Cell Transplantation 23(10) Abstracts

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Cell Transplantation, Vol. 23, pp. 1169-11185, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X668618
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Isolation and Characterization of Multipotent Cells From Human Fetal Dermis

Cinzia Maria Chinnici,* Giandomenico Amico,* Marcello Monti,† Stefania Motta,† Rosario Casalone,‡ Sergio Li Petri,§ Marco Spada,§ Bruno Gridelli,*§ and Pier Giulio Conaldi*§

*Fondazione Ri.MED, Regenerative Medicine and Biomedical Technologies Unit, Department of Laboratory Medicine and Advanced Biotechnologies, ISMETT, Palermo, Italy
†Translational Medicine Department, University of Milan Istituto Clinico Humanitas, Milan, Italy
‡SSD Genetica, Azienda Ospedaliera Ospedale di Circolo e Fondazione Macchi Polo, Varese, Italy
§Mediterranean Institute for Transplantation and Advanced Specialized Therapies (ISMETT), Palermo, Italy

We report that cells from human fetal dermis, termed here multipotent fetal dermal cells, can be isolated with high efficiency by using a nonenzymatic, cell outgrowth method. The resulting cell population was consistent with the definition of mesenchymal stromal cells by the International Society for Cellular Therapy. As multipotent fetal dermal cells proliferate extensively, with no loss of multilineage differentiation potential up to passage 25, they may be an ideal source for cell therapy to repair damaged tissues and organs. Multipotent fetal dermal cells were not recognized as targets by T lymphocytes in vitro, thus supporting their feasibility for allogenic transplantation. Moreover, the expansion protocol did not affect the normal phenotype and karyotype of cells. When compared with adult dermal cells, fetal cells displayed several advantages, including a greater cellular yield after isolation, the ability to proliferate longer, and the retention of differentiation potential. Interestingly, multipotent fetal dermal cells expressed the pluripotency marker SSEA4 (90.56 ± 3.15% fetal vs. 10.5 ± 8.5% adult) and coexpressed mesenchymal and epithelial markers (>80% CD90+/CK18+ cells), coexpression lacking in the adult counterparts isolated under the same conditions. Multipotent fetal dermal cells were able to form capillary structures, as well as differentiate into a simple epithelium in vitro, indicating skin regeneration capabilities.

Key words: Human fetal skin; Progenitor cells; Differentiation potential; Skin regeneration

Received December 13, 2012; final acceptance May 18, 2013. Online prepub date: June 13, 2013.
Address correspondence to Cinzia Maria Chinnici, Ph.D., Fondazione Ri.MED, Via E. Tricomi 5, 90127 Palermo, Italy. Tel: +39 0912192496; Fax: +39 0912192422; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1187-11198, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X668654
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Collagenase H Is Crucial for Isolation of Rat Pancreatic Islets

Atsushi Fujio,* Kazutaka Murayama,† Youhei Yamagata,‡ Kimiko Watanabe,§ Takehiro Imura,§ Akiko Inagaki,§ Naomi Ohbayashi,†¶ Hiroki Shima,# Satoshi Sekiguchi,* Keisei Fujimori,* Kazuhiko Igarashi,# Noriaki Ohuchi,* Susumu Satomi,* and Masafumi Goto*§

*Division of Advanced Surgical Science and Technology, Tohoku University, Sendai, Japan
†Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan
‡Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Tokyo, Japan
§New Industry Creation Hatchery Center, Tohoku University, Sendai, Japan
¶Faculty of Pharmacy, Iwaki Meisei University, Iwaki, Japan
#Graduate School of Medicine, Tohoku University, Sendai, Japan

The role(s) of collagenase G (ColG) and collagenase H (ColH) during pancreatic islet isolation remains controversial, possibly due to the enzyme blends used in the previous studies. We herein examined the role of ColG and ColH using highly pure enzyme blends of recombinant collagenase of each subtype. Rat pancreases were digested using thermolysin, together with ColG, ColH, or ColG/ColH (n = 9, respectively). No tryptic-like activity was detected in any components of the enzyme blends. The efficiency of the collagenase subtypes was evaluated by islet yield and function. Immunohistochemical analysis, in vitro collagen digestion assay, and mass spectrometry were also performed to examine the target matrix components of the crucial collagenase subtype. The islet yield was highest in the ColG/ColH group (4,101 ± 460 islet equivalents). A substantial number of functional islets (2,811 ± 581 islet equivalents) was obtained in the ColH group, whereas no islets were retrieved in the ColG group. Mass spectrometry demonstrated that ColH reacts with collagen I and III. In the immunohistochemical analysis, both collagen I and III were located in exocrine tissues, although collagen III expression was more pronounced. The collagen digestion assay showed that collagen III was more effectively digested by ColH than by ColG. The present study reveals that ColH is crucial, while ColG plays only a supporting role, in rat islet isolation. In addition, collagen III appears to be one of the key targets of ColH.

Key words: Islet isolation; Diabetes mellitus; Collagenase; Collagen; Mass spectrometry

Received January 13, 2013; final acceptance June 11, 2013. Online prepub date: June 13, 2013.
Address correspondence to Masafumi Goto, M.D., Ph.D., New Industry Creation Hatchery Center, Tohoku University, 1-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-0872, Japan. Tel: +81 22 717 7895; Fax: +81 22 717 7899; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1199-1211, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667529
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Anakinra and Tocilizumab Enhance Survival and Function of Human Islets During Culture: Implications for Clinical Islet Transplantation

Afaf Sahraoui,*†‡ Kristine Kloster-Jensen,*†‡ Thor Ueland,‡§ Olle Korsgren,¶ Aksel Foss,*†‡ and Hanne Scholz*†‡

*Institute for Surgical Research, Oslo University Hospital Rikshospitalet, Oslo, Norway
†Section for Transplantation Surgery, Oslo University Hospital Rikshospitalet, Oslo, Norway
‡Institute of Clinical Medicine, University in Oslo, Oslo, Norway
§Institute for Internal Medicine, Oslo University Hospital Rikshospitalet, Oslo, Norway
¶Department of Immunology, Genetics and Pathology, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden

Pretreatment culture before islet transplantation represents a window of opportunity to ameliorate the proinflammatory profile expressed by human β-cells in duress. Anakinra (IL-1 receptor antagonist) and tocilizumab (monoclonal IL-6 receptor antibody) are two known anti-inflammatory agents successfully used in the treatment of inflammatory states like rheumatoid arthritis. Both compounds have also been shown to reduce blood glucose and glycosylated hemoglobin in diabetic patients. We therefore sought to evaluate the impact of anakinra and tocilizumab on human β-cells. The islets were precultured with or without anakinra or tocilizumab and then transplanted in a marginal mass model using human islets in immunodeficient mice. Islet viability was evaluated in an in vitro model. The pretreatment culture led to a significantly improved engraftment in treated islets compared to the vehicle. Anakinra and tocilizumab are not toxic to human islets and significantly reduce markers of inflammation and cell death. These results strongly support a pretreatment culture with anakinra and tocilizumab prior to human islet transplantation.

Key words: Anakinra; Tocilizumab; Islet; Transplantation; Culture

Received January 8, 2013; final acceptance April 22, 2013. Online prepub date: April 29, 2013.
Address correspondence to Afaf Sahraoui, Institute for Surgical Research, Oslo University Hospital Rikshospitalet, N-0027 Oslo, Norway. Tel: +47-23073520; Fax: +47-23073630; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1213-1219, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X669734
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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The Use of 1.5-Anhydroglucitol for Monitoring Glycemic Control in Islet Transplant Recipients

Eduardo Moraes Leao Peixoto,* Nujen Colak Bozkurt,*† Shari Messinger,*‡ Maria Isabel del Olmo Garcia,§ Vincenzo Lauriola,*¶ Andrea Corrales,* Eva Herrada,* Camillo Ricordi,*#** and Rodolfo Alejandro*††

*Diabetes Research Institute, Miami, FL, USA
†Department of Endocrinology and Metabolism, Diskapi Yildirim Beyazit Training and Research Hospital, Ankara, Turkey
‡Division of Biostatistics, Department of Epidemiology and Public Health, University of Miami Miller School of Medicine, Miami, FL, USA
§Department of Endocrinology and Nutrition, University Hospital La Fe, Valencia, Spain
¶Facolta di Scienze Motorie, Universita degli Studi di Milano, Milano, Italy
#DeWitt Daughtry Family Department of Surgery, University of Miami Miller School of Medicine, Miami, FL, USA
**Jackson Memorial Hospital-University of Miami Transplant Institute, University of Miami Miller School of Medicine, Miami, FL, USA
††Department of Medicine, University of Miami Miller School of Medicine, Miami, FL, USA

We evaluated whether 1,5-anhydroglucitol (1,5-AG) (GlycoMark®), a test for measuring postprandial glucose and glucose variability, could be a tool for assessing short-term glycemic control in islet cell transplant (ICT) subjects. Data of 21 subjects, with type 1 DM and allogenic islet transplantation, who had concomitant fructosamine, HbA1c, 1,5-AG (n = 85 samples), and capillary glucose self-monitoring measurements (n = 2,979) were analyzed retrospectively at different time points after ICT. A significant negative association was observed between 1,5-AG and HbA1c (p = 0.02), but not with fructosamine. When HbA1c was divided in quartiles as <5.6, 5.6–5.9, 5.9–6.2, and >6.2, a decrease of an estimated 0.70 ± 0.30 μg/ml in 1,5-AG was associated with each quartile of increase in HbA1c (p < 0.0001). There was a significant decline of 1.64 ± 0.3mg/dl in postprandial glucose values for each 1 unit increase in 1,5-AG (p < 0.0001). For those with HbA1c ≥ 6.0% when 1,5-AG was ≥8.15 μg/ml, the mean estimated glucose level was 103.71 ± 3.66 mg/dl, whereas it was 132.12 ± 3.71 mg/dl when 1,5-AG was <8.15 μg/ml. The glucose variability (Glumax − Glumin) in subjects with 1,5-AG <8.15 μg/ml was 46.23 mg/dl greater than the subjects with 1,5-AG ≥8.15 μg/ml (HbA1c ≥ 6.0%). There was no significant association between GlycoMark and glucose variability where HbA1c < 6%. 1,5-AG significantly associated with postprandial glucose levels and glucose variability in ICT recipients with near-normal HbA1c (6.0–6.5%) levels. These findings suggest that 1,5-AG can be used to differentiate those ICT subjects with higher glucose variability despite having near-normal HbA1c. However, prospective studies are needed to evaluate the association between GlycoMark levels and the parameters of graft dysfunction/failure.

Key words: Islet transplantation; GlycoMark; Glucose variability; Glycemic control

Received November 7, 2012; final acceptance June 14, 2013. Online prepub date: June 25, 2013.
Address correspondence to Rodolfo Alejandro, M.D., 1450 NW 10th Avenue, University of Miami Miller School of Medicine, Miami, FL, 33136, USA. Tel: +1-305-243-5321; Fax: +1-305-243-1058; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1221-1227, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X6698663
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Liver Fat Accumulation After Islet Transplantation and Graft Survival

Cristiane Bauermann Leitão,*† Eduardo Moraes Leao Peixoto,* Antonio C. Westphalen,‡ Leonor G. Mireles-Zavala,* Vincenzo Lauriola,* Karina Bernetti,* Andrea Corrales,* Camillo Ricordi,*§¶# and Rodolfo Alejandro*¶

*Diabetes Research Institute, Miami, FL, USA
†Endocrine Division of Hospital de Clínicas de Porto Alegre, RS, Brazil
‡Department of Radiology and Biomedical Imaging, University of California, San Francisco, CA, USA
§DeWitt Daughtry Family Department of Surgery, University of Miami-Miller School of Medicine, Miami, FL, USA
¶Department of Medicine, University of Miami-Miller School of Medicine, Miami, FL, USA
#Jackson Memorial Hospital-University of Miami Transplant Institute, University of Miami-Miller School of Medicine, Miami, FL, USA.

Our objective is to evaluate if there is an association between liver fat accumulation after islet transplantation (ITx) and graft survival. A cohort study was conducted in 34 subjects with type 1 diabetes postallogeneic ITx. Liver fat content was evaluated by magnetic resonance imaging (MRI) (change in liver signal intensity on inphase and opposed-phase images). Kaplan–Meier curves and Cox regression analysis were performed with islet dysfunction duration as the dependent variable and fat liver content as an independent one. Values of p < 0.05 were significant (SSPS®18.0 and MedCalc®12.5). Patients’ mean age was 40 ± 8 years (diabetes duration: 31 ± 12 years; male: 41%). Islet survival did not differ in patients without (51 months, 95% CI 40–62 months) or with steatosis (48 months, 95% CI 38–58 months; p = 0.55) during islet dysfunction period. Nevertheless, survival curves appear to separate late in the follow-up, and after 40 months steatosis was associated with shorter graft survival (p log rank = 0.049). This association remained (RR 23.5, 95% CI 1.1–516.0; p = 0.045) after adjustments for possible confounding factors. In this sample of subjects with type 1 diabetes submitted to ITx, steatosis was not associated with islet failure in the whole cohort. However, in subjects with functional islets after 40 months, a shorter graft survival was observed in those with steatosis during the islet dysfunction period, even after adjustments to variables known to be associated with islet failure.

Key words: Steatosis; Islet transplantation (ITx); Graft survival

Received February 19, 2013; final acceptance June 14, 2013. Online prepub date: June 25, 2013.
Address correspondence to Dr. Rodolfo Alejandro, Diabetes Research Institute, University of Miami-Miller School of Medicine, 1450 NW 10th Avenue (R134), Miami, Florida 33136, USA. Tel.: +1-305-243-6504; Fax: +1-305-243-1058; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1229-1242, 2014
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DOI: http://dx.doi.org/10.3727/096368913X669743
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Neonatal Livers: A Source for the Isolation of Good-Performing Hepatocytes for Cell Transplantation

Laia Tolosa,* Eugenia Pareja-Ibars,† M. Teresa Donato,*‡§ Miriam Cortés,† Silvia López,* Nuria Jiménez,* José Mir,† José V. Castell,*‡§1 and M. José Gómez-Lechón*§1

*Unidad de Hepatología Experimental, Centro de Investigación, Instituto de Investigación Sanitaria La Fe, Valencia, Spain
†Unidad de Transplante Hepático, Hospital La Fe, Valencia, Spain
‡Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, Valencia, Spain
§CIBERehd, FIS, Barcelona, Spain

Hepatocyte transplantation is an alternative therapy to orthotopic liver transplantation for the treatment of liver diseases. However, the supply of hepatocytes is limited given the shortage of organs available to isolate good-functioning quality cells. Neonatal livers may be a potential source alternative to adult livers to obtain good-performing hepatic cells for hepatocyte transplantation, which has not yet been explored profoundly. High-yield preparations of viable hepatocytes were isolated from 1- to 23-day-old liver donors, cryopreserved, and banked. Cell integrity and functional quality assessment were performed after thawing. Neonatal hepatocytes showed better postthawing recovery compared with adult hepatocytes, as shown by the viability values that did not differ significantly from freshly isolated cells, a higher expression of adhesion molecules (β1-integrin, β-catenin, and E-cadherin), better attachment efficiency, cell survival, and a lower number of apoptotic cells. The metabolic performance of thawed hepatocytes has been assessed by ureogenesis and drugmetabolizing capability (cytochrome P450 and UDP-glucuronosyltransferase enzymes). CYP2A6, CYP2C9, CYP2E1, and CYP3A4 activities were found in all cell preparations, while CYP1A2, CYP2B6, CYP2C19, and CYP2D6 activities were detected only in hepatocytes from a few neonatal donors. The expression of UGT1A1 and UGT1A9 (transcripts and protein) was detected in all hepatocyte preparations, while activity was measured only in some preparations, probably due to lack of maturity of the enzymes. However, isoforms UGT1A6 and UGT2B7 showed considerable activity in all preparations. Compared to adult liver, the hepatocyte isolation procedure in neonatal livers also provides thawed cell suspensions with a higher proportion of hepatic progenitor cells (EpCAM+ staining), which could also participate in regeneration of liver parenchyma after transplantation. These results could imply important advantages of neonatal hepatocytes as a source of high-quality cells to improve human hepatocyte transplantation applicability.

Key words: Adhesion molecules; Cryopreservation; Neonatal hepatocytes; Ureogenesis; Cytochrome P450; UDP-glucuronosyltransferase; EpCAM

Received January 24, 2013; final acceptance June 18, 2013. Online prepub date: June 25, 2013.
1These authors provided equal contribution to this work.
Address correspondence to M. J. Gómez-Lechón, Unidad de Hepatología Experimental, Centro de Investigación, Instituto Investigación Sanitaria La Fe (IIS La Fe), Avda de Campanar 21, 46009-Valencia, Spain. Tel: +34 96 1973048; Fax: +34 96 1973018; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1243-1254, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X668645
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Preoperative Hepatocyte Transplantation Improves the Survival of Rats With Nonalcoholic Steatohepatitis-Related Cirrhosis After Partial Hepatectomy

Yukio Nakamura,*† Toru Mizuguchi,* Naoki Tanimizu,† Norihisa Ichinohe,† Hidekazu Ooe,† Masaki Kawamoto,* Makoto Meguro,* Koichi Hirata,* and Toshihiro Mitaka†

*Department of Surgery I, Sapporo Medical University School of Medicine, Sapporo, Japan
†Department of Tissue Development and Regeneration, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan

Liver failure after liver resection for cirrhosis is a critical problem, and no effective therapy except liver transplantation is currently available. The objective of this study was to examine whether hepatocyte transplantation (HT) reduces the poststandard liver resection mortality rate of rats with nonalcoholic steatohepatitis (NASH)-related cirrhosis. Liver resection for hepatocellular carcinoma (HCC) combined with NASH-related cirrhosis has become increasingly common. We developed a rat model of acute liver failure after two-thirds partial hepatectomy (PH) for NASH-related cirrhosis. The mechanism by which HT improved the survival of the model rats was examined in short- and long-term investigations. Female DPPIV recipient F344 rats were fed the choline-deficient l-amino acid (CDAA)-defined diet for 12 weeks. Some of the rats were transplanted with male F344 DPPIV+ rat hepatocytes 24 h before undergoing PH. The overall post-PH survival of each group was evaluated, and short- and long-term pathological and molecular biological evaluations were also performed. Overall survival was significantly longer in the HT group than the non-HT group (7-day survival rates: 46.7% and 7.7%, respectively). Compared with the recipient livers of the non-HT group, numerous Ki-67+ hepatocytes and few TUNEL+ hepatocytes were observed in the livers of the HT group. At 6 months after the HT, the DPPIV+ hepatocytes had partially replaced the recipient liver and formed hepatocyte clusters in the spleen. Preoperative HT might improve the survival of rats with NASH-related cirrhosis after PH by preventing the host hepatocytes from accelerating their growth and falling into apoptosis.

Key words: Hepatocyte transplantation (HT); Liver regeneration; Liver repopulation; Apoptosis; Nonalcoholic steatohepatitis (NASH)

Received March 21, 2012; final acceptance June 6, 2013. Online prepub date: June 13, 2013.
Address correspondence to Toru Mizuguchi, M.D., Ph.D., Department of Surgery I, Sapporo Medical University School of Medicine, S-1, W-16, Chuo-ku, Sapporo, Hokkaido 060-8543, Japan. Tel: +81-11-611-2111, ext. 3281; Fax: +81-11-613-1678; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1255-1266, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670200
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Transplantation of Human Fetal-Derived Neural Stem Cells Improves Cognitive Function Following Cranial Irradiation

Munjal M. Acharya,* Lori-Ann Christie,* Thomas G. Hazel,† Karl K. Johe,† and Charles L. Limoli*

*Department of Radiation Oncology, University of California, Irvine, CA, USA
†Neuralstem, Inc., Rockville, MD, USA

Treatment of central nervous system (CNS) malignancies typically involves radiotherapy to forestall tumor growth and recurrence following surgical resection. Despite the many benefits of cranial radiotherapy, survivors often suffer from a wide range of debilitating and progressive cognitive deficits. Thus, while patients afflicted with primary and secondary malignancies of the CNS now experience longer local regional control and progression-free survival, there remains no clinical recourse for the unintended neurocognitive sequelae associated with their cancer treatments. Multiple mechanisms contribute to disrupted cognition following irradiation, including the depletion of radiosensitive populations of stem and progenitor cells in the hippocampus. We have explored the potential of using intrahippocampal transplantation of human stem cells to ameliorate radiation-induced cognitive dysfunction. Past studies demonstrated the capability of cranially transplanted human embryonic (hESCs) and neural (hNSCs) stem cells to functionally restore cognition in rats 1 and 4 months after cranial irradiation. The present study employed an FDA-approved fetal-derived hNSC line capable of large scale-up under good manufacturing practice (GMP). Animals receiving cranial transplantation of these cells 1 month following irradiation showed improved hippocampal spatial memory and contextual fear conditioning performance compared to irradiated, sham surgery controls. Significant newly born (doublecortin positive) neurons and a smaller fraction of glial subtypes were observed within and nearby the transplantation core. Engrafted cells migrated and differentiated into neuronal and glial subtypes throughout the CA1 and CA3 subfields of the host hippocampus. These studies expand our prior findings to demonstrate that transplantation of fetal-derived hNSCs improves cognitive deficits in irradiated animals, as assessed by two separate cognitive tasks.

Key words: Human fetal-derived neural stem cells; Transplantation; Radiation; Cognition; Hippocampus; Novel place recognition; Fear conditioning

Received February 26, 2013; final acceptance July 9, 2013. Online prepub date: July 17, 2013.
Address correspondence to Prof. Charles L. Limoli, Department of Radiation Oncology, University of California Irvine, Medical Sciences I, Room B-146B, Irvine, CA 92697-2695, USA. Tel: +1-949-824-3053; Fax: +1-949-824-3566; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1267-1278, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X668636
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Single-Cell Suspension Methodology Favors Survival and Vascularization of Fetal Striatal Grafts in the YAC128 Mouse Model of Huntington’s Disease

G. Cisbani,* M. Saint-Pierre,* and F. Cicchetti*†

*Centre de Recherche du CHU de Québec (CHUQ), Québec, QC, Canada
†Département de Psychiatrie et Neurosciences, Université Laval, Québec, QC, Canada

Cell replacement therapies have yielded variable and short-lived benefits in Huntington’s disease (HD) patients. This suboptimal outcome is likely due to the fact that graft survival is compromised long term because grafts are subjected to a host’s microglial inflammatory response, to a lack of adequate trophic support, and possibly to cortical excitotoxicity. However, graft demise may also relate to more straightforward issues such as cell preparation methodology (solid grafts vs. cell suspension). Indeed, we recently reported that solid grafts are poorly revascularized in HD patients transplanted 9 and 12 years previously. To evaluate whether methodological issues relating to cell preparation may have an impact on graft viability, we implanted green fluorescent protein (GFP+) single-cell suspensions of fetal striatal neuronal cells into the striatum of YAC128 HD mice. Postmortem evaluation yielded comparable graft survival in YAC128 mice and their wild-type littermates (noncarrier) at 1 and 3 months posttransplantation. Additionally, the degrees of graft revascularization in the YAC128 and noncarrier mice were similar, with both capillaries and large-caliber vessels observable within the grafted tissue. Furthermore, GFP+ cells interacted well with host blood vessels, indicating integration of the donor cells within the recipient brain. These observations, combined with our recent report of poor revascularization of solid grafts in the HD-transplanted patients, suggest that the success of cell transplantation can be improved by optimizing methodological aspects relating to cell preparation.

Key words: Huntington’s disease (HD); YAC128 mouse model; Single-cell suspension grafts; Vasculature

Received May 6, 2013; final acceptance June 1, 2013. Online prepub date: June 13, 2013.
Address correspondence to Francesca Cicchetti, Ph.D., Centre de Recherche du CHU de Québec (CHUL), Axe Neurosciences, T2-50, 2705, Boulevard Laurier, Québec, QC, Canada, G1V 4G2. Tel: +418-656-4141, ext. 48853; Fax: +418-654-2753; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1279-1291, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667510
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Melatonin Pretreatment Improves the Survival and Function of Transplanted Mesenchymal Stem Cells After Focal Cerebral Ischemia

Yaohui Tang,*†1 Beibei Cai,*†1 Falei Yuan,*† Xiaosong He,*† Xiaojie Lin,*† Jixian Wang,‡ Yongting Wang,*† and Guo-Yuan Yang*†‡

*Neuroscience and Neuroengineering Research Center, Med-X Research Institute, Shanghai, China
†School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China
‡Shanghai Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

Mesenchymal stem cell (MSC) transplantation has been shown to be beneficial in treating cerebral ischemia. However, such benefit is limited by the low survival of transplanted MSCs in an ischemic microenvironment. Previous studies showed that melatonin pretreatment can increase MSC survival in the ischemic kidney. However, whether it will improve MSC survival in cerebral ischemia is unknown. Our study examined the effect of melatonin pretreatment on MSCs under ischemia-related conditions in vitro and after transplantation into ischemic rat brain. Results showed that melatonin pretreatment greatly increased survival of MSCs in vitro and reduced their apoptosis after transplantation into ischemic brain. Melatonin-treated MSCs (MT-MSCs) further reduced brain infarction and improved neurobehavioral outcomes. Angiogenesis, neurogenesis, and the expression of vascular endothelial growth factor (VEGF) were greatly increased in the MT-MSC-treated rats. Melatonin treatment increased the level of p-ERK1/2 in MSCs, which can be blocked by the melatonin receptor antagonist luzindole. ERK phosphorylation inhibitor U0126 completely reversed the protective effects of melatonin, suggesting that melatonin improves MSC survival and function through activating the ERK1/2 signaling pathway. Thus, stem cells pretreated by melatonin may represent a feasible approach for improving the beneficial effects of stem cell therapy for cerebral ischemia.

Key words: Melatonin; Mesenchymal stem cells (MSCs); Ischemia; Survival; Angiogenesis

Received October 24, 2012; final acceptance April 19, 2013. Online prepub date: April 29, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Guo-Yuan Yang, M.D., Ph.D., Neuroscience and Neuroengineering Research Center, Med-X Research Institute and School of Biomedical Engineering, Shanghai Jiao Tong University, 1954 Hua-shan Road, Shanghai 200030, China. Tel: +86-21-62933186; Fax: +86-21-62932302; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Yongting Wang, Ph.D., Neuroscience and Neuroengineering Research Center, Med-X Research Institute and School of Biomedical Engineering, Shanghai Jiao Tong University, 1954 Hua-shan Road, Shanghai 200030, China. Tel: +86-21-62933186; Fax: +86-21-62932302; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1293-1303, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667727
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
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Effects and Safety of Allogenic Mesenchymal Stem Cell Intravenous Infusion in Active Ankylosing Spondylitis Patients Who Failed NSAIDs: A 20-Week Clinical Trial

Peng Wang,*1Yuxi Li,*1 Lin Huang,* Jiewen Yang,† Rui Yang,* Wen Deng,† Biling Liang,‡ Lie Dai,§ Qingqi Meng,* Liangbin Gao,* Xiaodong Chen,‡ Jun Shen,‡ Yong Tang,* Xin Zhang,* Jingyi Hou,* Jichao Ye,* Keng Chen,* Zhaopeng Cai,* Yanfeng Wu,† and Huiyong Shen*

*Department of Orthopedics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, P. R. China
†Center for Biotherapy, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, P. R. China
‡Department of Radiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, P. R. China
§Department of Rheumatology and Immunology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, P. R. China

Our objective was to evaluate the feasibility, safety, and efficacy of intravenous (IV) infusion of allogenic mesenchymal stem cells (MSCs) in ankylosing spondylitis (AS) patients who are refractory to or cannot tolerate the side effects of nonsteroidal anti-inflammatory drugs (NSAIDs). AS patients enrolled in this study received four IV infusions of MSCs on days 0, 7, 14, and 21. The percentage of ASAS20 responders (the primary endpoint) at the fourth week and the mean ASAS20 response duration (the secondary endpoint) were used to assess treatment response to MSC infusion and duration of the therapeutic effects. Ankylosing Spondylitis Disease Activity Score Containing C-reactive Protein (ASDAS-CRP) and other preestablished evaluation indices were also adopted to evaluate the clinical effects. Magnetic resonance imaging (MRI) was performed to detect changes of bone marrow edema in the spine. The safety of this treatment was also evaluated. Thirty-one patients were included, and the percentage of ASAS20 responders reached 77.4% at the fourth week, and the mean ASAS20 response duration was 7.1 weeks. The mean ASDAS-CRP score decreased from 3.6 ± 0.6 to 2.4 ± 0.5 at the fourth week and then increased to 3.2 ± 0.8 at the 20th week. The average total inflammation extent (TIE) detected by MRI decreased from 533,482.5 at baseline to 480,692.3 at the fourth week (p > 0.05) and 400,547.2 at the 20th week (p < 0.05). No adverse effects were noted. IV infusion of MSCs is a feasible, safe, and promising treatment for patients with AS.

Key words: Ankylosing spondylitis (AS); Autoimmune; Mesenchymal stem cells (MSCs); Nonsteroidal anti-inflammatory drugs (NSAIDs)

Received December 13, 2012; final acceptance May 9, 2013. Online prepub date: May 22, 2013.
1These authors provided equal contribution to this work.
Address correspondence to Huiyong Shen, M.D., Ph.D., Department of Orthopedics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 107# Yan Jiang Road West, Guangzhou, Guangdong, P. R. China 510120. Tel: +86 139 2227 6368; Fax: +86 20 8133 2507; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Yanfeng Wu, M.D., Center for Biotherapy, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 107# Yan Jiang Road West, Guangzhou, Guangdong, P. R. China 510120. Tel: +86 136 0904 0860; Fax: +86 020 8133 2612; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1305-1319, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X665602
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Improvement of Cardiac Stem Cell Sheet Therapy for Chronic Ischemic Injury by Adding Endothelial Progenitor Cell Transplantation: Analysis of Layer-Specific Regional Cardiac Function

Sokichi Kamata,* Shigeru Miyagawa,* Satsuki Fukushima,* Satoshi Nakatani,† Atsuhiko Kawamoto,‡ Atsuhiro Saito,* Akima Harada,* Tatsuya Shimizu,§ Takashi Daimon,¶ Teruo Okano,§ Takayuki Asahara,‡ and Yoshiki Sawa*

*Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Japan
†Division of Functional Diagnostics, Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Suita, Japan
‡Division of Vascular Regeneration Therapy, Institute of Biomedical Research and Innovation, Kobe, Japan
§Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, Tokyo, Japan
¶Department of Biostatistics, Hyogo College of Medicine, Nishinomiya, Hyogo, Japan

The transplantation of cardiac stem cell sheets (CSC sheets) is a promising therapeutic strategy for ischemic cardiomyopathy, although potential ischemia in the transplanted area remains a problem. Injected endothelial progenitor cells (EPCs) can reportedly induce angiogenesis in the injected area. We hypothesized that concomitant CSC sheet transplantation and EPC injection might show better therapeutic effects for chronic ischemic injury model than the transplantation of CSC sheets alone. Scaffold-free CSC sheets were generated from human c-kit-positive heart-derived cells. A porcine chronic ischemic injury model was generated by placing an ameroid constrictor around the left coronary artery for 4 weeks. The animals then underwent a sham operation, epicardial transplantation of CSC sheet over the ischemic area, intramyocardial injection of EPCs into the ischemic and peri-ischemic area, or CSC sheet transplantation plus EPC injection. The efficacy of each treatment was then assessed for 2 months. Speckle-tracking echocardiography was used to dissect the layer-specific regional systolic function by measuring the radial strain (RS). The epicardial RS in the ischemic area was similarly greater after treatment with the CSC-derived cell sheets alone (19 ± 5%) or in combination with EPC injection (20 ± 5%) compared with the EPC only (9 ± 4%) or sham (7 ± 1%) treatment. The endocardial RS in the ischemic area was greatest after the combined treatment (14 ± 1%), followed by EPC only (12 ± 1%), compared to the CSC only (11 ± 1%) and sham (9 ± 1%) treatments. Consistently, either epicardial CSC sheet implantation or intramyocardial EPC injection yielded increased capillary number and reduced cardiac fibrosis in the ischemic epicardium or endocardium, respectively. Concomitant EPC injection induced the migration of transplanted CSCs into the host myocardium, leading to further neovascularization and reduced fibrosis in the ischemic endocardium, compared to the CSC sole therapy. Transplantation of CSC sheets induced significant functional recovery of the ischemic epicardium, and concomitant EPC transplantation elicited transmural improvement in chronic ischemic injury.

Key words: Cardiac stem cells (CSCs); Endothelial progenitor cells (EPCs); Chronic ischemic injury; Strain imaging; Left ventricular remodeling

Received July 26, 2012; final acceptance March 17, 2013. Online prepub date: April 3, 2013.
Address correspondence to Professor Yoshiki Sawa, Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan. Tel: +81-6-6879-3154; Fax: +81-6-6879-3163; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it