Cell Transplantation 24(12) Abstracts

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Cell Transplantation, Vol. 24, pp. 2423-2433, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687480
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Bone Marrow Stromal Cells Differentiate Into Corneal Tissue and Prevent Ocular Graft-Versus-Host Disease in Mice

Luis Ignacio Sánchez-Abarca,*†# Emiliano Hernández-Galilea,‡# Rebeca Lorenzo,‡# Carmen Herrero,†# Almudena Velasco,§# Soraya Carrancio,† Teresa Caballero-Velázquez,*† José Ignacio Rodríguez-Barbosa,¶ Marta Parrilla,§ Consuelo Del Cañizo,†# Jesús San Miguel,** José Aijón,§# and José Antonio Pérez-Simón*†

*Department of Hematology, University Hospital Virgen del Rocío/IBIS/CSIC/University of Seville, Seville, Spain
†Department of Hematology, University Hospital of Salamanca, Salamanca, Spain
‡Department of Surgery, Ophthalmology Service, University Hospital of Salamanca, University of Salamanca, Salamanca, Spain
§Department Cell Biology and Pathology, INCyL, University of Salamanca, Salamanca, Spain
Immunobiology Section, Institute of Biomedicine, University of León, León, Spain
#Institute of Biomedicine Investigation of Salamanca (IBSAL), Salamanca, Spain
**Clinica Universidad de Navarra, Centro de Investigación Médica Aplicada (CIMA), Pamplona, Spain

Clinical trials have assessed the use of human bone marrow stromal cells (hBMSCs) for the treatment of immune-related disorders such as graft-versus-host disease (GVHD). In the current study, we show that GFP+-transduced hBMSCs generated from bone marrow migrate and differentiate into corneal tissue after subconjunctival injection in mice. Interestingly, these hBMSCs display morphological features of epithelial, stromal, and endothelial cells and appear at different layers and with different morphologies depending on their position within the epithelium. Furthermore, these cells display ultrastructural properties, such as bundles of intermediate filaments, interdigitations, and desmosomes with GFP
cells, which confirms their differentiation into corneal tissues. GFP+-transduced hBMSCs were injected at different time points into the right eye of lethally irradiated mice undergoing bone marrow transplantation, which developed ocular GVHD (oGVHD). Remarkably, hBMSCs massively migrate to corneal tissues after subconjunctival injection. Both macroscopic and histopathological examination showed minimal or no evidence of GVHD in the right eye, while the left eye, where no hBMSCs were injected, displayed features of GVHD. Thus, in the current study, we confirm that hBMSCs may induce their therapeutic effect at least in part by differentiation and regeneration of damaged tissues in the host. Our results provide experimental evidence that hBMSCs represent a potential cellular therapy to attenuate oGVHD.

Key words: Human bone marrow stromal cells (hBMSCs); Ocular graft-versus-host disease (oGVHD); Cornea; Subconjunctival injection; Cell migration; Differentiation

Received July 22, 2014; final acceptance February 6, 2015. Online prepub date: February 18, 2015.
Address correspondence to José Antonio Pérez-Simón, M.D., Ph.D., Servicio de HematologíaInstituto de Biomedicina de Sevilla (IBIS)/Hospital Universitario Virgen del Rocío/CSIC/Universidad de SevillaAvenida de Manuel Siurot s/n, 41013 Sevilla, Spain. Tel: +34-955-013260; Fax: +34-955-013265; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Emiliano Hernández-Galilea, M.D., Servicio de Oftalmología, Hospital Universitario de Salamanca, Paseo de San Vicente s/n 37007 Salamanca, Spain. Tel: +34-629838631; Fax: +34-923291113; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2435-2448, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687011
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Transplanted Dentate Progenitor Cells Show Increased Survival in an Enriched Environment But Do Not Exert a Neurotrophic Effect on Spatial Memory Within 2 Weeks of Engraftment

Amanda L. Jamal,* Tara L. Walker,† Amanda J. Waber Nguyen,‡ Robert F. Berman,* Gerd Kempermann,† and Ben Waldau*†

*Department of Neurosurgery, University of California, Davis, Sacramento, CA, USA
†Center for Regenerative Therapies Dresden (CRTD), Technische Universitat Dresden, Dresden, Germany
‡Medical Investigation of Neurodevelopmental Disorders (MIND) Institute, University of California, Davis, Sacramento, CA, USA

Cyclin D2 knockout mice show decreased levels of endogenous dentate neurogenesis. We investigated whether transplanted dentate progenitor cells from wild-type mice respond in vivo to an enriched environment and whether they improve deficient dentate neurogenesis through a neurotrophic effect. Adult cyclin D2 knockout mice were transplanted with passaged adult progenitor cells and kept in an enriched environment or under standard housing conditions in isolation. After 1 week, animals living in an enriched environment underwent water maze testing. Progenitor cells grown on a laminin/poly-D-lysine monolayer expressed Sox2 and nestin and could be differentiated in vitro into neurons and astrocytes. After transplantation into the dentate gyrus, cells preferentially survived along the laminin-rich ependymal lining of the basal cistern or basal membrane of capillaries. A subpopulation of transplanted cells migrated into the interstitial space of the hippocampus and was not associated with laminin. Environmental enrichment led to a significant increase in the survival of transplanted progenitor cells on laminin in the dentate gyrus after 2 weeks. However, animals did not show an enhanced performance in the Morris water maze, and transplantation failed to exert a neurotrophic effect on endogenous neurogenesis after 2 weeks. However, a major limitation of the study is the short-term period of investigation, which may have been insufficient to capture functional effects. In conclusion, initial survival of transplanted neural progenitor cells was dependent on the presence of laminin and was significantly enhanced by environmental enrichment. Further studies are needed to address whether an enriched environment continues to promote graft survival over longer periods of time.

Key words: Laminin; Enriched environment; Cyclin D2 knockout; Neurogenesis; Transplantation; Neural progenitor cell

Received February 22, 2014; final acceptance January 21, 2015. Online prepub date: January 23, 2015.
Address correspondence to Ben Waldau, M.D., University of California, Davis, 4860 Y Street, ACC Suite 3740, Sacramento, CA 95817, USA. Tel: +1-916-734-7193; Fax: +1-916-703-5368; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2449-2461, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X679354
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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BDNF Pretreatment of Human Embryonic-Derived Neural Stem Cells Improves Cell Survival and Functional Recovery After Transplantation in Hypoxic–Ischemic Stroke

Sahar Rosenblum,*†‡ Tenille N. Smith,*†‡ Nancy Wang,* Joshua Y. Chua,* Erick Westbroek,* Kendrick Wang,* and Raphael Guzman*†‡

*Department of Neurosurgery, Stanford University School of Medicine, Stanford, CA, USA
†Department of Neurosurgery, University of Basel, Basel, Switzerland
‡Department of Biomedicine, University of Basel, Basel, Switzerland

Intra-arterial neural stem cell (NSC) therapy has the potential to improve long-term outcomes after stroke. Here we evaluate if pretreatment of NSCs with brain-derived neurotrophic factor (BDNF) prior to transplantation improves cell engraftment and functional recovery following hypoxic–ischemic (HI) stroke. Human embryonic-derived NSCs with or without BDNF pretreatment (1 h, 100 ng/ml) were transplanted 3 days after HI stroke. Functional recovery was assessed using the horizontal ladder test. Cell engraftment was evaluated using bioluminescence imaging (BLI) and histological counts of SC121+
cells. Fluoro-Jade C (FJC) and NeuN stains were used to evaluate neuroprotection. The effect of BDNF on NSCs was analyzed using a migration assay, immunocytochemistry, Luminex proteomic assay, and RT-qPCR.BLI analysis demonstrated significantly higher photon flux in the BDNF-treated NSC group compared to untreated NSC (p = 0.049) and control groups (p = 0.0021) at 1 week after transplantation. Immunohistochemistry confirmed increased transplanted cell survival in the cortex (p = 0.0126) and hippocampus (p = 0.0098) of animals injected with BDNF-treated NSCs compared to untreated NSCs. Behavioral testing revealed that the BDNF-treated NSC group demonstrated increased sensorimotor recovery compared to the untreated NSC and control groups (p < 0.001) over the 1-month period (p < 0.001) following transplantation. A significant improvement in performance was found in the BDNF-treated NSC group compared to the control group at 14, 21, and 28 (p < 0.05) days after transplantation. The cortex and hippocampus of the BDNF-treated NSC group had significantly more SC121+ NSCs (p = 0.0125, p = 0.0098), fewer FJC+neurons (p = 0.0370, p = 0.0285), and a higher percentage of NeuN+ expression (p = 0.0354) in the cortex compared to the untreated NSC group. BDNF treatment of NSCs resulted in significantly greater migration to SDF-1, secretion of M-CSF, VEGF, and expression of CXCR4, VCAM-1, Thrombospondins 1 and 2, and BDNF. BDNF pretreatment of NSCs results in higher initial NSC engraftment and survival, increased neuroprotection, and greater functional recovery when compared to untreated NSCs.

Key words: Neural stem cells (NSCs); Brain-derived neurotrophic factor (BDNF); Hypoxic–ischemic (HI) stroke; Intravascular transplantation; Pretreatment

Received September 6, 2012; final acceptance February 18, 2014. Online prepub date: March 3, 2014.
Address correspondence to Raphael Guzman, M.D., Department of Neurosurgery, University of Basel, Spitalstrasse 21, CH-4031 Basel, Switzerland. Tel: +41 61 265 7124; Fax +41 61 265 7138; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2463-2478, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687200
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Placenta-Derived Multipotent Cells (hPDMCs) Modulate Cardiac Injury: From Bench to Small and Large Animal Myocardial Ischemia Studies

Yuan-Hung Liu,* Kai-Yen Peng,†‡ Yu-Wei Chiu,* Yi-Lwun Ho,§ Yao-Horng Wang,¶ Chia-Tung Shun,# Shih-Yun Huang,* Yi-Shuan Lin,* Antoine A. F. de Vries,** Daniel A. Pijnappels,** Nan-Ting Lee,†† B. Linju Yen,† and Men-Luh Yen††‡‡

*Section of Cardiology, Cardiovascular Center, Far Eastern Memorial Hospital, Pan Chiao, New Taipei City, Taiwan
†Regenerative Medicine Research Group, Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan
‡Department of Life Sciences, National Central University, ChungLi, Taiwan
§Department of Internal Medicine, National Taiwan University Hospital (NTUH), Taipei, Taiwan
¶Nursing Department of Yuanpei University, Hsin-Chu City, Taiwan
#Department and Graduate Institute of Forensic Medicine, College of Medicine (COM), National Taiwan University (NTU), Taipei, Taiwan
**Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands
††School of Medicine, COM, Department of Obstetrics/Gynecology, NTU Hospital, NTU, Taipei, Taiwan
‡‡Research Center for Developmental Biology and Regenerative Medicine, NTU, Taipei, Taiwan

Cardiovascular disease is the leading cause of death globally, and stem cell therapy remains one of the most promising strategies for regeneration or repair of the damaged heart. We report that human placenta-derived multipotent cells (hPDMCs) can modulate cardiac injury in small and large animal models of myocardial ischemia (MI) and elucidate the mechanisms involved. We found that hPDMCs can undergo in vitro cardiomyogenic differentiation when cocultured with mouse neonatal cardiomyocytes. Moreover, hPDMCs exert strong proangiogenic responses in vitro toward human endothelial cells mediated by secretion of hepatocyte growth factor, growth-regulated oncogene-a, and interleukin-8. To test the in vivo relevance of these results, small and large animal models of acute MI were induced in mice and minipigs, respectively, by permanent left anterior descending (LAD) artery ligation, followed by hPDMC or culture medium-only implantation with follow-up for up to 8 weeks. Transplantation of hPDMCsinto mouse heart post-acute MI induction improved left ventricular function, with significantly enhanced vascularity in the cell-treated group. Furthermore, in minipigs post-acute MI induction, hPDMC transplantation significantly improved myocardial contractility compared to the control group (p = 0.016) at 8 weeks postinjury. In addition, tissue analysis confirmed that hPDMC transplantation induced increased vascularity, cardiomyogenic differentiation, and antiapoptotic effects. Our findings offer evidence that hPDMCs can modulate cardiac injury in both small and large animal models, possibly through proangiogenesiscardiomyogenesis, and suppression of cardiomyocyte apoptosis. Our study offers mechanistic insights and preclinical evidence on using hPDMCs as a therapeutic strategy to treat severe cardiovascular diseases.

Key words: Human mesenchymal stem cells; Placenta; Myocardial infarction; Pig model; Mouse model; Cardiac differentiation; ProangiogenesisAntiapoptosis

Received March 15, 2014; final acceptance January 11, 2015. Online prepub date: January 23, 2015.
Address correspondence to Men-Luh Yen, M.D., Ph.D., School of Medicine, College of Medicine, Department of Obstetrics/Gynecology, National Taiwan University Hospital; National Taiwan University, No. 1, Jen-Ai Road Section 1, Taipei, Taiwan 100. Tel: +886-2-23123456, ext. 5122; Fax: +886-2-23911302; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2479-2489, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X687799
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Functional and Electrical Integration of Induced Pluripotent Stem Cell-Derived Cardiomyocytes in a Myocardial Infarction Rat Heart

Takahiro Higuchi,* Shigeru Miyagawa,* James T. Pearson,†‡ Satsuki Fukushima,* Atsuhiro Saito,§ Hirotsugu Tsuchimochi,¶ Takashi Sonobe,§ Yutaka Fujii,¶ Naoto Yagi,# Alberto Astolfo,† MikiyasuShirai,¶ and Yoshiki Sawa*

*Department of Cardiac Surgery, Osaka University Graduate School of Medicine, Osaka, Japan
†The Australian Synchrotron, Clayton, Victoria, Australia
‡Monash Biomedical Imaging Facility, Monash University, Clayton, Victoria, Australia
§Medical Center for Translational Research, Osaka University Hospital, Osaka, Japan
¶Department of Cardiac Physiology, National Cerebral and Cardiovascular Center Research Institute, Suita, Osaka, Japan
#SPring-8/JASRI, Sayo, Hyogo, Japan

In vitro expanded beating cardiac myocytes derived from induced pluripotent stem cells (iPSC-CMs) are a promising source of therapy for cardiac regeneration. Meanwhile, the cell sheet method has been shown to potentially maximize survival, functionality, and integration of the transplanted cells into the heart. It is thus hypothesized that transplanted iPSC-CMs in a cell sheet manner may contribute to functional recovery via direct mechanical effects on the myocardial infarction (MI) heart. F344/NJcl-rnu/rnu rats were left coronary artery ligated (n = 30), followed by transplantation of Dsred-labeled iPSC-CM cell sheets of murine origin over the infarct heart surface. Effects of the treatment were assessed, including in vivo molecular/cellular evaluations using a synchrotron radiation scattering technique. Ejection fraction and activation recovery interval were significantly greater from day 3 onward after iPSC-CM transplantation compared to those after sham operation. A number of transplanted iPSC-CMs were present on the heart surface expressing cardiac myosin or connexin 43 over 2 weeks, assessed by immunoconfocal microscopy, while mitochondria in the transplanted iPSC-CMs gradually showed mature structure as assessed by electron microscopy. Of note, X-ray diffraction identified 1,0 and 1,1 equatorial reflections attributable to myosin and actin–myosin lattice planes typical of organized cardiac muscle fibers within the transplanted cell sheets at 4 weeks, suggesting cyclic systolic myosin mass transfer to actin filaments in the transplanted iPSC-CMs. Transplantation of iPSC-CM cell sheets into the heart yielded functional and electrical recovery with cyclic contraction of transplanted cells in the rat MI heart, indicating that this strategy may be a promising cardiac muscle replacement therapy.

Key words: Induced pluripotent stem cells (iPSCs); Regeneration therapy; Cell sheet; Synchrotron imaging

Received August 15, 2014; final acceptance December 20, 2014. Online prepub date: January 20, 2015.
Address correspondence to Professor Yoshiki Sawa, Department of Cardiovascular Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita 565-0871, Japan. Tel: +81668793154; Fax: +81668793163; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2491-2504, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685762
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of Cardiomyocyte Differentiation Potential Between Type 1 Diabetic Donor- and Nondiabetic Donor-Derived Induced Pluripotent Stem Cells

Chika Kikuchi,* Martin Bienengraeber,*† Scott Canfield,*‡ Andrew Koopmeiner,* Richard Schäfer,§ Zeljko J. Bosnjak,*‡ and Xiaowen Bai*‡

*Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, WI, USA
†Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI, USA
‡Department of Physiology, Medical College of Wisconsin, Milwaukee, WI, USA
§Harvard Stem Cell Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA

Type 1 diabetes mellitus (T1DM) is the most common type of diabetes in children and adolescents. Diabetic subjects are more likely to experience a myocardial infarction compared to nondiabetic subjects. In recent years, induced pluripotent stem cells (iPSCs) have received increasing attention from basic scientists and clinicians and hold promise for myocardial regeneration due to their unlimited proliferation potential and differentiation capacity. However, cardiomyogenesis of type 1 diabetic donor-derivediPSCs (T1DM-iPSCs) has not been investigated yet. The aim of the study was to comparatively analyze cardiomyocyte (CM) differentiation capacity of nondiabetic donor-derived iPSCs (N-iPSCs) and T1DM-iPSCs. The differentiated CMs were confirmed by both expression of cardiac-specific markers and presence of cardiac action potential. Since mitochondrial bioenergetics is vital to every aspect of CM function, extracellular acidification rates and oxygen consumption rates were measured using Seahorse extracellular flux analyzer. The results showed that N-iPSCs and T1DMiPSCs demonstrated similar capacity of differentiation into spontaneously contracting CMs exhibiting nodal-, atrial-, or ventricular-like action potentials. Differentiation efficiency was up to 90%. In addition, the CMs differentiated from N-iPSCs and T1DM-iPSCs (N-iPSC-CMs and T1DM-iPSC-CMs, respectively) showed 1) well-regulated glucose utilization at the level of glycolysis and mitochondrial oxidative phosphorylation and 2) the ability to switch metabolic pathways independent of extracellular glucose concentration. Collectively, we demonstrate for the first time that T1DM-iPSCs can differentiate into functional CMs with well-regulated glucose utilization as shown in N-iPSCs, suggesting that T1DM-iPSC-CMs might be a promising autologous cell source for myocardial regeneration in type 1 diabetes patients.

Key words: Cardiomyocytes (CMs); Induced pluripotent stem cells (iPSCs); Bioenergetics; Type 1 diabetes

Received August 19, 2014; final acceptance December 7, 2014. Online prepub date: January 5, 2015.
Address correspondence to Xiaowen Bai, M.D., Ph.D., Department of Anesthesiology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA. Tel: +1-414-955-5755; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2505-2512, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X685320
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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In Vitro Assessment of Human Islet Vulnerability to Instant Blood-Mediated Inflammatory Reaction (IBMIR) and Its Use to Demonstrate a Beneficial Effect of Tissue Culture

Raina D. Ramnath,* Elisa Maillard,* Katherine Jones,* Paul A. Bateman,* Stephen S. J. Hughes,* Jane Gralla,† Paul R. Johnson,* and Derek W. R. Gray*

*University of Oxford, Nuffield Department of Surgical Sciences, John Radcliffe Hospital, Headington, Oxford, England, UK
†Department of Pediatrics, University of Colorado Denver, Aurora, CO, USA

Culture of human pancreatic islets is now routinely carried out prior to clinical islet allotransplantation, using conditions that have been developed empirically. One of the major causes of early islet destruction after transplantation is the process termed instant blood-mediated inflammatory reaction (IBMIR). The aim of this study was to develop in vitro methods to investigate IBMIR and apply them to the culture conditions used routinely in our human islet isolation laboratory. Freshly isolated or precultured (24 h, 48 h) human islets were incubated in either ABO-compatible allogeneic human blood or Hank’s buffered salt solution (HBSS) for 1 h at 37°C. Tissue factor (TF) expression and leukocyte migration were assessed by light microscopy. TF was also quantified by ELISA. To assess b-cell function, glucose-stimulated insulin secretion (GSIS) assay was carried out. The extent of islet b-cell damage was quantified using a proinsulin assay. Islets cultured for 24 h had higher GSIS when compared to freshly isolated or 48-hprecultured islets. Freshly isolated islets had significantly higher TF content than 24-h and 48-h precultured islets. Incubation of freshly isolated human islets in allogeneic human blood released 6.5-fold higher level of proinsulin in comparison to freshly isolated human islets in HBSS. The high level of proinsulin released was significantly attenuated when precultured islets (24 h or 48 h) were exposed to fresh blood. Histological examination of fresh islets in blood clot showed that some islets were fragmented, showing signs of extraislet insulin leakage and extensive neutrophil infiltration and necrosis. These features were markedly reduced when the islets were cultured for 24 h. These results suggest that our standard 24-h islet culture is markedly beneficial in attenuating IBMIR, as evidenced by increased GSIS, lower content of TF, decrease islet fragmentation, and proinsulin release.

Key words: Instant blood-mediated inflammatory reaction (IBMIR); Human islets; Tissue factor (TF); Proinsulin; Glucose-stimulated insulin secretion

Received April 30, 2012; final acceptance October 31, 2014. Online prepub date: November 5, 2014.
Address correspondence to Raina D. Ramnath, Ph.D., University of Bristol, Dorothy Hodgkin Building, Bristol BS1 3NY, UK. Tel: +44-07549495091; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Prof. Derek W. R. Gray, D. Phil (Oxon), F.R.C.S., M.R.C.P., Nuffield Department of Surgical Sciences, Level 6, John Radcliffe Hospital, Headington, Oxford, OX3 9DU, UK. Tel: +44-01865220145; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2513-2525, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686779
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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3D-Printed Drug/Cell Carrier Enabling Effective Release of Cyclosporin A for Xenogeneic Cell-Based Therapy

Tae-Ha Song,*†1 Jinah Jang,*1 Yeong-Jin Choi,‡ Jin-Hyung Shim,§ and Dong-Woo Cho*

*Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Hyoja-dong, Nam-gu, Pohang, Kyungbuk, Korea
†Medical Device Development Center, Daegu-Gyeongbuk Medical Innovation Foundation (DGMIF), Dong-gu, Daegu, Korea
‡Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Hyoja-dong, Nam-gu, Pohang, Kyungbuk, Korea
§Department of Mechanical Engineering, Korea Polytechnic University, SiheungsiGyeonggi-do, Korea

Systemic administration of the immunosuppressive drug cyclosporin A (CsA) is frequently associated with a number of side effects; therefore, sometimes it cannot be applied in sufficient dosage after allogeneic or xenogeneic cell transplantation. Local delivery is a possible solution to this problem. We used 3D printing to develop a CsA-loaded 3D drug carrier for the purpose of local and sustained delivery of CsA. The carrier is a hybrid of CsA-poly(lactic-co-glycolic acid) (PLGA) microsphere-loaded hydrogel and a polymeric framework so that external force can be endured under physiological conditions. The expression of cytokines, which are secreted by spleen cells activated by Con A, and which are related to immune rejection, was significantly decreased in vitro by the released CsA from the drug carrier. Drug carriers seeded with xenogeneic cells (human lung fibroblast) were subcutaneously implanted into the BALB/c mouse. As a result, T-cell-mediated rejection was also significantly suppressed for 4 weeks. These results show that the developed 3D drug carrier can be used as an effective xenogeneic cell delivery system with controllable immunosuppressive drugs for cell-based therapy.

Key words: 3D printing; Drug delivery system; Cyclosporin A; Cell-based therapy; T-cell-mediated rejection

Received July 8, 2014; final acceptance January 12, 2015. Online prepub date: January 20, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Dong-Woo Cho, Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), San 31, Hyoja-dong, Nam-gu, Pohang, Kyungbuk 790-784, Korea. Tel: +82 54 279 2171; Fax: +82 54 279 5419; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2527-2540, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X683566
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Generation of Human Alloantigen-Specific Regulatory T Cells Under Good Manufacturing Practice-Compliant Conditions for Cell Therapy

Mustapha Cheraï,*†1 Yamina Hamel,‡1 Claude Baillou,‡§ Soumia Touil,¶2 Maude Guillot-Delost,‡§2 Frédéric Charlotte,# Laila Kossir,*† Ghislaine Simonin,*† Sébastien Maury,**††‡‡ José L. Cohen,††‡‡§§ and François M. Lemoine*†‡§

*AP-HP, University Hospital La Pitié-Salpêtrière, Department of Biotherapies, Paris, France
†Center of Clinical Investigation in Biotherapies 1420, University Hospital La Pitié-Salpêtrière, Paris, France
‡Sorbonne Universités, UPMC Univ Paris 06, UMR-S CR7, CIMI-Paris, Paris, France
§INSERM, UMR S 1135, CIMI-Paris, Paris, France
¶CNRS, UMR 7211, Immunology Immunopathology and Immunotherapy, Paris, France
#AP-HP, University Hospital La Pitié-Salpêtrière, Department of Pathology, Paris, France
**AP-HP, Henri-Mondor Hospital, Department of Clinical Hematology, Créteil, France
††Université Paris-Est, UMR-S955, UPEC, Créteil, France
‡‡INSERM, U955, Team 21, Créteil, France
§§AP-HP, Henri-Mondor - A. Chenevier Hospital, CIC-BT-504, Créteil, France

Natural regulatory T cells (Tregs) may have a great therapeutic potential to induce tolerance in allogeneic cells and organ transplantations. In mice, we showed that alloantigen-specific Tregs (spe-Tregs) were more efficient than polyclonal Tregs (poly-Tregs) in controlling graft-versus-host disease (GVHD). Here we describe a clinical-grade compliant method for generating human spe-TregsTregs were enriched from leukapheresis products with anti-CD25 immunomagnetic beads, primed twice by allogeneic mature monocyte-derived dendritic cells (mDCs), and cultured during 3 weeks in medium containing interleukin 2 (IL-2), IL-15, and rapamycin. After 3 weeks of culture, final cell products were expanded 8.3-fold from the initial CD25
+ purifications.Immunophenotypic analyses of final cells indicate that they were composed of 88 ± 2.6% of CD4+ T cells, all expressing Treg-specific markers (FOXP3, Helios, GARP, LAP, and CD152). Spe-Tregs were highly suppressive in vitro and also in vivo using axeno-GVHD model established in immunodeficient mice. The specificity of their suppressive activity was demonstrated on their ability to significantly suppress the proliferation of autologous effector T cells stimulated by the same mDCs compared to third-party mDCs. Our data provide evidence that functional alloantigen Tregs can be generated under clinical-grade compliant conditions. Taking into account that 130 × 106 CD25+ cells can be obtained at large scale from standard leukapheresis, our cell process may give rise to a theoretical final number of 1 × 109 spe-Tregs. Thus, using our strategy, we can propose to prepare spe-Tregs for clinical trials designed to control HLA-mismatched GVHD or organ transplantation rejection.

Key words: Regulatory T cells (Tregs); Cell therapy; Alloantigen; Graft-versus-host disease (GVHD); Dendritic cells

Received March 18, 2014; final acceptance July 30, 2014. Online prepub date: August 5, 2014.
1These authors provided equal contribution to this work as first authors.
2These authors provided equal contribution to this work as third authors.
Address correspondence to Professor François M. LemoineGroupe hospitalier Pitié-Salpêtrière, UPMC UMRS CR7, INSERM U1135, 83, Bld de l’Hôpital, 75651 Paris Cedex 13, France. Tel: (+33) 1 42 17 74 42; Fax: (+33) 1 42 17 74 41; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2541-2555, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368914X687020
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Cold Preservation of Human Adult Hepatocytes for Liver Cell Therapy

Cedric Duret,*† Daniel Moreno,‡ Anangi Balasiddaiah,‡1 Solene Roux,*† Phillipe Briolotti,*† Edith Raulet,*† Astrid Herrero,*†§ Helene Ramet,¶ Christine Biron-Andreani,¶ Sabine Gerbal-Chaloin,*† Jeanne Ramos,*†# Francis Navarro,*†§ Jean Hardwigsen,** Patrick Maurel,*† Rafael Aldabe,‡ and Martine Daujat-Chavanieu*†,††

*INSERM, U1040, Institut de Recherche en Biothérapie, F-34295 Montpellier, France
Université Montpellier 1, UMR 1040, F-34295 Montpellier, France
‡Gene Therapy and Hepatology Area, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain
§Department of General and Liver Transplant Surgery, CHU Montpellier Hôpital Saint Eloi, Montpellier, France
¶Hemophilia Treatment Center, CHU Montpellier Hôpital Saint-Eloi, Montpellier, France
#Department of Pathology, CHU Montpellier Hôpital Gui de Chauliac, Montpellier, France
**Department of Digestive Surgery and Liver Transplantation, AP-HM, Hôpital La Conception, Marseille, France
††CHU Montpellier Hôpital Saint-Eloi, Institut de Recherche en Biothérapie, Montpellier, France

Hepatocyte transplantation is a promising alternative therapy for the treatment of hepatic failure, hepatocellular deficiency, and genetic metabolic disorders. Hypothermic preservation of isolated human hepatocytes is potentially a simple and convenient strategy to provide on-demand hepatocytes in sufficient quantity and of the quality required for biotherapy. In this study, first we assessed how cold storage in three clinically safe preservative solutions (UW, HTS-FRS, and IGL-1) affects the viability and in vitro functionality of human hepatocytes. Then we evaluated whether such cold-preserved human hepatocytes could engraft and repopulate damaged livers in a mouse model of liver failure. Human hepatocytes showed comparable viabilities after cold preservation in the three solutions. The ability of fresh and cold-stored hepatocytes to attach to a collagen substratum and to synthesize and secrete albumin, coagulation factor VII, and urea in the medium after 3 days in culture was also equally preserved. Cold-stored hepatocytes were then transplanted in the spleen of immunodeficient mice previously infected with adenoviruses containing a thymidine kinase construct and treated with a single dose of ganciclovir to induce liver injury. Engraftment and liver repopulation were monitored over time by measuring the blood level of human albumin and by assessing the expression of specific human hepatic mRNAs and proteins in the recipient livers by RT-PCR and immunohistochemistry, respectively. Our findings show that cold-stored human hepatocytes in IGL-1 and HTS-FRS preservative solutions can survive, engraft, and proliferate in a damaged mouse liver. These results demonstrate the usefulness of human hepatocyte hypothermic preservation for cell transplantation.

Key words: Cell therapy; Transplantation; Human hepatocytes; Hypothermic preservation; Liver; Engraftment

Received April 7, 2014; final acceptance January 19, 2015. Online prepub date: January 23, 2015.
1Current affiliation: Department of Laboratory Medicine, Karolinska Institute, 17177 Stockholm, Sweden. Address correspondence to Martine Daujat-Chavanieu, INSERM U1040, Institut de Recherche en BiothérapieHôpital St Eloi 80, Avenue Augustin Fliche, 34295 MontpellierCedex 5, France. Tel: +0033 4 67 33 06 23; Fax: +0033 4 67 33 04 59; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2557-2571, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686788
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Hepatic Stellate Cells Improve Engraftment of Human Primary Hepatocytes: A Preclinical Transplantation Study in an Animal Model

Ange-Clarisse Dusabineza,* Mustapha Najimi,† Noémi van Hul,* Vanessa Legry,* Dung Ngoc Khuu,† Leo A. van Grunsven,‡ Etienne Sokal,† and Isabelle A. Leclercq*

*Laboratory of Hepato-Gastroenterology, Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, UCL, Brussels, Belgium
†Laboratory of Pediatric Hepatology and Cell Therapy, Institut de Recherche Expérimentale et Clinique (IREC), Université catholique de Louvain, UCL, Brussels, Belgium
‡Liver Cell Biology Lab, Vrije Universiteit Brussel, VUB, Brussels, Belgium

Human hepatocytes are used for liver cell therapy, but the small number of engrafting cells limits the benefit of cell transplantation. We tested whether cotransplantation of hepatocytes with hepatic stellate cells (HSCs) could improve hepatocyte engraftment in vivo. Human primary hepatocytes were transplanted into SCID mice either alone or in a mixture with HSCs (quiescent or after culture activation) or LX-2 cells (ratio 20:1). Four weeks after transplantation into mouse livers, human albumin-positive (huAlb+) hepatocytes were found scattered. When cotransplanted in a mixture with HSCs or LX-2 cells, huAlb+
hepatocytes formed clusters and were more numerous occupying 2- to 5.9-fold more surface on the tissue section than in livers transplanted with hepatocytes alone. Increased huAlb mRNA expression in livers transplanted with the cell mixtures confirmed those results. The presence of HSCs increased the number of hepatocytes entrapped in the host liver at an early time point posttransplantation but not their proliferation in situ as assessed by cumulative incorporation of BrdU. Importantly, 4 weeks posttransplantation, we found no accumulation of aSMA+ -activated HSCs or collagen deposition. To follow the fate of transplanted HSCs, HSCs derived from GFP+ mice were injected into GFPlittermates: 17 h posttransplant, GFP+ HSCs were found in the sinusoids, without proliferating or actively producing ECM; they were undetectable at later time points. Coculture with HSCs improved the number of adherent hepatocytes, with best attachment obtained when hepatocytes were seeded in contact with activated HSCs. In vivo, cotransplantation of hepatocytes with HSCs into a healthy liver recipient does not generate fibrosis, but significantly improves the engraftment of hepatocytes, probably by ameliorating cell homing.

Key words: Cryopreserved hepatocytes; Hepatic stellate cells (HSCs); Transplantation; Engraftment; Extracellular matrix

Received December 27, 2013; final acceptance June 6, 2014. Online prepub date: January 20, 2015.
Address correspondence to Isabelle Leclercq, Prof., M.D., Ph.D., Laboratoire d’Hépato-Gastro-EntérologieInstitut de Recherche Expérimentale et Clinique, Université catholique de Louvain, GAEN53/79, Avenue Mounier, 53, B-1200 Brussels, Belgium. Tel: +32 2 764 52 73; Fax: +32 2 764 53 46; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2573-2584, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X686788
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Extensive Characterization of Platelet Gel Releasate From Cord Blood in Regenerative Medicine

Valentina Parazzi,* Cristiana Lavazza,* Valentina Boldrin,* Elisa Montelatici,* Francesco Pallotti,† Maurizio Marconi,‡ and Lorenza Lazzari*

*Cell Factory, Unit for Cellular Therapy and Cryobiology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy
†Unit of Anatomical Pathology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy
‡Blood Transfusion Center, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy

Platelet gel derived from peripheral blood is widely applied in many clinical fields of surgery as biomaterial containing growth factors with high proliferative properties. In 2010, we studied and patented a platelet gel derived from cord blood. In this study, due to the crucial role of the factors released by the platelet gel, we first extended the characterization of its releasate. Using a wide proteomic array and splitting the two components of the releasate, that is, platelets and plasma, we have been able to study their growth factor content. Interestingly, we discovered high levels of hormones and molecules able to support tissue growth in the cord blood platelet gel releasate and, in addition, higher concentrations of several angiogenic factors if compared with the peripheral blood counterpart. On the contrary, the latter was much richer in inflammatory factors. The second aim of our work was to study the effects on cell culture, immunophenotype, and function of mesenchymal stem cells exposed to these two platelet gel releasatesas substitute for the animal serum. Since our findings nicely show that the use of the peripheral versus the cord blood platelet gel releasate can differently influence the mesenchymal stem cell commitment, we can suggest that in addition to its peculiarangiogenic properties cord blood platelet gel releasate shows excellent proliferative properties as cell culture supplement.

Key words: Adipose mesenchymal stem cells; Human cord blood; Human peripheral blood; Platelet gel; Proteome array

Received August 27, 2014; final acceptance February 4, 2015. Online prepub date: February 18, 2015.
Address correspondence to Lorenza Lazzari, Ph.D., Cell Factory, Unit of Cellular 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. 24, pp. 2585-2599, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687452
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Umbilical Cord Mesenchymal Stem Cells Promote Breast Cancer Metastasis by Interleukin-8- and Interleukin-6-Dependent Induction of CD44+/CD24 Cells

Fengxia Ma,* Dandan Chen,‡ Fang Chen,* Ying Chi,* Zhibo Han,*† Xiaoming Feng,* Xue Li,* and Zhongchao Han*†

*State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Disease, Chinese Academy of Medical Sciences, Tianjin, China
†National Engineering Research Center of Cell Products/AmCellGene Co. Ltd., Tianjin, China
‡Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China

Although emerging evidence links mesenchymal stem cells (MSCs) with cancer metastasis, the underlying mechanisms are poorly understood. In the present study, we found that human umbilical cord-derived MSCs (UC-MSCs) promoted MCF-7 cell migration in vitro and metastasis in vivo. To explore the mechanisms, the characteristics of MCF-7 cells cocultured with UC-MSCs were assessed. The expression and secretion of interleukin-8 (IL-8) and IL-6 were induced in MCF-7 cells cocultured with UC-MSCs. However, neutralization of IL-8 or IL-6 secreted by UC-MSCs could attenuate the enhanced expression of IL-8 and IL-6 in MCF-7 cells cocultured with UC-MSCs, which subsequently alleviated the enhanced migration. Similar to UC-MSCs, exogenous human recombinant IL-8 or IL-6 also promoted IL-8 and IL-6 expression and MCF-7 cell migration. In addition to enhanced IL-8 and IL-6 expression, MCF-7 cells cocultured with UC-MSCs displayed enhanced mammosphere-forming ability and increased percentage of CD44+/CD24
cells. However, epithelial-to-mesenchymal transition (EMT) was not observed in MCF-7 cells cocultured with UC-MSCs. Taken together, these results suggested that IL-8 and IL-6 secreted by UC-MSCs activated the autocrine IL-8 and IL-6 signaling in MCF-7 cells and induced CD44+/CD24 cells, which subsequently promoted MCF-7 cell migration in vitro and metastasis in vivo.

Key words: Mesenchymal stem cells (MSCs); Interleukin-8 (IL-8); Interleukin-6 (IL-6); Epithelial-to-mesenchymal transition (EMT)

Received February 24, 2014; final acceptance January 29, 2015. Online prepub date: February 18, 2015.
Address correspondence to Zhongchao Han, State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Disease, Chinese Academy of Medical Sciences, 288 Nanjing Road, Tianjin 300020, China. Tel/Fax: +86-22-27210717; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it  or Fengxia Ma, State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Disease, Chinese Academy of Medical Sciences, 288 Nanjing Road, Tianjin 300020, China. Tel/Fax: +86-22-27210717; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2601-2614, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687570
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Amnion-Derived Mesenchymal Stem Cell Transplantation Ameliorates Dextran Sulfate Sodium-Induced Severe Colitis in Rats

Reizo Onishi,* Shunsuke Ohnishi,* Ryosuke Higashi,† Michiko Watari,‡ Kenichi Yamahara,§ Naoto Okubo,† Koji Nakagawa,† Takehiko Katsurada,* Goki Suda,* Mitsuteru Natsuizaka,* Hiroshi Takeda,† and Naoya Sakamoto*

*Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, Sapporo, Japan
†Laboratory of Pathophysiology and Therapeutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan
‡Department of Gynecology, Tenshi Hospital, Sapporo, Japan
§Department of Regenerative Medicine and Tissue Engineering, National Cerebral and Cardiovascular Center Research Institute, Osaka, Japan

Mesenchymal stem cells (MSCs) are a valuable cell source in regenerative medicine. Recently, several studies have shown that MSCs can be easily isolated from human amnion. In this study, we investigated the therapeutic effect of human amnion-derived MSCs (AMSCs) in rats with severe colitis. Colitis was induced by the administration of 8% dextran sulfate sodium (DSS) from day 0 to day 5, and AMSCs (1 × 106
cells) were transplanted intravenously on day 1. Rats were sacrificed on day 5, and the colon length and histological colitis score were evaluated. The extent of inflammation was evaluated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry. The effect of AMSCs on the inflammatory signals was investigated in vitro. AMSC transplantation significantly ameliorated the disease activity index score, weight loss, colon shortening, and the histological colitis score. mRNA expression levels of proinflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and migration inhibitory factor (MIF) were significantly decreased in the rectums of AMSC-treated rats. In addition, the infiltration of monocytes/macrophages was significantly decreased in AMSC-treated rats. In vitro experiments demonstrated that activation of proinflammatory signals induced by TNF-α or lipopolysaccharide (LPS) in immortalized murine macrophage cells (RAW264.7) was significantly attenuated by coculturing with AMSCs or by culturing with a conditioned medium obtained from AMSCs. Although the phosphorylation of IκB induced by TNF-α or LPS was not inhibited by the conditioned medium, nuclear translocation of NF-κB was significantly inhibited by the conditioned medium. Taken together, AMSC transplantation provided significant improvement in rats with severe colitis, possibly through the inhibition of monocyte/macrophage activity and through inhibition of NF-κB activation. AMSCs could be considered as a new cell source for the treatment of severe colitis.

Key words: Mesenchymal stem cells (MSCs); Amnion; Colitis; Macrophages; NF-κB

Received January 31, 2015; final acceptance February 22, 2015. Online prepub date: March 25, 2015.
Address correspondence to Shunsuke Ohnishi, M.D., Ph.D., Department of Gastroenterology and Hepatology, Hokkaido University Graduate School of Medicine, N15, W7, Kita-ku, Sapporo 060-8638, Japan. Tel: +81-11-716-1161; Fax: +81-11-706-7867; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2615-2627, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687543
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Immunoregulatory Effects of Mesenchymal Stem Cell-Derived Extracellular Vesicles on T Lymphocytes

Andrea Del Fattore,* Rosa Luciano,* Luisa Pascucci,† Bianca Maria Goffredo,‡ Ezio Giorda,§ Margherita Scapaticci,¶ Alessandra Fierabracci,# and Maurizio Muraca**

*Regenerative Medicine, Bambino Gesu Children’s Hospital, Rome, Italy
†Department of Veterinary Medicine, University of Perugia, Perugia, Italy
‡Department of Laboratory, Bambino Gesu Children’s Hospital, Rome, Italy
§Immunology Unit, Bambino Gesu Children’s Hospital, Rome, Italy
¶Laboratory Medicine, Figlie di San Camillo Hospital, Treviso, Italy
#Autoimmunity Laboratory, Immunology and Pharmacotherapy Area, Bambino Gesu Children’s Hospital, Rome, Italy
**Department of Women and Children’s Health, University of PadovaPadova, Italy

The immunomodulatory activity of mesenchymal stem cells (MSCs) is largely mediated by paracrine factors. We have recently shown that the immunosuppressive effects of MSCs on B lymphocytes in peripheral blood mononuclear cell (PBMC) culture can be reproduced by extracellular vesicles (EVs) isolated from MSC culture supernatants. Here we investigated the effect of bone marrow-derived MSC-EVs on T cells on PBMC cultures stimulated with anti-CD3/CD28 beads. Stimulation increased the number of proliferating CD3+
cells as well as of regulatory T cells (Tregs). Coculture with MSCs inhibited the proliferation of CD3+ cells, with no significant changes in apoptosis. Addition of MSC-EVs to PBMCs did not affect proliferation of CD3+ cells, but induced the apoptosis of CD3+ cells and of the CD4+ subpopulation and increased the proliferation and the apoptosis of Tregs. Moreover, MSC-EV treatment increased the Treg/Teff ratio and the immunosuppressive cytokine IL-10 concentration in culture medium. The activity of indoleamine 2,3-dioxygenase (IDO), an established mediator of MSC immunosuppressive effects, was increased in supernatants of PBMCs cocultured with MSCs, but was not affected by the presence of MSC-EVs. MSC-EVs demonstrate immunomodulatory effects on T cells in vitro. However, these effects and the underlying mechanisms appear to be different from those exhibited by their cells of origin.

Key words: Extracellular vesicles (EVs); Mesenchymal stem/stromal cells (MSCs); Regulatory T cells; Immunomodulation

Received July 15, 2014; final acceptance February 12, 2015. Online prepub date: February 18, 2015.
Address correspondence to Maurizio Muraca, M.D., Ph.D., Department of Women and Children’s Health, University of Padova, Via Giustiniani 3, 35128 Padova, Italy. Tel: +39 0498213574; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2629-2642, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687499
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
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Therapeutic Effects of Bone Marrow-Derived Mesenchymal Stem Cells in Models of Pulmonary and Extrapulmonary Acute Lung Injury

Ling Liu,1 Hongli He,1 Airan Liu, Jingyuan Xu, Jibin Han, Qihong Chen, Shuling Hu, Xiuping Xu, Yingzi Huang, Fengmei Guo, Yi Yang, and Haibo Qiu

Department of Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, Jiangsu, P. R. China

Bone marrow-derived mesenchymal stem cells (MSCs) offer a promising therapy for acute lung injury (ALI). However, whether the same MSC treatments possess similar potential for different ALI models is not fully clear. The present study evaluated the distribution and therapeutic effects of intravenous MSC administration for the treatment of intratracheal lipopolysaccharide (LPS)-induced intrapulmonary ALI and intravenous LPS/zymosan-induced extrapulmonary ALI, matched with lung injury severity, at 30 min and 1, 3, and 7 days. We found that MSC transplantation attenuated lung injury and inhibited lung inflammation in both ALI models. The benefits of MSCs were more significant in the intrapulmonary ALI mice. In vivo and ex vivo fluorescence imaging showed that MSCs primarily homed into the lung. However, more MSCs were recruited into the lungs of the intrapulmonary ALI mice than those of the extrapulmonary ALI mice over the time course. A few MSCs were also detected in the liver and spleen at days 3 and 7. In addition, the two ALI models showed different extrapulmonary organ dysfunction. A lower percentage of cell apoptosis and SDF-1α levels was found in the liver and spleen of the intrapulmonary ALI mice than in those of theextrapulmonary ALI mice. These results suggested that the two ALI models were accompanied with different degrees of extrapulmonary organ damage, which resulted in differences in the trafficking and accumulation of MSCs to the injured lung and consequently accounted for different therapeutic effects of MSCs for lung repair in the two ALI models. These data suggest that intravenous administration of MSCs has a greater potential for the treatment of intrapulmonary ALI than extrapulmonary ALI matched with lung injury severity; these differences were due to more recruitment of MSCs in the lungs of intrapulmonary ALI mice than those of extrapulmonary ALI mice. This finding may contribute to the clinical use of MSCs for the treatment of ALI.

Key words: Mesenchymal stem cells (MSCs); Acute lung injury (ALI); Lung repair; Homing; Retention

Received July 28, 2014; final acceptance February 7, 2015. Online prepub date: February 18, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Haibo Qiu, Department of Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast University, No. 87 Ding Jiaqiao, Nanjing (210009), Jiangsu, P. R. China. Tel: +86-25-83262551; 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. 24, pp. 2643-2655, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687219
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Systemic and Local Administration of Allogeneic Bone Marrow-Derived Mesenchymal Stem Cells Promotes Fracture Healing in Rats

Shuo Huang,*†‡1 Liangliang Xu,*†‡1 Yifeng Zhang,*† Yuxin Sun,*†‡ and Gang Li*†‡§

*Department of Orthopaedics and Traumatology, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, PR China
Lui Che Woo Institute of Innovative Medicine, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, PR China
‡The Chinese University of Hong Kong Shenzhen Research Institute, Shenzhen, PR China
§Key Laboratory for Regenerative Medicine, Ministry of Education, School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong SAR, China

Mesenchymal stem cells (MSCs) are immune privileged and a cell source for tissue repair. Previous studies showed that there is systemic mobilization of osteoblastic precursors to the fracture site. We hypothesized that both systemic and local administration of allogeneic MSCs may promote fracture healing. Bone marrow-derived MSCs and skin fibroblasts were isolated from GFP Sprague–Dawley rats, cultured, and characterized. Closed transverse femoral fracture with internal fixation was established in 48 adult male Sprague–Dawley rats, which were randomly assigned into four groups receiving PBS injection, MSC systemic injection, fibroblast systemic injection, and MSC fracture site injection; 2 × 106
cells were injected at 4 days after fracture. All animals were sacrificed at 5 weeks after fracture; examinations included weekly radiograph, micro-CT, mechanical testing, histology, immunohistochemistry, and double immunofluorescence. The callus size of MSC injection groups was significantly larger among all the groups. Radiographs and 3D reconstruction images showed that the fracture gaps united in the MSC injected groups, while gaps were still seen in the fibroblast and PBS injection groups. The mechanical properties were significantly higher in the MSC injection groups than those in the fibroblast and PBS groups, but no difference was found between the MSC local and systemic injection groups. Immunohistochemistry and double immunofluorescence demonstrated that GFP-positive MSCs were present in the callus in the MSC injection groups at 5 weeks after fracture, and some differentiated into osteoblasts. Quantitative analysis revealed the number of GFP-positive cells in the callus in the MSC systemic injection group was significantly lower than that of the MSC local injection group. The proportion of GFP osteoblasts in GFP-positive cells in the MSC systemic injection group was significantly lower than that of the MSC local injection group. These findings provide critical insight for developing MSC-based therapies, and systemic injection of allogeneic MSCs may be a novel treatment method for promoting fracture repair.

Key words: Allogeneic mesenchymal stem cells (MSCs); Systemic injection; Local injection; Fracture healing

Received November 13, 2014; final acceptance January 23, 2015. Online prepub date: February 2, 2015.
1These authors provided equal contribution to this work.
Address correspondence to Gang Li, M.B.B.S., D. Phil. (Oxon), Li Ka Shing Institute of Health Institute, Prince of Wales Hospital, The Chinese University of Hong Kong, Room 904, 9/F, Shatin, Hong Kong, SAR, PR China. Tel: (+852) 3763 6153; Fax: (+852) 2646 3020; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 24, pp. 2657-2666, 2015
0963-6897/15 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368915X687534
E-ISSN 1555-3892
Copyright © 2015 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Medium Attenuate Fibrosis in an Irreversible Model of Unilateral Ureteral Obstruction

Andrei F. da Silva, Kleiton Silva, Luciana A. Reis, Vicente P. C. Teixeira, and Nestor Schor

Nephrology Division, Department of Medicine, UNIFESP/EPM, São Paulo, Brazil

The therapeutic potential of mesenchymal stem cells (MSCs) and their conditioned medium (MSC-CM) has been extensively studied. MSCs can repair tissue, reduce local inflammation, and modulate the immune response. Persistent renal tubular interstitial inflammation results in fibrosis and leads to chronic kidney disease (CKD). Unilateral ureteral obstruction (UUO) is a very well-accepted renal fibrosis model. In this study, we evaluated factors influenced by the administration of MSCs or MSC-CM in the UUO model. MSCs extracted from rat bone marrow were cultivated in vitro and characterized by flow cytometry and cellular differentiation. Eight groups of female rats were used in experiments (n = 7, each), including Sham, UUO, UUO + MSC (obstruction + MSC), and UUO + CM (obstruction + MSC-CM) for 7 days of obstruction and Sham, UUO, UUO + MSC, and UUO + CM for 14 days of obstruction. The MSCs or MSC-CM was administered via the abdominal vena cava after total ligation of the left ureter. After 7 or 14 days, rats were euthanized, and serum and obstructed kidney samples were collected. MSCs or MSC-CM decreased the expression of molecules, such as Col1a1, α-SMA, and TNF-α. We also observed reductions in the levels of caspase 3, α-SMA, and PCNA in treated animals by immunohistochemistry. Our results suggest that the intravenous administration of MSCs or MSC-CM improves fibrosis progression and factors involved in apoptosis, inflammation, cell proliferation, and epithelial–mesenchymal transition in Wistar rats subjected to UUO, indicating a potential tool for preventing CKD.

Key words: Mesenchymal stem cells (MSCs); Conditioned medium (CM); Fibrosis; Unilateral ureteral obstruction (UUO); Repair

Received July 28, 2014; final acceptance February 11, 2015. Online prepub date: February 18, 2015.
Address correspondence to Nestor Schor, Renal Physiopathology and Stem Cell Research Laboratory, Rua Botucatu 740, 04023-900 São Paulo, Brazil. Tel: +55-11-5904 1699; Fax: +55-11-5904 1684; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it