Cell Transplantation 20(5) Abstracts

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Cell Transplantation, Vol. 20, pp. 593–607, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X532738
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Review

Cell Transplantation for Articular Cartilage Defects: Principles of Past, Present, and Future Practice

Yang Zi Jiang,* Shu Fang Zhang,* Yi Ying Qi,† Lin Lin Wang,* and Hong Wei Ouyang*‡

*Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, Hangzhou, China
†Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China
‡Department of Orthopaedic Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, China

As articular cartilage has very limited self-repair capability, the repair and regeneration of damaged cartilage is a major challenge. This review aims to outline the past, present, and future of cell therapies for articular cartilage defect repair. Autologous chondrocyte implantation (ACI) has been used clinically for more than 20 years, and the short, medium, and long-term clinical outcomes of three generation of ACI are extensively overviewed. Also, strategies of clinical outcome evaluation, ACI limitations, and the comparison of ACI clinical outcomes with those of other surgical techniques are discussed. Moreover, mesenchymal stem cells and pluripotent stem cells for cartilage regeneration in vitro, in vivo, and in a few clinical studies are reviewed. This review not only comprehensively analyzes the ACI clinical data but also considers the findings from state-of-the-art stem cell research on cartilage repair from bench and bedside. The conclusion provides clues for the future development of strategies for cartilage regeneration.

Key words: Cell transplantation; Cartilage repair; Chondrocytes; Stem cells

Address correspondence to Hong Wei Ouyang, Center for Stem Cell and Tissue Engineering, School of Medicine, Zhejiang University, 388 Yu Hang Tang Road, 310058 Hangzhou, China. Tel/Fax: 0086-0571-88208262; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 609–618, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536491
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Initial Observations of Cell-Mediated Drug Delivery to the Deep Lung

Arun Kumar,* Mark Glam,* Nagwa El-Badri,† Shyam Mohapatra,* Edward Haller,‡ Seungjoo Park,‡ Leslie Patrick,‡ Leigh Nattkemper,* Dawn Vo,* and Don F. Cameron‡

*Department of Internal Medicine, University of South Florida College of Medicine, Tampa, FL, USA
†Department of Obstetrics & Gynecology, University of South Florida College of Medicine, Tampa, FL, USA
‡Department of Pathology & Cell Biology, University of South Florida College of Medicine, Tampa, FL, USA

Using current methodologies, drug delivery to small airways, terminal bronchioles, and alveoli (deep lung) is inefficient, especially to the lower lungs. Urgent lung pathologies such as acute respiratory distress syndrome (ARDS) and post-lung transplantation complications are difficult to treat, in part due to the methodological limitations in targeting the deep lung with high efficiency drug distribution to the site of pathology. To overcome drug delivery limitations inhibiting the optimization of deep lung therapy, isolated rat Sertoli cells preloaded with chitosan nanoparticles were use to obtain a high-density distribution and concentration (92%) of the nanoparticles in the lungs of mice by way of the peripheral venous vasculature rather than the more commonly used pulmonary route. Additionally, Sertoli cells were preloaded with chitosan nanoparticles coupled with the anti-inflammatory compound curcumin and then injected intravenously into control or experimental mice with deep lung inflammation. By 24 h postinjection, most of the curcumin load (~90%) delivered in the injected Sertoli cells was present and distributed throughout the lungs, including the perialveloar sac area in the lower lungs. This was based on the high-density, positive quantification of both nanoparticles and curcumin in the lungs. There was a marked positive therapeutic effect achieved 24 h following curcumin treatment delivered by this Sertoli cell nanoparticle protocol (SNAP). Results identify a novel and efficient protocol for targeted delivery of drugs to the deep lung mediated by extratesticular Sertoli cells. Utilization of SNAP delivery may optimize drug therapy for conditions such as ARDS, status asthmaticus, pulmonary hypertension, lung cancer, and complications following lung transplantation where the use of high concentrations of anti-inflammatory drugs is desirable, but often limited by risks of systemic drug toxicity.

Key words: Sertoli cells; Drug delivery; Deep lung

Address correspondence to Don F. Cameron, Ph.D., Department of Pathology & Cell Biology, University of South Florida College of Medicine, MDC 11, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA. Tel:(813) 974-9434; Fax: (813) 974-2058; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 619–635, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536563
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Characterization and Functionality of Proliferative Human Sertoli Cells

Kitty Chui,*1 Alpa Trivedi,*†1 C. Yan Cheng,‡ Diana B. Cherbavaz,* Paul F. Dazin,* Ai Lam Thu Huynh,* James B. Mitchell,§ Gabriel A. Rabinovich,¶ Linda J. Noble-Haeusslein,† and Constance M. John*

*MandalMed, Inc., San Francisco, CA, USA
†Department of Neurosurgery, University of California, San Francisco, CA, USA
‡Population Council, New York, NY, USA
§Lonza Walkersville, Walkersville, MD, USA
¶Laboratory of Immunopathology, Institute of Biology and Experimental Medicine, CONICET, Buenos Aires, Argentina

It has long been thought that mammalian Sertoli cells are terminally differentiated and nondividing postpuberty. For most previous in vitro studies immature rodent testes have been the source of Sertoli cells and these have shown little proliferative ability when cultured. We have isolated and characterized Sertoli cells from human cadaveric testes from seven donors ranging from 12 to 36 years of age. The cells proliferated readily in vitro under the optimized conditions used with a doubling time of approximately 4 days. Nuclear 5-ethynyl-2´-deoxyuridine (EdU) incorporation confirmed that dividing cells represented the majority of the population. Classical Sertoli cell ultrastructural features, lipid droplet accumulation, and immunoexpression of GATA-4, Sox9, and the FSH receptor (FSHr) were observed by electron and fluorescence microscopy, respectively. Flow cytometry revealed the expression of GATA-4 and Sox9 by more than 99% of the cells, and abundant expression of a number of markers indicative of multipotent mesenchymal cells. Low detection of endogenous alkaline phosphatase activity after passaging showed that few peritubular myoid cells were present. GATA-4 and SOX9 expression were confirmed by reverse transcription polymerase chain reaction (RT-PCR), along with expression of stem cell factor (SCF), glial cell line-derived neurotrophic factor (GDNF), and bone morphogenic protein 4 (BMP4). Tight junctions were formed by Sertoli cells plated on transwell inserts coated with fibronectin as revealed by increased transepithelial electrical resistance (TER) and polarized secretion of the immunoregulatory protein, galectin-1. These primary Sertoli cell populations could be expanded dramatically in vitro and could be cryopreserved. The results show that functional human Sertoli cells can be propagated in vitro from testicular cells isolated from adult testis. The proliferative human Sertoli cells should have important applications in studying infertility, reproductive toxicology, testicular cancer, and spermatogenesis, and due to their unique biological properties potentially could be useful in cell therapy.

Key words: Sertoli cells; Blood–testis barrier (BTB); Sox9; Galectin-1

1These authors provided equal contribution to this work.
Address correspondence to Constance M. John, Ph.D., MandalMed, Inc., 665 3rd Street, Suite 250, San Francisco, CA 94107, USA. Tel: (415) 495-5570; Fax: (415) 495-5575; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 637–642, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536581
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Embryonic Stem Cell Transplantation Into Seminiferous Tubules: A Model for the Study of Invasive Germ Cell Tumors of the Testis

Unai Silván,1 Alejandro Díez-Torre, Ricardo Andrade, Jon Arluzea, Margarita Silió, and Juan Aréchaga

Laboratory of Stem Cells, Development and Cancer, Department of Cell Biology and Histology and Analytical and High Resolution Biomedical Microscopy Core Facility, University of the Basque Country, Vizcaya, Spain

Over the last 15 years, cell transplantation into seminiferous tubules has become a valuable tool to study germinal cell biology and related matters. This is particularly so, because the blood–testis permeability barrier establishes a sealed compartment which protect against certain influences such as immunological rejection. In the light of the functional and genetic similarities between carcinoma in situ (CIS) of the testis and embryonic stem (ES) cells, our laboratory has developed a tumor assay to study cancer invasion processes in testicular germ cell tumors (TGCT) based on the transplantation of ES cells into the seminiferous tubules. Here, we describe this new tumor assay and provide additional information regarding the transplantation techniques used and their application for the study of TGCTs. Finally, we discuss the practical implications of our experimental approach and its potential application for the understanding of TGCT invasive processes and the development of new antineoplastic strategies.

Key words: Seminiferous tubule microinjection; Embryonic stem cell transplantation; Embryonal carcinoma; Carcinoma in situ of testis; Testis teratocarcinoma pathogenesis

1Present address: Maurice E. Müller Institute for Structural Biology, Biozentrum, University of Basel, Basel, Switzerland.
Address correspondence to Prof. Juan Aréchaga, Department of Cell Biology and Histology, Faculty of Medicine and Dentistry, University of the Basque Country, E-48940 Leioa, Vizcaya, Spain. Tel: +34-94-601-2883; Fax: +34-94 601-2883; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 643–654, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536518
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Mesenchymal Stem Cells From Chorionic Villi and Amniotic Fluid Are Not Susceptible to Transformation After Extensive In Vitro Expansion

Antonella Poloni,* Giulia Maurizi,* Lucia Babini,* Federica Serrani,* Eleonora Berardinelli,* Stefania Mancini,* Benedetta Costantini,* Giancarlo Discepoli,† and Pietro Leoni*

*Clinica di Ematologia, Dipartimento Scienze Mediche e Chirurgiche, Università Politecnica delle Marche, Ancona, Italy
†Laboratorio di Citogenetica e Genetica Molecolare, Clinica di Pediatria, Ospedali Riuniti, Ancona, Italy

Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering. Increasing evidence suggests that MSCs isolated from fetal tissues are more plastic and grow faster than adult MSCs. In this study, we characterized human mesenchymal progenitor cells from chorionic villi (CV) and amniotic fluid (AF) isolated during the first and second trimesters, respectively, and compared them with adult bone marrow-derived MSCs (BM). We evaluated 10 CV, 10 AF, and 6 BM samples expanded until the MSCs reached senescence. We used discarded cells from prenatal analyses for all the experiments. To evaluate the replicative stability of these cells, we studied the telomerase activity, hTERT gene transcription, and telomere length in these cells. Spontaneous chromosomal alterations were excluded by cytogenetic analysis. We studied the expression of c-myc and p53, tumor-associated genes, at different passage in culture and the capacity of these cells to grow in an anchorage-independent manner by using soft agar assay. We isolated homogeneous populations of spindle-shaped CV, AF, and BM cells expressing mesenchymal immunophenotypic markers throughout the period of expansion. CV cells achieved 14 ± 0.9 logs of expansion in 118 days and AF cells achieved 21 ± 0.9 logs in 118 days, while BM cells achieved 11 × 0.4 logs in 84 days. Despite their high proliferation capacity, fetal MSCs showed no telomerase activity, no hTERT and c-myc transcriptions, and maintained long, stable telomeres. A constant expression level of p53 and a normal karyotype were preserved throughout long-term expansion, suggesting the safety of fetal MSCs. In conclusion, our results indicate that fetal MSCs could be an alternative, more accessible resource for cell therapy and regenerative medicine.

Key words: Fetal mesenchymal stem cells (MSCs); Telomere; Telomerase activity; Human telomerase reverse transcriptase (hTERT)

Address correspondence to Antonella Poloni, Clinica di Ematologia, Dipartimento Scienze Mediche e Chirurgiche, Università Politecnica delle Marche, Via Tronto 60020 Ancona, Italy. Tel: 0039 071 2206112 or 0039 071 2206108; Fax: 0039 071 2206108; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 655–667, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536473
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Immunogenicity and Immunomodulatory Properties of Umbilical Cord Lining Mesenchymal Stem Cells

Tobias Deuse,*†‡ Mandy Stubbendorff,† Karis Tang-Quan,†‡ Neil Phillips,§ Mark A. Kay,§ Thomas Eiermann,¶ Thang T. Phan,#** Hans-Dieter Volk,†† Hermann Reichenspurner,*† Robert C. Robbins,‡ and Sonja Schrepfer*†‡

*Cardiovascular Surgery, University Heart Center Hamburg, Hamburg, Germany
†TSI-lab, Cardiovascular Research Center, University Hospital Hamburg-Eppendorf, Hamburg, Germany
‡Cardiothoracic Surgery, Stanford University School of Medicine, Standord, CA, USA
§Pediatrics, Human Gene Therapy, Stanford University School of Medicine, Stanford, CA, USA
¶Transfusion Medicine, HLA-Lab, University Hospital Hamburg-Eppendorf, Hamburg, Germany
#Surgery, Yong Loo Lin School of Medicine, Singapore
*CellResearchCorp, Clinical Research Centre, Singapore
††BCRT, Charite, Campus Virchow-Klinikum, Berlin, Germany

We here present an immunologic head-to-head comparison between human umbilical cord lining mesenchymal stem cells (clMSCs) and adult bone marrow MSCs (bmMSCs) from patients >65 years of age. clMSCs had significantly lower HLA class I expression, higher production of tolerogenic TGF-β and IL-10, and showed significantly faster proliferation. In vitro activation of allogeneic lymphocytes and xenogeneic in vivo immune activation was significantly stronger with bmMSCs, whereas immune recognition of clMSCs was significantly weaker. Thus, bmMSCs were more quickly rejected in immunocompetent mice. IFN-γ at 25 ng/ml increased both immunogenicity by upregulation of HLA class I/ HLA-DR expression and tolerogenicity by increasing intracellular HLA-G and surface HLA-E expression, augmenting TGF-β and IL-10 release, and inducing indoleamine 2,3-dioxygenase (IDO) expression. Higher concentrations of IFN-γ (>50 ng/ml) further enhanced the immunosuppressive phenotype of clMSCs, more strongly downregulating HLADR expression and further increasing IDO production (at 500 ng/ml). The net functional immunosuppressive efficacy of MSCs was tested in mixed lymphocyte cultures. Although both clMSCs and bmMSCs significantly reduced in vitro immune activation, clMSCs were significantly more effective than bmMSCs. The veto function of both MSC lines was enhanced in escalating IFN-γ environments. In conclusion, clMSCs show a more beneficial immunogeneic profile and stronger overall immunosuppressive potential than aged bmMSCs.

Key words: Mesenchymal stem cells (MSCs); Immunogenicity; Immunomodulation; Stem cell transplantation

Address correspondence to Sonja Schrepfer, M.D., Ph.D., Heisenbergprofessor at the University Heart Center Hamburg, Transplant and Stem Cell Immunobiology Lab (TSI), Martinistr. 52, Campus Research, N27, Room 068, 20146 Hamburg, Germany. Tel: +49-40-7410-5 9982; Fax: +49-40-7410-5 9663; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 669–680, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536509
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Augmenting Therapy of Ovarian Cancer Efficacy by Secreting IL-21 Human Umbilical Cord Blood Stem Cells in Nude Mice

Weihua Hu,*1 Jing Wang,†1 Jun Dou,*1 Xiangfeng He,* Fengshu Zhao,* Cuilian Jiang,* Fangliu Yu,* Kai Hu,* Lili Chu,‡ Xiaoli Li,* and Ning Gu§

*Department of Pathogenic Biology and Immunology, Medical School, Southeast University, Nanjing, China
†Department of Gynecology & Obstetrics, Zhongda Hospital, Southeast University, Nanjing, China
‡Paediatric Research Institute, Nanjing Children’s Hospital, Nanjing, China
§School of Biological Science & Medical Engineering, Southeast University, Nanjing, China

In the present study, CD34+ human umbilical cord blood stem cells (UCBSCs) were engineered to express interleukin-21 (IL-21) and then were transplanted into A2780 ovarian cancer xenograft-bearing Balb/c nude mice. The therapeutic efficacy of this procedure on ovarian cancer was evaluated. The findings from the study indicated that UCBSCs did not form gross or histological teratomas until up to 70 days postinjection. The CD34+ UCBSC-IL-21 therapy showed a consistent effect in the ovarian cancer of the treated mice, delaying the tumor appearance, reducing the tumor sizes, and extending life expectancy. The efficacy was attributable to keeping CD34+ UCBSC-IL-21 in the neoplastic tissues for more than 21 days. The secreted IL-21 not only increased the quantity of CD11a+ and CD56+ NK cells but also increased NK cell cytotoxicities to YAC-1 cells and A2780 cells, respectively. The efficacy was also associated with enhancing the levels of IFN-γ, IL-4, and TNF-α in the mice as well as the high expressions of the NKG2D and MIC A/B molecules in the tumor tissues. This study suggested that transferring CD34+ UCBSC-IL-21 into the nude mice was safe and feasible in ovarian cancer therapy, and that the method would be a promising new strategy for clinical treatment of ovarian cancer.

Key words: Interleukin-21; Umbilical cord blood stem cells (UCBSCs); Ovarian cancer; Gene therapy

1These authors provided equal contribution to this work.
Address correspondence to Dr. Jun Dou, Department of Pathogenic Biology and Immunology, Medical College, Southeast University, Nanjing 210009, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Dr. Ning Gu, School of Biological Science & Medical Engineering, Southeast University, Nanjing 210096, China. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 681–691, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536545
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Bone Marrow Stromal Cells and Skin Fibroblasts Inhibit Natural Killer Cell Proliferation and Cytotoxic Activity

Amandine Pradier,* Jakob Passweg,* Jean Villard,† and Vincent Kindler*

*Hematology Service, Geneva University Hospitals and Faculty of Medicine, University of Geneva, Geneva, Switzerland
†Transplantation Immunology Unit, Geneva University Hospitals and Faculty of Medicine, University of Geneva, Geneva, Switzerland

Mesenchymal stromal cells (MSCs) are potent immunomodulators that have successfully been used to circumvent various types of inflammations, including steroid-resistant graft-versus-host disease. Although initially believed to be restricted to multipotent MSCs, this immunoregulatory function is shared with differentiated cells from the mesenchymal lineage such as skin fibroblasts (SFs). Mesenchymal cell-induced immunoregulation is so potent that it may allow the reactivation of dormant malignancies, a fact that would preclude using such cells as therapeutic agents. Because NK cells are pivotal effectors controlling tumor cell containment we investigated the effect of allogenic MSCs and SFs on NK cell function in vitro. When NK cells were incubated with IL-15 and MSCs or SFs for 6 days, their proliferation and cytotoxic activity were significantly decreased compared to NK cells cultured with IL-15 alone or with human venous endothelial cells. Cytotoxic activity inhibition reached 86% when assayed on MHC-I+ allogenic primary hematopoietic blasts, and was associated with a significant decrease in cytolytic granule exocytosis and in perforin release. Stromal cell-mediated inhibition was effective only if cell–cell proximity was long lasting: when NK cells were activated with IL-15 in the absence of MSCs and assayed for cytotoxicity in their presence no inhibition occurred. MSC inhibition was ultimately mediated by a soluble factor generated upon incubation with NK cells activated by IL-15 or IL-2. The indoleamine 2,3 dioxygenase was activated in MSCs and SFs because L-kynurenine was detected in inhibitory supernatants, but its blockade did not restore NK cell functions. The profound inhibition of cytotoxic activity directed against allogenic hematopoietic blasts exerted by MSCs and SFs on NK cells may be a concern. Should this occur in vivo it may induce the inability of NK cells to control residual or dormant malignant diseases after infusion of therapeutic MSCs.

Key words: Mesenchymal stromal cells (MSCs); Skin fibroblast; Graft-versus-host disease (GVHD); Graft-versus-leukemia (GVL); Immunoregulation; Soluble inhibitor

Address correspondence to Vincent Kindler, Ph.D., P.D., Hematology Service, Geneva University Hospital, 4, Gabrielle-Perret-Gentil,1211 Geneva 14, Switzerland. Tel: +41.22.372.39.42; Fax: +41.22.372.98.55; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 693–706, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X550198
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Enhancement of Wound Healing by Human Multipotent Stromal Cell Conditioned Medium: The Paracrine Factors and p38 MAPK Activation

Tu-Lai Yew,*1 Yeh-Ting Hung,†‡1 Hsin-Yang Li,§¶1 Hsin-Wei Chen,# Ling-Lan Chen,# Kuo-Shu Tsai,# Shih-Hwa Chiou,#** Kuan-Chong Chao,§¶ Tung-Fu Huang,†† Hen-Li Chen,* and Shih-Chieh Hung†#**††

*Institute of Oral Biology, Department of Dentistry, National Yang-Ming University, Taipei, Taiwan
†Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
‡Buddhist Dalin Tzuchi General Hospital, Dalin, Taiwan
§Division of Obstetrics and Gynecology, Faculty of Medicine, National Yang-Ming University, Taipei, Taiwan
¶Obstetrics and Gynecology, Taipei Veterans General Hospital, Taipei, Taiwan
#Stem Cell Laboratory, Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan
**Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan
††Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan

Wound healing can be improved by transplanting mesenchymal stem cells (MSCs). In this study, we have demonstrated the benefits of the conditioned medium derived from human MSCs (CM-MSC) in wound healing using an excisional wound model. CM-MSC accelerated wound closure with increased reepithelialization, cell infiltration, granulation formation, and angiogenesis. Notably, CM-MSC enhanced epithelial and endothelial cell migration, suggesting the contribution of increased cell migration to wound healing enhanced by CM-MSC. Cytokine array, ELISA analysis, and quantitative RT-PCR revealed high levels of IL-6 in CM-MSC. Moreover, IL-6 added to the preconditioned medium enhanced both cell migration and wound healing, and antibodies against IL-6 blocked the increase in cell motility and wound closure by CM-MSC. The IL-6 secretory pathway of MSCs was inhibited by SB203580, an inhibitor of p38 MAPK or siRNA against p38 MAPK, suggesting IL-6 secretion by MSCs is mediated through the activation of p38 MAPK. Inactivation of p38 MAPK also reduced the expression and production of IL-8 and CXCL1 by MSCs, both of which were also demonstrated to enhance cell migration and wound closure. Thus, our data suggest MSCs promote wound healing through releasing a repertoire of paracrine factors via activation of p38 MAPK, and the CM-MSC may be applied to enhance wound healing.

Key words: Wound healing; Mesenchymal stem cells (MSCs); Conditioned medium; Paracrine factors; p38 MAPK

1These authors provided equal contribution to this work.
Address correspondence to Shin-Chieh Hung, Department of Medical Research and Education, Veterans General Hospital-Taipei 201, Sec. 2, Shih-Pai Road, Taipei, 11217, Taiwan. Tel: +886-2-28757557, ext. 118; Fax: +886-2-28265164; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Hen-Li Chen, Institute of Oral Biology, Department of Dentistry, National Yang-Ming University, Taipei 112, Taiwan155, Sec. 2, Linong Street, Taipei, 11217, Taiwan. Tel: +886-2-28267314; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 707–726, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536590
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Differential Hematopoietic Supportive Potential and Gene Expression of Stroma Cell Lines From Midgestation Mouse Placenta and Adult Bone Marrow

Yingchun Wang,* Lubov Nathanson,† and Ian K. McNiece*

*Interdisciplinary Stem Cell Institute, University of Miami, Miami, FL, USA
†Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, FL, USA

During mouse embryogenesis, hematopoietic development takes place in several distinct anatomic locations. The microenvironment of different hematopoietic organs plays an important role in the proliferation and maturation of the hematopoietic cells. We hypothesized that fetal stromal cells would be distinct to adult bone marrow (BM)-derived stromal cells because the BM contributes mainly to the homeostasis of hematopoietic stem cells (HSCs), while extensive expansion of HSCs occurs during fetal development. Here we report the establishment of stromal cell lines from fetal hematopoietic organs, namely aorta-gonad-mesonephros (AGM), midgestation placenta (PL), and fetal liver (FL) together with adult bone marrow (BM). The growth patterns and hematopoietic supportive potential were studied. Their phenotypic and molecular gene expression profiles were also determined. Stromal cell lines from each tissue were able to support cobblestone area formation of BM c-Kit+Sca-1+ hematopoietic cells: 22 (22/47) from AGM, three (3/4) from PL, three (3/4) from FL, and three (3/3) from BM. There were similar levels of expansion of total mononuclear cells (TMNs) when HSCs were cocultured with fetal stroma and adult BM stroma. However, PL-derived stromal cells supported higher levels of generation of colony-forming progenitor cell (CFU-C), indicated by more colonies and colonies with significantly larger size. Flow cytometric analysis of the PL1 cells demonstrated a phenotype of CD45, CD105+, Sca-1+, CD34+, and CD49d+, compared to adult BM1 cells, which were CD45, CD105+, Sca-1+, CD34, and CD49d. Using Affymetrix microarray analysis, we identified that genes specifically express in endothelial cells, such as Tie1, Tek, Kdr, Flt4, Emcn, Pecam1, Icam2, Cdh5, Esam1, Prom1, Cd34, and Sele were highly expressed in stroma PL1, consistent with an endothelial phenotype, while BM1 expressed a mesenchymal stromal phenotype. In summary, these data demonstrate distinct characteristics of stromal cells that provide insights into the microenvironmental control of HSCs.

Key words: Hematopoietic stem cell; Stromal cells; Microenvironment; Transcriptional gene expression

Address correspondence to Ian K. McNiece, Interdisciplinary Stem Cell Institute, Leonard M. Miller School of Medicine, BRB Room 908, 1501 N.W. 10th Ave., Miami, FL 33136, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 727–739, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536554
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparative Study of Methods for Administering Neural Stem/Progenitor Cells to Treat Spinal Cord Injury in Mice

Yuichiro Takahashi,*† Osahiko Tsuji,* Gentaro Kumagai,‡ Chikako Miyauchi Hara,† Hirotaka James Okano,† Atsushi Miyawaki,§¶ Yoshiaki Toyama,* Hideyuki Okano,† and Masaya Nakamura*

*Department of Orthopaedic Surgery, School of Medicine, Keio University, Tokyo, Japan
†Department of Physiology, School of Medicine, Keio University, Tokyo, Japan
‡Department of Orthopaedic Surgery, School of Medicine, Hirosaki University, Aomori, Japan
§Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, Saitama, Japan
¶Life Function and Dynamics, ERATO, JST, Saitama, Japan

To investigate potential cures for spinal cord injury (SCI), several researchers have transplanted neural stem/ progenitor cells (NS/PCs) into the injured spinal cord by different procedures, including intralesional (IL), intrathecal (IT), and intravenous (IV) injection. However, there are no reports quantifying or comparing the number of cells successfully transplanted to the lesion site by each procedure in vivo. The purpose of the present study was to determine the optimal method of cell transplantation to the SCI site in terms of grafted cell survival and safety. For this purpose, we developed mouse NS/PCs that expressed a novel Venusluciferase fusion protein that enabled us to detect a minimum of 1,000 grafted cells in vivo by bioluminescence imaging (BLI). After inducing contusive SCI at the T10 level in mice, NS/PCs were transplanted into the injured animals three different ways: by IL, IT, or IV injection. Six weeks after the transplantation, BLI analysis showed that in the IL group, the luminescence intensity of the grafted cells had decreased to about 10% of its initial level, and appeared at the site of injury. In the IT group, the luminescence of the grafted cells, which was distributed throughout the entire subarachnoid space immediately after transplantation, was detected at the injured site 1 week later, and by 6 weeks had gradually decreased to about 0.3% of its initial level. In the IV group, no grafted cells were detected at the site of injury, but all of these mice showed luminescence in the bilateral chest, suggesting pulmonary embolism. In addition, one third of these mice died immediately after the IV injection. In terms of grafted cell survival and safety, we conclude that the IL application of NS/PCs is the most effective and feasible method for transplanting NS/PCs into the SCI site.

Key words: Spinal cord injury (SCI); Transplantation; Neural stem/progenitor cells (NS/PCs); Bioluminescence imaging (BLI)

Address correspondence to Masaya Nakamura, Department of Orthopaedic Surgery, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan. Tel: +81-3-3353-1211; Fax: +81-3353-6597; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Hideyuki Okano, Department of Physiology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan. Tel: +81-3-5363-3747; Fax: +81-3-3357-5445; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 741–751, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536536
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Labeling Stem Cells With a Near-Infrared Fluorescent Heptamethine Dye for Noninvasive Optical Tracking

Chao Zhang,*†1 Xu Tan,*1 Li Tan,* Tao Liu,* Dengqun Liu,* Lilong Zhang,* Song Fan,* Yongping Su,* Tianmin Cheng,* Yue Zhou,† and Chunmeng Shi*

*Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing, China
†Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Chongqing, China

Near-infrared (NIR) fluorescent agents hold great promise for noninvasive in vivo imaging. We have recently reported that a NIR fluorescent heptamethine dye, IR-780 iodide, exhibits unique optical properties for biomedical imaging. On the basis of this foregoing work, we further describe here the potential application of IR-780 iodide as a novel NIR agent for stem cell labeling and tracking. The labeling efficiency, subcellular localization, and the effects on cell viability and differentiation of IR-780 iodide were investigated. The in vivo distribution of stem cells after intravenous transplantation was traced by whole-body animal NIR imaging. Our results showed that IR-780 iodide exhibited superior labeling efficiency and biocompatibility with unique optical properties. Following whole-body NIR imaging, the pulmonary passage of stem cells was noninvasively visualized in rats after systemic transplantation of IR-780 iodide-labeled stem cells through intravenous delivery. With this NIR imaging method, we further confirmed that pretreatment with sodium nitroprusside (SNP), a vasodilator agent, significantly reduced the cell trapping in the lung and increased the cell passage through the lung capillaries. Our study suggests that IR-780 iodide may represent an effective NIR fluorophore for stem cell labeling and tracking.

Key words: Near-infrared (NIR) imaging; Heptamethine dye; IR-780 iodide; Stem cells

1These authors provided equal contribution to this work.
Address correspondence to Chunmeng Shi, Institute of Combined Injury, Third Military Medical University, 30 Gaotanyan Road, Chongqing, 400038, China. Tel: 86-23-68752280; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Yue Zhou, Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China.


Cell Transplantation, Vol. 20, pp. 753–766, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536572
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Construction of a Portal Implantable Functional Tissue-Engineered Liver Using Perfusion-Decellularized Matrix and Hepatocytes in Rats

Ji Bao,*1 Yujun Shi,*1 Huaiqiang Sun,* Xiangli Yin,† Ruina Yang,* Li Li,* Xi Chen,† and Hong Bu†

*Key Laboratory of Transplant Engineering and Immunology, West China Hospital, Sichuan University, Chengdu, Sichuan, China
†Department of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan, China

Innovative cell-based therapies, including hepatic tissue engineering following hepatocyte transplantation, are considered as theoretical alternatives to liver transplant or for partial replacement of liver function in patients. However, recent progress in hepatic tissue engineering has been hampered by low initial hepatocyte engraftment and insufficient blood supply in vivo. We developed an intact 3D scaffold of an extracellular matrix (ECM) derived from a decellularized liver lobe, with layer-by-layer (LbL) heparin deposition to avoid thrombosis, which we repopulated with hepatocytes and successfully implanted as a tissue-engineered liver (TEL) into the portal system. The TEL provided sufficient volume for transplantation of cell numbers representing up to 10% of whole-liver equivalents and was perfused by portal vein blood. Treatment of extended hepatectomized rats with a TEL improved liver function and prolonged survival; mean lifespan was extended from 16 to 72 h. At 72 h postoperation, the TEL sustained functional and viable hepatocytes. In conclusion, we propose the TEL as a state-of-the-art substitute for whole-liver transplantation and as a proof of concept for the technology that will eventually allow for the transplantation of a reconstituted liver.

Key words: Tissue-engineered liver; Hepatocyte; Extracellular matrix (ECM); Liver perfusion; Anticoagulant

1These authors provided equal contribution to this work.
Address correspondence to Hong Bu, M.D., Ph.D., 37# Guoxue Road, Chengdu, Sichuan, China, 610041. Tel: 86-28-85164030; Fax: 86-28-85164034; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 767–774, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536608
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Novel Cell Culture Model Using Pure Hydrostatic Pressure and a Semipermeable Membrane Pouch

Shuichi Mizuno

Department of Orthopedic Surgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

Cell constructs and culture methods are essential tools in tissue engineering. The cell construct should be equivalent to the native cartilage it is intended to replace. Thus, three-dimensional cell constructs are usually composed of a high density of cells and dense extracellular matrix. However, dense constructs suffer from a lack of passive nutrient supply, gas exchange, and removal of degraded debris. We have developed a novel hydrostatic pressure/perfusion culture system that improves the quality of neo-tissues, providing an automated and affordable system for clinical applications. We have also developed a semipermeable membrane pouch that contains a fragile amorphous cell carrier. Although amorphous material is difficult to handle, it is a useful medium in which to deliver cells to the desired site via injection. We evaluated phenotypes of bovine articular chondrocytes embedded in a collagen type I gel enclosed within membrane pouches permeable to molecules of various sizes. Constant or cyclic hydrostatic pressure was externally applied to the medium phase with a new culture system. Accumulation of cartilage specific matrix was promoted with a 500-kDa cutoff membrane pouch and cyclic hydrostatic pressure at 0.5 MPa, 0.5 Hz. This new method will be useful in the delivery of engineered cells to a desired tissue in regenerative medicine.

Key words: Tissue engineering; Hydrostatic pressure; Semipermeable membrane pouch; Cell culture method; Chondrocytes

Address correspondence to Shuichi Mizuno, Ph.D., Orthopedic Research, Brigham and Women’s Hospital, 75 Francis St., Boston, MA 02115, USA. Tel: 617-732-6335; Fax: 617-732-6705; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 775–781, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X539056
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Pathogen Inactivation of Human Serum Facilitates its Clinical Use for Islet Cell Culture and Subsequent Transplantation

Magnus U. Ståhle, Daniel Brandhorst, Olle Korsgren, and Folke Knutson

Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden

Serum is regarded as an essential supplement to promote survival and growth of cells during culture. However, the potential risk of transmitting diseases disqualifies the use of serum for clinical cell therapy in most countries. Hence, most clinical cell therapy programs have replaced human serum with human serum albumin, which can result in inferior quality of released cell products. Photochemical treatment of different blood products utilizing Intercept® technology has been shown to inactivate a broad variety of pathogens of RNA and DNA origin. The present study assesses the feasibility of using pathogen-inactivated, blood group-compatible serum for use in human pancreatic islet culture. Isolated human islets were cultured at 37°C for 3–4 days in CMRL 1066 supplemented with 10% of either pathogen-inactivated or nontreated human serum. Islet quality assessment included glucose-stimulated insulin release (perifusion), ADP/ATP ratio, cytokine expression, and posttransplant function in diabetic nude mice. No differences were found between islets cultured in pathogen-inactivated or control serum regarding stimulated insulin release, intracellular insulin content, and ADP/ATP ratio. Whether media was supplemented with treated or nontreated serum, islet expression of IL-6, IL-8, MCP-1, or tissue factor was not affected. The final diabetes-reversal rate of mice receiving islets cultured in pathogen-inactivated or nontreated serum was 78% and 87%, respectively (NS). As reported here, pathogen-inactivated human serum does not affect viability or functional integrity of cultured human islets. The implementation of this technology for RNA- and DNA-based pathogen inactivation should enable reintroduction of human serum for clinical cell therapy.

Key words: Cell therapy; Cell transplantation; Cell culture; Serum; Pathogen inactivation

Address correspondence to Magnus Ståhle, Department of Immunology, Genetics and Pathology, Uppsala University, Rudbeck Laboratory (C11), Dag Hammarskjölds väg 20, S-75185 Uppsala, Sweden. Tel: +46-70-4250507; Fax: +46-18-507866; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 783–788, 2011
0963-6897/11 $90.00 + .00
DOI: 10.3727/096368910X536527
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

High Vascular Density and Oxygenation of Pancreatic Islets Transplanted in Clusters Into Striated Muscle

Johanna Svensson,*1 Joey Lau,*1 Monica Sandberg,* and Per-Ola Carlsson*†

*Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden
†Department of Medical Sciences, Uppsala University, Uppsala, Sweden

Pancreatic islet transplantation is presently almost exclusively performed using the intraportal route for transplantation into the liver. However, islets at this site are poorly revascularized and, when also considering the poor long-term results of clinical islet transplantation, there has in recent years emerged an increased interest to evaluate alternative sites for islet transplantation. Striated muscle is easily accessible and has for decades been used for autotransplantation of parathyroid glands. Moreover, it is almost the only tissue in the adult where physiological angiogenesis occurs. The present study tested the hypothesis that striated muscle would provide good conditions for revascularization and oxygenation of transplanted islets. Because we previously have observed similar revascularization of islets implanted to the renal subcapsular site and intraportally into the liver, islets grafted to the kidney were for simplicity besides native islets used for comparison. Islets grafted into muscle were found to have three times more blood vessels than corresponding islets at the renal subcapsular site at 2 month follow-up, but still less vascular numbers than native islets. The oxygen tension in 2-month-old intramuscular islet grafts was sixfold higher than in corresponding renal subcapsular grafts, and 70% of that in native islets. However, the oxygenation of surrounding muscle was only 50% of that in renal cortex, and connective tissue constituted a larger proportion of the intramuscular than the renal subcapsular grafts, suggesting exaggerated early islet cell death at the former site. We conclude that the intramuscular site provides excellent conditions for vascular engraftment, but that interventions to improve early islet survival likely are needed before clinical application. Such could include bioengineered matrices that not only spatially disperse the islet, but also could provide local supply of oxygen carriers, growth and survival factors, strategies that are much more easily applied at the intramuscular than the intrahepatic site.

Key words: Engraftment; Oxygenation; Islet graft; Angiogenesis; Muscle

1These authors provided equal contribution as first authors.
Address correspondence to Per-Ola Carlsson, Department of Medical Cell Biology, Uppsala University, Husargatan 3, Box 571, SE-751 23 Uppsala, Sweden. Tel: +46 18 4714425; Fax: +46 18 4714059; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it