells) and mannitol (1.1 mol/L mannitol at 4°C), and vehicle (phosphate-buffered saline) only. Stroke animals that received human umbilical cord blood cells alone or combined human umbilical cord stem cells and mannitol exhibited significantly improved motor performance and significantly better brain cell survival in the peri-infarct area compared to stroke animals that received vehicle or mannitol alone, with mannitol treatment reducing the stem cell dose necessary to afford functional outcomes. Enhanced neurogenesis in the subventricular zone accompanied the combined treatment of human umbilical cord stem cells and mannitol. We showed that BBB permeation facilitates the therapeutic effects of a low dose of peripherally transplanted stem cells to effectively cause functional improvement and increase neurogenesis in chronic stroke.

Key words: Cerebral ischemia; Stem cells; Blood–brain barrier (BBB); Neurogenesis

Received November 23, 2015; final acceptance March 2, 2016. Online prepub date: February 15, 2016.
1These authors provided equal contribution to this work.
Address correspondence to Dr. Cesar V. Borlongan, Department of Neurosurgery, University of South Florida, 12901 Bruce B. Downs Blvd, MDC78, Tampa, FL 33612, USA. Tel: 813-974-3154; Fax: 813- 974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1461-1471, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X690279
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Safety and Therapeutic Potential of M2 Macrophages in Stroke Treatment

Elena R. Chernykh,* Ekaterina YaShevela,* Nataliya M. Starostina,† Sergey A. Morozov,† Mariya N. Davydova,† Elena V. Menyaeva,† and Alexandr A. Ostanin*

*Laboratory of Cellular Immunotherapy, Institute of Fundamental and Clinical Immunology, Novosibirsk, Russia
†Clinical Department, Institute of Fundamental and Clinical Immunology, Novosibirsk, Russia

Our objective was to evaluate the safety and clinical efficacy of autologous M2 macrophage transplantation in nonacute stroke patients. We also evaluated whether the intrathecal administration of macrophages influences the production of cytokines by peripheral blood cells and whether the levels of cytokines correlate with stroke severity and responsiveness to cell therapy. In this study, 13 patients (12 males and 1 female with a median age of 63 years) diagnosed with ischemic (n = 10) or hemorrhagic (n = 3) stroke were subjected to cell transplantation therapy (study group). On average, 21.9 × 106
autologous M2 macrophages were injected intrathecally. Thirteen matched case-control stroke patients who did not receive cell therapy comprised the control group. We did not observe any serious adverse events (i.e., intrahospital mortality, neurological worsening, and seizures) related to the cell injection. One patient in the study group and two patients in the control group died during the 6-month follow-up period due to recurrent stroke. In the study group, the NIHSS score decreased from 11 to 6 (p = 0.007) in 6 months after the therapy, whereas the patients in the control group showed a less pronounced neurological improvement (the NIHSS score decreased from 11 to 8, p = 0.07). The obvious positive response (the improvement of the NIHSS score ³3) in the study group was observed in 75% versus 18% in the control group (pFET = 0.03). M2 cell introduction did not significantly affect the production of various cytokines. Nevertheless, pretreated levels of IL-8, IL-10, and IL-4 correlated with stroke severity. Moreover, responder patients had lower spontaneous production of IL-10, FGF-β, PDGF, VEGF, and higher stimulation indexes of IL-1β, TNF-α, IFN-γ, and IL-6 than nonresponders. These findings suggest that the intrathecal administration of autologous M2 cells in stroke patients is safe and leads to a better neurological recovery, which could be mediated through the immunomodulatory activity of M2 macrophages.

Key words: Macrophages; Cell therapy; Nonacute stroke

Received April 14, 2015; final acceptance May 18, 2016. Online prepub date: December 14, 2015.
Address correspondence to Ekaterina Shevela, M.D., Ph.D., Institute of Fundamental and Clinical Immunology, Yadrintsevskaya Street 14, Novosibirsk 630099, Russia. Tel: +7(383)2360329; Fax: +7(383)2227028; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1473-1488, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691277
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Amelioration of Ischemic Brain Injury in Rats With Human Umbilical Cord Blood Stem Cells: Mechanisms of Action

Laura L. Hocum Stone,* Feng Xiao,* Jessica Rotschafer,† Zhenhong Nan,* Mario Juliano,‡ Cyndy D. Sanberg,§ Paul R. Sanberg,¶ Nicole Kuzmin-Nichols,§ Andrew Grande,*‡ Maxim C-J Cheeran,† and Walter C. Low*‡

*Department of Neurosurgery, University of Minnesota, Minneapolis, MN, USA
†Department of Veterinary Population Medicine, University of Minnesota, Minneapolis, MN, USA
‡Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA
§Saneron CCEL Therapeutics Inc., Tampa, FL, USA
¶Center for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, Morsani College of Medicine, University of South Florida, Tampa, FL, USA

Despite the high prevalence and devastating outcome, there remain a few options for treatment of ischemic stroke. Currently available treatments are limited by a short time window for treatment and marginal efficacy when used. We have tested a human umbilical cord blood-derived stem cell line that has been shown to result in a significant reduction in stroke infarct volume as well as improved functional recovery following stroke in the rat. In the present study we address the mechanism of action and compared the therapeutic efficacy of high- versus low-passage nonhematopoietic umbilical cord blood stem cells (nh-UCBSCs). Using the middle cerebral arterial occlusion (MCAo) model of stroke in Sprague–Dawley rats, we administered nh-UCBSC by intravenous (IV) injection 2 days following stroke induction. These human cells were injected into rats without any immune suppression, and no adverse reactions were detected. Both behavioral and histological analyses have shown that the administration of these cells reduces the infarct volume by 50% as well as improves the functional outcome of these rats following stroke for both high- and low-passaged nh-UCBSCs. Flow cytometry analysis of immune cells present in the brains of normal rats, rats with ischemic brain injury, and ischemic animals with nh-UCBSC treatment confirmed infiltration of macrophages and T cells consequent to ischemia and reduction to normal levels with nh-UCBSC treatment. Flow cytometry also revealed a restoration of normal levels of microglia in the brain following treatment. These data suggest that nh-UCBSCs may act by inhibiting immune cell migration into the brain from the periphery and possibly by inhibition of immune cell activation within the brain. nh-UCBSCs exhibit great potential for treatment of stroke, including the fact that they are associated with an increased therapeutic time window, no known ill-effects, and that they can be expanded to high numbers for, and stored for, treatment.

Key words: Stroke; Ischemia; Immune response; Stem cells

Received July 7, 2015; final acceptance March 21, 2016. Online prepub date: March 18, 2016.
Address correspondence to Walter C. Low, Ph.D., Department of Neurosurgery, Stem Cell Institute, University of Minnesota, 2001 6th Street SE, Minneapolis, MN 55455, USA. Tel: +612-626-9203; Fax: +612-626-9201; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1489-1499, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690430
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Stroke Serum Priming Modulates Characteristics of Mesenchymal Stromal Cells by Controlling the Expression miRNA-20a

Eun Hee Kim,*† Dong Hee Kim,†‡ Hye Ree Kim,‡ Soo Yoon Kim,† Hyeon Ho Kim,‡§ and Oh Young Bang†‡¶

*Medical Research Institute, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea
†Translational and Stem Cell Research Laboratory on Stroke, Samsung Medical Center, Seoul, Republic of Korea
‡Department of Health Sciences and Technology, Samsung Advanced Institute for Health Sciences and Technology, Sungkyunkwan University, Seoul, Republic of Korea
§Samsung Biomechanical Research Institute, Institute for Future Medicine, Samsung Medical Center, Seoul, Republic of Korea
¶Department of Neurology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea

Transplantation of mesenchymal stem cells (MSCs) expanded with fetal bovine serum (FBS) has some limitations, including the requirement of a long culture period to obtain a sufficient amount of stem cells. Priming of MSCs with serum from patients with ischemic stroke (stroke serum) increased the proliferation rate and the neurorestorative capacity of MSCs. We hypothesized that this novel priming method increases the proliferation rate of MSCs via the regulation of microRNAs (miRs). Thus, we investigated miR profiling in stroke serum-primed MSCs and tested whether the regulation of certain miRs may affect the proliferation rate of rat MSCs. The proliferation rate of MSCs cultured with stroke serum was higher than that of MSCs cultured with normal serum or FBS. Using miR microarray analysis, we compared the miR expression profiles between MSCs cultured in FBS and in stroke serum. Among miRs associated with cell proliferation, miR-20a was most significantly increased. Similarly, miR-20a was increased in MSCs obtained from the bone marrow of stroke rats compared with MSCs from normal rats. Furthermore, the deregulation of miR-20a by the transfection of MSCs with pre-miR-20a or anti-miR-20a was significantly correlated with the increased proliferation rate of MSCs. The overexpression of miR-20a in MSCs cultured in FBS improved the proliferation rate, while the knockdown of endogenous miR-20a decreased the proliferation rate. In addition, miR-20a promoted proliferation by suppressing the expression of p21 cyclin-dependent kinase inhibitor 1 (CDKN1A). A dual-luciferase reporter assay showed that CDKN1A is a target of miR-20a. Our findings indicate that stroke serum priming upregulated the expression of miR-20a, which promoted MSC proliferation by regulating the cell cycle inhibitor p21 CDKN1A, and suggest the possible roles of priming methods in modulating the characteristics of MSCs by controlling the expression of miR in MSCs.

Key words: Mesenchymal stem cells (MSCs); Proliferation; Stroke serum priming; miRNA-20a; Cyclin-dependent kinase inhibitor 1A (CDKN1A)

Received September 8, 2015; final acceptance March 22, 2016. Online prepub date: January 13, 2016.
Address correspondence to Oh Young Bang, M.D., Ph.D., Department of Neurology, Samsung Medical Center, Sungkyunkwan University, 81 Irwon-ro, Gangnam-gu, Seoul 135-710, South Korea. Tel: 82-2-3410-3599; Fax: 82-2-3410-0052; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1501-1514, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690827
E-ISSN 1555-3892
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A Chemically Defined Medium-Based Strategy to Efficiently Generate Clinically Relevant Cord Blood Mesenchymal Stromal Colonies

Mario Barilani,*† Cristiana Lavazza,* Valentina Boldrin,* Enrico Ragni,* Valentina Parazzi,* Mariacristina Crosti,‡ Elisa Montelatici,* Rosaria Giordano,* and Lorenza Lazzari*

*Cell Factory, Unit of Cell Therapy and Cryobiology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy
†Department of Industrial Engineering, University of PadovaPadova, Italy
Istituto Nazionale di Genetica Molecolare (INGM), Milano, Italy

During the last decade it has been demonstrated that mesenchymal progenitors are present and can be isolated also from cord blood (CB). Recently, we managed to set up a standard protocol allowing the isolation of mesenchymal stromal cells (MSCs) with high proliferative potential and multiple differentiation capabilities, whereas the generation rate of MSC-initiating colonies could still be further improved. Herein, we strikingly succeeded in defining some simple and basic culture conditions based on the use of a chemically defined medium that increased the colony isolation efficiency up to almost 80% of processed CB units. Importantly, this result was achieved irrespective of CB unit white blood cell content and time elapsed from delivery, two limiting parameters involved with processing CB units. Thus, this high efficiency is guaranteed without strict selection of the starting material. In addition, since we are profoundly concerned about how different culture conditions can influence cell behavior, we devoted part of this study to in-depth characterization of the established CB-MSC populations to confirm their stemness features in this novel isolation and culture system. Therefore, an extended study of their immunophenotype, including classical pericyticmarkers, and a detailed molecular analysis addressing telomere length and also stemness-related microRNA contribution were performed. In summary, we propose a straightforward, extremely efficient, and reliable approach to isolate and expand thoroughly characterized CB-MSCs, even when poor-quality CB units are the only available source, or there is no space for an isolation to fail.

Key words: Cord blood (CB); Cord blood-derived mesenchymal stromal cells (CB-MSCs); Chemically defined medium; Mesenchymal stromal cells (MSCs); Stemness potency assay

Received October 2, 2015; final acceptance March 29, 2016. Online prepub date: February 2, 2016.
Address correspondence to Lorenza Lazzari, Ph.D., Cell Factory, Unit of Cell Therapy and Cryobiology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milano, Italy. Tel: +39 02 5503 4053; Fax: +39 02 5503 2796; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1515-1523, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X691141
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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A Multicenter Study: North American Islet Donor Score in Donor Pancreas Selection for Human Islet Isolation for Transplantation

Ling-jia Wang,*1 Tatsuya Kin,†1 Doug O’Gorman,† A. M. James Shapiro,† Bashoo Naziruddin,‡ Morihito Takita,‡ Marlon F. Levy,‡ Andrew M. Posselt,§ Gregory L. Szot,§ Omid Savari,* Barbara Barbaro,¶ James McGarrigle,¶ Chun Chieh Yeh,¶ Jose Oberholzer,¶ Ji Lei,# Tao Chen,# Moh Lian,# James F. Markmann,# Alejandro Alvarez,** Elina Linetsky,** Camillo Ricordi,** A. N. Balamurugan,†† Gopalakrishnan Loganathan,†† Joshua J. Wilhelm,†† Bernhard J. Hering,†† Rita Bottino,‡‡ Massimo Trucco,‡‡ Chengyang Liu,§§ Zaw Min,§§ Yanjing Li,§§ Ali Naji,§§ Luis A. Fernandez,¶¶ MartynasZiemelis,¶¶ Juan S. Danobeitia,¶¶ J. Michael Millis,* and Piotr Witkowski*

*Department of Surgery, Section of Transplantation, University of Chicago, Chicago, IL, USA
†Clinical Islet Transplant Program, University of Alberta and Alberta Health Services, Edmonton, Alberta, Canada
‡Baylor Simmons Transplant Institute, Dallas, TX, USA
§UCSF Transplantation Surgery, University of California-San Francisco, San Francisco, CA, USA
¶UIC Cell Isolation Program, University of Illinois at Chicago, Chicago, IL, USA
#Center for Transplantation Sciences, Massachusetts General Hospital, Boston, MA, USA
**Diabetes Research Institute, cGMP Cell Processing Facility, University of Miami Miller School of Medicine, Miami, FL, USA
††Schulze Diabetes Institute, University of Minnesota, Minneapolis, MN, USA
‡‡Institute of Cellular Therapeutics, Allegheny Health Network, Pittsburgh, PA, USA
§§Division of Transplantation, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA
¶¶Division of Organ Transplantation, University of Wisconsin, Madison, WI, USA

Selection of an optimal donor pancreas is the first key task for successful islet isolation. We conducted a retrospective multicenter study in 11 centers in North America to develop an islet donor scoring system using donor variables. The data set consisting of 1,056 deceased donors was used for development of a scoring system to predict islet isolation success (defined as postpurification islet yield >400,000 islet equivalents). With the aid of univariate logistic regression analyses, we developed the North American Islet Donor Score (NAIDS) ranging from 0 to 100 points. The c index in the development cohort was 0.73 (95% confidence interval 0.70–0.76). The success rate increased proportionally as the NAIDS increased, from 6.8% success in the NAIDS < 50 points to 53.7% success in the NAIDS ≥ 80 points. We further validated the NAIDS using a separate set of data consisting of 179 islet isolations. A comparable outcome of the NAIDS was observed in the validation cohort. The NAIDS may be a useful tool for donor pancreas selection in clinical practice. Apart from its utility in clinical decision making, the NAIDS may also be used in a research setting as a standardized measurement of pancreas quality.

Key words: Islet isolation; Islet transplantation; Organ donor, Pancreas

Received October 5, 2015; final acceptance March 23, 2016. Online prepub date: February 26, 2016.
1These authors are equal first authors.
Address correspondence to Piotr Witkowski, The University of Chicago Medical Center, Department of Surgery, Division of Abdominal Organ Transplantation, 5841 S. Maryland Ave. MC5027, Room J-517, Chicago, IL 60637, USA. Tel: (773) 702-6319; Fax: (773) 702-2126; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1525-1537, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690854
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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Human Fibroblast Sheet Promotes Human Pancreatic Islet Survival and Function In Vitro

Hajime Matsushima,* Tamotsu Kuroki,* Tomohiko Adachi,* Amane Kitasato,* Shinichiro Ono,* Takayuki Tanaka,* Masataka Hirabaru,* Naoki Kuroshima,* Takanori Hirayama,* Yusuke Sakai,* Akihiko Soyama,* Masaaki Hidaka,* Mitsuhisa Takatsuki,* Tatsuya Kin,† James Shapiro,† and Susumu Eguchi*

*Department of Surgery, Nagasaki University, Graduate School of Biomedical Sciences, Sakamoto, Nagasaki, Japan
†Clinical Islet Laboratory and Clinical Islet Transplant Program, University of Alberta, Alberta, Canada

In previous work, we engineered functional cell sheets using bone marrow-derived mesenchymal stem cells (BM-MSCs) to promote islet graft survival. In the present study, we hypothesized that a cell sheet using dermal fibroblasts could be an alternative to MSCs, and then we aimed to evaluate the effects of this cell sheet on the functional viability of human islets. Fibroblast sheets were fabricated using temperature-responsive culture dishes. Human islets were seeded onto fibroblast sheets. The efficacy of the fibroblast sheets was evaluated by dividing islets into three groups: the islets-alone group, the coculture with fibroblasts group, and the islet culture on fibroblast sheet group. The ultrastructure of the islets cultured on each fibroblast sheet was examined by electron microscopy. The fibroblast sheet expression of fibronectin (as a component of the extracellular matrix) was quantified by Western blotting. After 3 days of culture, islet viabilities were 70.2 ± 9.8%, 87.4 ± 5.8%, and 88.6 ± 4.5%, and survival rates were 60.3 ± 6.8%, 65.3 ± 3.0%, and 75.8 ± 5.6%, respectively. Insulin secretions in response to high-glucose stimulation were 5.1 ± 1.6, 9.4 ± 3.8, and 23.5 ± 12.4 μIU/islet, and interleukin-6 (IL-6) secretions were 3.0 ± 0.7, 5.1 ± 1.2, and 7.3 ± 1.0 ng/day, respectively. Islets were found to incorporate into the fibroblast sheets while maintaining a three-dimensional structure and well-preserved extracellular matrix. The fibroblast sheets exhibited a higher expression of fibronectin compared to fibroblasts alone. In conclusion, human dermal fibroblast sheets fabricated by tissue-engineering techniques could provide an optimal substrate for human islets, as a source of cytokines and extracellular matrix.

Key words: Islets; Fibroblasts; Cell sheet; Tissue engineering; Cell transplantation

Received September 4, 2015; final acceptance May 16, 2016. Online prepub date: February 2, 2016.
Address correspondence to Susumu Eguchi, Department of Surgery, Nagasaki University, Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan. Tel: +81-95-8197316; Fax: +81-95-8197319; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1539-1546, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689929
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
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High Seeding Density Induces Local Hypoxia and Triggers a Proinflammatory Response in Isolated Human Islets

Daniel Brandhorst,*† Heide Brandhorst,*† Niamh Mullooly,*† Samuel Acreman,*† and Paul R. V. Johnson*†‡

*Nuffield Department of Surgical Sciences, University of Oxford, Headington, Oxford, UK
†Oxford Centre for Diabetes, Endocrinology and Metabolism Headington, Oxford, UK
‡Oxford NIHR Biomedical Research Centre, Oxford, UK

Hypoxia is the main threat to morphological and functional integrity of isolated pancreatic islets. Lack of oxygen seems to be of particular importance for functionality of encapsulated islets. The present study was initiated as an experimental model for the environment experienced by human islets in a confined space present during culture, shipment, and in an implanted macrodevice. Quadruplicate aliquots of isolated human islets (n = 12) were cultured for 24 h at 37°C under normoxic conditions using 24-well plates equipped with 8-μm pore size filter inserts and filled with islet aliquots adjusted to obtain a seeding density of 75, 150, 300, or 600 IEQ/cm
2. After culture viability, glucose-stimulated insulin release, DNA content as well as Bax and Bcl-2 gene expression were measured. Culture supernatants were collected to determine production of VEGF and MCP-1. Viability correlated inversely with IEQ seeding density (r = −0.71, p < 0.001), while the correlation of VEGF and MCP-1 secretion with seeding density was positive (r = 0.78, p < 0.001; r = 0.54, p < 0.001). Decreased viability corresponded with a significant increase in the Bax/Bcl-2 mRNA ratio at 300 and 600 IEQ/cm2 and with a sigificantly reduced glucose-stimulated insulin secretion and insulin content compared to 75 or 150 IEQ/cm2 (p < 0.01). The present study demonstrates that the seeding density is inversely correlated with islet viability and in vitro function. This is associated with a significant increase in VEGF and MCP-1 release suggesting a hypoxic and proinflammatory islet microenvironment.

Key words: Human islet culture; Islet macroencapsulation; Seeding density; Hypoxia; Inflammation

Received June 16, 2015; final acceptance March 22, 2016. Online prepub date: November 20, 2015.
Address correspondence to Heide Brandhorst, Nuffield Department of Surgical Sciences, Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Drive, Oxford OX3 7LE, UK. Tel: +44-1865-8-57252; Fax: +44-1865-8-57299; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1547-1559, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690999
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

The Protective Effect of Transplanting Liver Cells Into the Mesentery on the Rescue of Acute Liver Failure After Massive Hepatectomy

Sadahiko Kita,* Kentaro Yasuchika,* Takamichi Ishii,† Hokahiro Katayama,* Elena Yukie Yoshitoshi,*‡ Satoshi Ogiso,* Takayuki Kawai,* Katsutaro Yasuda,*‡ Ken Fukumitsu,* Masaki Mizumoto,* and Shinji Uemoto*

*Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan
†Department of Surgery, Nishikobe Medical Center, Kobe, Japan
‡Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan

Postoperative liver failure is one of the most critical complications following extensive hepatectomy. Although transplantation of allogeneic hepatocytes is an attractive therapy for posthepatectomy liver failure, transplanting cells via the portal veins typically causes portal vein embolization. The embolization by transplanted cells would be lethal in patients who have undergone massive hepatectomy. Thus, transplant surgeons need to select extrahepatic sites as transplant sites to prevent portal vein embolization. We aimed to investigate the mechanism of how liver cells transplanted into the mesentery protect recipient rats from acute liver failure after massive hepatectomy. We induced posthepatectomy liver failure by 90% hepatectomy in rats. Liver cells harvested from rat livers were transplanted into the mesenteries of hepatectomized rats. Twenty percent of the harvested cells, which consisted of hepatocytes and nonparenchymal cells, were transplanted into each recipient. The survival rate improved significantly in the liver cell transplantation group compared to the control group 7 days after hepatectomy (69 vs. 7%). Histological findings of the transplantation site, in vivo imaging system study findings, quantitative polymerase chain reaction assays of the transplanted cells, and serum albumin measurements of transplanted Nagase analbuminemic rats showed rapid deterioration of viable transplanted cells. Although viable transplanted cells deteriorated in the transplanted site, histological findings and an adenosine-5′-triphosphate (ATP) assay showed that the transplanted cells had a protective effect on the remaining livers. These results indicated that the paracrine effects of transplanted liver cells had therapeutic effects. The same protective effects were observed in the hepatocyte transplantation group, but not in the liver nonparenchymal cell transplantation group. Therefore, this effect on the remnant liver was mainly due to the hepatocytes among the transplanted liver cells. We demonstrated that transplanted liver cells protect the remnant liver from severe damage after massive hepatectomy.

Key words: Liver cell transplantation; Acute liver failure after massive hepatectomy; Mesenteric cell transplantation; Paracrine factor of transplanted liver cells; Remnant liver; Liver nonparenchymal cells

Received August 11, 2015; final acceptance May 17, 2016. Online prepub date: February 15, 2016.
Address correspondence to Kentaro Yasuchika, M.D., Ph.D., Department of Surgery, Graduate School of Medicine, Kyoto University, 54 Kawara-choShogoin, Sakyo-ku, Kyoto 606-8507, Japan. Tel: +81-75-751-3242; Fax: +81-75-751-4246; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1561-1574, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X689776
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

The Orphan Receptor Tyrosine Kinase ROR2 Facilitates MSCs to Repair Lung Injury in ARDS Animal Model

Shi-xia Cai,*†1 Ai-ran Liu,*1 Song Chen,‡ Hong-li He,* Qi-hong Chen,* Jing-yuan Xu,* Chun Pan,* Yi Yang,* Feng-mei Guo,* Ying-zi Huang,* Ling Liu,* and Hai-bo Qiu*

*Department of Critical Care Medicine, Nanjing Zhong-da Hospital, School of Medicine, Southeast University, Nanjing, China
†Department of Critical Care Medicine, The Affiliated Hospital of Qingdao University, Qingdao, China
‡State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, Nanjing, China

There are some limitations to the therapeutic effects of mesenchymal stem cells (MSCs) on acute respiratory distress syndrome (ARDS) due to their low engraftment and differentiation rates in lungs. We found previously that noncanonical Wnt5a signaling promoted the differentiation of mouse MSCs (mMSCs) into type II alveolar epithelial cells (AT II cells), conferred resistance to oxidative stress, and promoted migration of MSCs in vitro. As receptor tyrosine kinase-like orphan receptor 2 (ROR2) is an essential receptor for Wnt5a, it was reasonable to deduce that ROR2 might be one of the key molecules for the therapeutic effect of MSCs in ARDS. The mMSCs that stably overexpressed ROR2 or the green fluorescent protein (GFP) control were transplanted intratracheally into the ARDS mice [induced by intratracheal injection of lipopolysaccharide (LPS)]. The results showed that ROR2-overexpressing mMSCs led to more significant effects than the GFP controls, including the retention of the mMSCs in the lung, differentiation into AT II cells, improvement of alveolar epithelial permeability, improvement of acute LPS-induced pulmonary inflammation, and, finally, reduction of the pathological impairment of the lung tissue. In conclusion, MSCs that overexpress ROR2 could further improve MSC-mediated protection against epithelial impairment in ARDS.

Key words: Mesenchymal stem cells (MSCs); ROR2; Alveolar epithelium injury; Lipopolysaccharide (LPS)

Received March 20, 2015; final acceptance May 10, 2016. Online prepub date: November 3, 2015.
1These authors provided equal contribution to this work.
Address corresponding to Hai-bo Qiu, Department of Critical Care Medicine, Nanjing Zhong-da Hospital, School of Medicine, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China. Tel: +86-25-83272551; Fax: +86-25-83272123; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 25, pp. 1575-1588, 2016
0963-6897/16 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368916X690809
E-ISSN 1555-3892
Copyright © 2016 Cognizant, LLC.
Printed in the USA. All rights reserved

Experimental Support for the Ecoimmunity Theory: Distinct Phenotypes of Nonlymphocytic Cells in SCID and Wild-Type Mice

David E. Ochayon,*1 Boris M. Baranovski,*1 Peter Malkin,* Ronen Schuster,* Noa Kalay,* Rotem Ben-Hamo,† Ido Sloma,† Justin Levinson,* Jared Brazg,* Sol Efroni,† Eli C. Lewis,*2 and Uri Nevo‡2

*Faculty of Health Sciences, Ben-Gurion University of the Negev, Be’er Sheva, Israel
†Faculty of Life Sciences, Bar-Ilan University, Ramat Gan, Israel
‡Department of Biomedical Engineering, The Iby and Aladar Fleischman Faculty of Engineering, Tel Aviv University, Tel Aviv, Israel

Immune tolerance toward “self” is critical in multiple immune disorders. While there are several mechanisms to describe the involvement of immune cells in the process, the role of peripheral tissue cells in that context is not yet clear. The theory of ecoimmunity postulates that interactions between immune and tissue cells represent a predator–prey relationship. A lifelong interaction, shaped mainly during early ontogeny, leads to selection of nonimmune cell phenotypes. Normally, therefore, nonimmune cells that evolve alongside an intact immune system would be phenotypically capable of evading immune responses, and cells whose phenotype falls short of satisfying this steady state would expire under hostile immune responses. This view was supported until recently by experimental evidence showing an inferior endurance of severe combined immunodeficiency (SCID)-derived pancreatic islets when engrafted into syngeneic immune-intact wild-type (WT) mice, relative to islets from WT. Here we extend the experimental exploration of ecoimmunity by searching for the presence of the phenotypic changes suggested by the theory. Immune-related phenotypes of islets, spleen, and bone marrow immune cells were determined, as well as SCID and WT nonlymphocytic cells. Islet submass grafting was performed to depict syngeneic graft functionality. Islet cultures were examined under both resting and inflamed conditions for expression of CD40 and major histocompatibility complex (MHC) class I/II and release of interleukin-1α (IL-1α), IL-1β, IL-6, tumor necrosis factor-α (TNF-α), IL-10, and insulin. Results depict multiple pathways that appear to be related to the sculpting of nonimmune cells by immune cells; 59 SCID islet genes displayed relative expression changes compared with WT islets. SCID cells expressed lower tolerability to inflammation and higher levels of immune-related molecules, including MHC class I. Accordingly, islets exhibited a marked increase in insulin release upon immunocytedepletion, in effect resuming endocrine function that was otherwise suppressed by resident immunocytes. This work provides further support of the ecoimmunity theory and encourages subsequent studies to identify its role in the emergence and treatment of autoimmune pathologies, transplant rejection, and cancer.

Key words: Major histocompatibility complex (MHC) class I; Lymphocytes; Pancreatic islets; Autoimmunity; Ecoimmunity

Received July 18, 2014; final acceptance May 20, 2016. Online prepub date: February 2, 2016.
1These authors provided equal contribution to this work.
2These authors provided equal contribution to this work.
Address correspondence to Uri Nevo, Department of Biomedical Engineering, Tel Aviv University, Ramat Aviv, Tel Aviv 69978, Israel. Tel: 972-3-6407542; Fax: 972-3-6407542; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Eli C. Lewis, Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, PO Box 653, Be’er Sheva 84105, Israel. Tel.: 972-8-6400830; Fax: 972-8-6400830; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it