Cell Transplantation 20(11-12) Abstracts

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Cell Transplantation, Vol. 20, pp. 1659–1672, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X564985
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Functional Modulation of Choroid Plexus Epithelial Clusters In Vitro for Tissue Repair Applications

C. G. Thanos,*† B. E. Bintz,*† Moses Goddard,* K. Boekelheide,† Susan Hall,† and D. F. Emerich‡

*CytoSolv, Inc., Providence, RI, USA
†Department of Pathology and Laboratory Medicine, Brown University, Providence, RI, USA
‡Glocester Institute of Regenerative Medicine, Glocester, RI, USA

One of the primary obstacles in the restoration or repair of damaged tissues is the temporospatial orchestration of biological and physiological events. Cellular transplantation is an important component of tissue repair as grafted cells can serve as replacement cells or as a source of secreted factors. But few, if any, primary cells can perform more than a single tissue repair function. Epithelial cells, derived from the choroid plexus (CP), are an exception to this rule, as transplanted CP is protective and regenerative in animal models as diverse as CNS degeneration and dermal wound repair. They secrete a myriad of proteins with therapeutic potential as well as matrix and adhesion factors, and contain responsive cytoskeletal components potentially capable of precise manipulation of cellular and extracellular niches. Here we isolated CP from neonatal porcine lateral ventricles and cultured the cells under a variety of conditions to specifically modulate tissue morphology (2D vs. 3D) and protein expression. Using qRT-PCR analysis, transmission electron microscopy, and gene microarray studies we demonstrate a fine level of control over CP epithelial cell clusters opening further opportunities for exploration of the therapeutic potential of this unique tissue source.

Key words: Choroid plexus (CP); Spheroid; Growth factors; 3D culture; Vascular endothelial growth factor (VEGF); Neuroprotection

Address correspondence to Christopher G. Thanos, Ph.D., CytoSolv, Inc., 117 Chapman St., Suite 107, Providence, RI 02905, USA. Tel: 401-573-2001; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1673–1691, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X576009
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Characterization of Olfactory Stem Cells

Andrew Wetzig,1 Alan Mackay-Sim, and Wayne Murrell

Eskitis Institute for Cell and Molecular Therapies, National Centre for Adult Stem Cell Research, Griffith University, Nathan, Queensland, Australia

There is worldwide enthusiasm for the prospect of some kind of cellular transplant therapy for repair of failing organs. The olfactory mucosa of a patient’s nose is easily biopsied to provide a ready source of multipotent cells. In this article we address practical issues pertinent to using olfactory neural stem cells for tissue repair. These cells are emerging as potentially most significant candidates for human tissue repair strategies. Previously we have shown that stem cells from olfactory mucosa are multipotent. As well, we have recently published three potential clinical applications. Their expression of dopaminergic markers in vitro and in a Parkinson’s rat transplant model has been demonstrated. Their conversion to chondrogenic phenotype in vitro and in vivo has also been described, as has their transplant into a rat model of cardiac infarction. Here we examine in detail the biology of the olfactory neural stem cell using the rat as our animal model cell source. We establish its presence by examining self-renewal capacity and for phenotypic acquisition in inductive circumstances. We determine its frequency within the cell population and show that our culture system selects for this putative stem cell. Our studies demonstrate that adult olfactory stem cells, when transplanted into an environmental niche different from that of their origin, are able to demonstrate multipotency by acquiring the phenotype of the resident cells. We investigate how immediate the instruction need be. We test the hypothesis that olfactory neurospheres contain stem cells whose capacity for differentiation is triggered by signals of the immediate environmental niche. Significantly, of importance to any tissue regeneration endeavor, stem cell numbers were shown to be enriched by our culture methods. This was confirmed whether measured by sphere-forming capacity or differentiation response rate.

Key words: Olfactory; Stem cell; Rat; Neurosphere; Clonal cell type; Cell differentiation; Multipotent

1Current address: King Faisal Specialist Hospital and Research Centre, P.O Box 3354 MBC 03, Riyadh 11211, Kingdom of Saudi Arabia.
Address correspondence to Wayne Murrell at his current address: Vilhelm Magnus Center, Institute for Surgical Research, Rikshospital, University of Oslo, Norway 0027. Tel: 47 23071405; Fax: 47 23071397; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1693–1705, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X565001
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparative Analysis of Telomere Length, Telomerase and Reverse Transcriptase Activity in Human Dental Stem Cells

Byeong-Gyun Jeon,* Eun-Ju Kang,* B. Mohana Kumar,* Geun-Ho Maeng,* Sun-A Ock,* Dae-Oh Kwack,† Bong-Wook Park,‡ and Gyu-Jin Rho*§

*OBS/Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Jinju, Republic of Korea
†Department of Biology Education, College of Education, Gyeongsang National University, Jinju, Republic of Korea
‡Department of Oral & Maxillofacial Surgery, School of Medicine and Institute of Health Sciences, Gyeongsang National University, Jinju, Republic of Korea
§Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea

Stem cells from dental tissues have been isolated and established for tooth regenerative applications. However, basic characterization on their biological properties still needs to be investigated before employing them for effective clinical trials. In this study, we compared the telomere length, relative telomerase activity (RTA), and relative reverse transcriptase activity (RRA) as well as the surface antigen profile and mesenchymal differentiation ability in human dental papilla stem cells (DPaSCs), dental pulp stem cells (DPuSCs), and dental follicle stem cells (DFSCs) with mesenchymal stem cells (MSCs) derived from bone marrow. Dental stem cells (DSCs) were strongly positive for cell surface markers, such as CD44 and CD90. However, slightly lower expression of CD105 was observed in DPaSCs and DPuSCs compared to DFSCs and MSCs. Following specific induction, DPaSCs, DFSCs, and MSCs were successfully differentiated into adipocytes and osteocytes. However, DPuSCS, in particular, were able to differentiate into adipocytes but failed to induce into osteogenic differentiation. Further, all DSCs, MSCs, and MRC-5 fibroblasts as control were investigated for telomere length by nonradioactive chemiluminescent assay, RTA by relative-quantitative telomerase repeat amplification protocol (RQ-TRAP), and RRA by PCR-based assay. Mean telomere lengths in DPaSCs, DPuSCs, DFSCs, and MSCs was ~11 kb, and the values did not differ significantly (p < 0.05) among the cells analyzed. RTA levels in DPaSCs were significantly (p < 0.05) higher than in MSCs, DPuSCs, DFSCs, and MRC-5 fibroblasts and among DSCs, DFSCs showed a significantly (p < 0.05) lower RTA. Moreover, RRA levels were significantly (p < 0.05) higher in DPaSCs, DPuSCs, and MSCs than in DFSCs. Based on these observations, we conclude that among DSCs, DPaSCs possessed ideal characteristics on telomere length, telomerase activity and reverse transcriptase (RTase) activity, and may serve as suitable alternative candidates for regenerative medicine.

Key words: Dental stem cells (DSCs); Mesenchymal stem cells (MSCs); Telomere length; Telomerase activity; Reverse transcriptase activity

Address correspondence to Gyu-Jin Rho, D.V.M., Ph.D., Professor, OBS/Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 900 Gazwa, Jinju, GN, Republic of Korea 660-701. Tel: +82-55-772-2347; Fax: +82-55-772-2349; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1707–1719, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566235
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Feeder-Free Derivation of Induced Pluripotent Stem Cells From Human Immature Dental Pulp Stem Cells

Patrícia C. B. Beltrão-Braga,*†‡ Graciela C. Pignatari,†‡ Paulo C. Maiorka,† Nélio A. J. Oliveira,§ Nelson F. Lizier,‡¶ Cristiane V. Wenceslau,†‡ Maria A. Miglino,†‡ Alysson R. Muotri,§ and Irina Kerkis‡¶

*School of Arts, Science and Humanities, Department of Obstetrics, University of São Paulo, São Paulo, Brazil
†Faculty of Veterinary Medicine, Department of Surgery and Department of Pathology, University of São Paulo, São Paulo, Brazil
‡National Institute of Science and Technology in Stem Cell and Cell Therapy (INCTC), Ribeirão Preto, Brazil
§Department of Pediatrics/Rady Children’s Hospital San Diego, Department of Cellular & Molecular Medicine, Stem Cell Program, School of Medicine, University of California San Diego, La Jolla, CA, USA
¶Butantan Institute, Laboratory of Genetics, São Paulo, Brazil

Induced pluripotent stem cells (iPSCs) can be created by forcing expression of certain genes in fibroblasts or other somatic cell types, reversing them to a pluripotent state similar to that of embryonic stem cells (ESC). Here, we used human immature dental pulp stem cells (hIDPSCs) as an alternative source for creating iPSC. hIDPSCs can be easily isolated from accessible tissue of young and adult patients. hIDPSCs possess a fibroblast-like morphology, retaining characteristics of adult multipotent stem cells. Reprogramming of hIDPSCs was fast, producing primary hIDPSC-iPSC colonies even under feeder-free conditions. hIDPSCs acquired ESC-like morphology, expressed pluripotent markers, possessed stable, normal karyotypes, and demonstrated the ability to differentiated in vitro and in vivo. Our data demonstrate that hIDPSCs-iPSCs offer an advantageous cell system for future cell therapy and basic studies, particularly as a model for pediatric developmental disorders.

Key words: Induced pluripotent stem cells (iPSCs); Pediatric diseases; Human immature dental pulp stem cells (hIDPSCs); Reprogramming

Address correspondence to Dr. Patricia C. B. Beltrão-Braga, School of Arts, Science and Humanities, Department of Obstetrics, University of São Paulo, 1000 Arlindo Bettio Av., 03828-000, São Paulo, Brazil. Tel: 55(11)3091-7690; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Dr. Alysson R. Muotri, Department of Pediatrics/Rady Children’s Hospital San Diego, Department of Cellular & Molecular Medicine, Stem Cell Program, School of Medicine, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, MC 0695, USA. Tel: 1-858-534-9320; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1721–1730, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X580590
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
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Surface Expression of HLA-G Is Involved in Mediating Immunomodulatory Effects of Placenta-Derived Multipotent Cells (PDMCs) Towards Natural Killer Lymphocytes

Ko-Jiunn Liu,*†‡ Chia-Jen Wang,§ Chan-Jung Chang,¶1 Hsin-I Hu,¶ Pei-Ju Hsu,¶ Yu-Chen Wu,* Chyi-Huey Bai,# Huey-Kang Sytwu,§**†† and B. Linju Yen¶‡‡§§

*National Institute of Cancer Research, National Health Research Institutes (NHRI), Tainan, Taiwan
†Institute of Biopharmaceutical Sciences, National Cheng Kung University, Tainan, Taiwan
‡School of Medical Laboratory Science and Biotechnology, Taipei Medical University (TMU), Taipei, Taiwan
§Department of Microbiology and Immunology, National Defense Medical Center (NDMC), Taipei, Taiwan
¶Regenerative Medicine Research Group, Institute of Cellular and System Medicine, National Health Research Institutes (NHRI), Zhunan, Taiwan
#Central Laboratory, Shin-Kong Wu Ho-Su Memorial Hospital and School of Public Health, Taipei Medical University (TMU), Taipei, Taiwan
**Graduate Institute of Life Sciences, National Defense Medical Center (NDMC), Taipei, Taiwan
††Department of Medical Research, Tri-Service General Hospital, National Defense Medical Center (NDMC), Taipei, Taiwan
‡‡Graduate Institute of Aging, College of Medicine, China Medical University, Taichung, Taiwan
§§Department of Obstetrics/Gynecology, Cathay General Hospital Shiji, Taipei, Taiwan

Interactions between maternal natural killer lymphocytes (NKs) and fetal tissues are important in mediating maternal–fetal tolerance. We therefore investigated the interactions of NKs to placenta-derived multipotent cells (PDMCs) isolated from the term human placenta. PDMCs have similar cell surface marker expression as bone marrow mesenchymal stem cells (BMMSCs) and additionally express human embryonic stem cell markers SSEA-4 and CD-9. Differentiation into the tri-mesodermal lineages of osteoblastic, adipocytic, and chondrogenic phenotypes can be readily achieved under the appropriate conditions. We found that PDMCs are more resistant to NK-mediated lysis than the major histocompatibility complex (MHC) class-I null target cell K562, and can suppress NK secretion of interferon-γ (IFN- γ). Moreover, as third-party cells, PDMCs suppressed the cytotoxic effects of cytokine-stimulated NKs on K562. Pretreatment of PDMCs with IFN- γ, a proinflammatory cytokine, surprisingly enhanced such immunosuppressive effects. Cell–cell contact between NKs and PDMCs is required for suppressive effects, which are partially mediated by slight upregulation of the NK inhibitory receptor killer inhibitory receptor and downregulation of the activating receptor NKp30. Moreover, enhancement of PDMC suppressive effects is also mediated by IFN- γ -induced surface expression of HLA-G—an immunomodulatory nonclassical MHC class I molecule—on PDMCs, as seen by partial reversibility with HLA-G neutralizing antibodies. With its broad immunosuppressive properties, PDMCs may represent a potential cell source for therapeutic use.

Key words: Placenta; Mesenchymal stem cells (MSCs); Natural killer lymphocytes; HLA-G; Immunomodulation

1Current address: Einstein Center for Human Embryonic Stem Cell Research, Albert Einstein College of Medicine, Bronx, NY, USA.
Address correspondence to B. Linju Yen, Regenerative Medicine Research Group, Institute of Cellular and System Medicine, National Health Research Institutes, No. 35, Keyan Road, Zhunan 35053, Taiwan. Tel: +886-37-246-166, ext. 37501; Fax: +886-37-587-408; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1731–1746, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X580536
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Establishment, Characterization, and Successful Adaptive Therapy Against Human Tumors of NKG Cell, a New Human NK Cell Line

Min Cheng,* Juan Ma,† Yongyan Chen,* Jianhua Zhang,‡ Weidong Zhao,† Jian Zhang,‡ Haiming Wei,* Bin Ling,† Rui Sun,* and Zhigang Tian*

*Institute of Immunology, Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, China
†Anhui Province Key Laboratory of Molecular Medicine, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China
‡Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan, China

Natural killer (NK) cells play important roles in adoptive cellular immunotherapy against certain human cancers. This study aims to establish a new human NK cell line and to study its role for adoptive cancer immunotherapy. Peripheral blood samples were collected from 54 patients to establish the NK cell line. A new human NK cell line, termed as NKG, was established from a Chinese male patient with rapidly progressive non-Hodgkin’s lymphoma. NKG cells showed LGL morphology and were phenotypically identified as CD56bright NK cell with CD16, CD27, CD3, αβTCR, γδTCR, CD4, CD8, CD19, CD161, CD45+, CXCR4+, CCR7+, CXCR1, and CX3CR1. NKG cells showed high expression of adhesive molecules (CD2, CD58, CD11a, CD54, CD11b, CD11c), an array of activating receptors (NKp30, NKp44, NKp46, NKG2D, NKG2C), and cytolysis-related receptors and molecules (TRAIL, FasL, granzyme B, perforin, IFN-γ). The cytotoxicity of NKG cells against tumor cells was higher than that of the established NK cell lines NK-92, NKL, and YT. NKG cell cytotoxicity depended on the presence of NKG2D and NKp30. When irradiated with 8 Gy, NKG cells were still with high cytotoxicity and activity in vitro and with safety in vivo, but without proliferation. Further, the irradiated NKG cells exhibited strong cytotoxicity against human primary ovarian cancer cells in vitro, and against human ovarian cancer in a mouse xenograft model. The adoptive transfer of NKG cells significantly inhibited the ovarian tumor growth, decreased the mortality rate and prolonged the survival, even in cases of advanced diseases. A number of NKG cells were detected in the ovarian tumor tissues during cell therapy. In use of the new human NK cell line, NKG would a promising cellular candidate for adoptive immunotherapy of human cancer.

Key words: Natural killer (NK) cell line; NKG cell; Adoptive transfer; Cancer immunotherapy; Human ovarian cancer

Address correspondence to Dr. Zhigang Tian, School of Life Sciences, University of Science and Technology of China, 443 Huangshan Road, Hefei, Anhui 230027, China. Tel: +86-551-360-7379; Fax: +86-551-360-6783; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1747–1758, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566217
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Time Is Crucial for Ex Vivo Expansion of T Regulatory Cells for Therapy

Natalia Marek,*† Maria Bieniaszewska,‡ Adam Krzystyniak,*† Jolanta Juścińska,§¶ Jolanta Myśliwska,# Piotr Witkowski,†** Andrzej Hellmann,‡ and Piotr Trzonkowski*†§

*Department of Clinical Immunology and Transplantology, Medical University of Gdańsk, Gdańsk Poland
†Department of Surgery, Section of Transplantation, University of Chicago, Chicago, IL, USA
‡Department of Hematology and Transplantology, Medical University of Gdańsk, Gdańsk Poland
§Tricity Academic Experimental Facility, Medical University of Gdańsk, Gdańsk Poland
¶Blood Bank of Pomerania Region, Gdańsk Unit, Gdańsk, Poland
#Department of Immunology, Medical University of Gdańsk, Gdańsk Poland
**Department of Surgery, Medical University of Gdańsk, Gdańsk Poland

Ex vivo expanded CD4+CD25highCD127 T regulatory cells (Tregs) are recognized as a promising candidate for immunosuppressive therapy in humans. However, due to the plasticity of Tregs lineage and artificial environment present during ex vivo expansion, Tregs easily lose suppressive activity. Here, we followed expanding CD4+CD25highCD127 Tregs and their naive (CD45RA+) and memory-like (CD45RA) subsets in order to establish the best conditions of the expansion. We found that, regardless of the phenotype sorted, expanding Tregs were undergoing changes resembling homeostatic proliferation and transformed into effector memory-like cells which produced not only suppressive interleukin-10 (IL-10) but also IL-6, IL-17, and interferon-γ (IFN-γ). With the time ex vivo, Tregs were losing the expression of FoxP3 and suppressive activity both when stimulated and when at rest. The only variable that helped preserve suppressive abilities of Tregs was the limitation of the time of ex vivo cultures to 2 weeks only. According to our study, the highest number of highly suppressive Tregs could be yielded with CD4+CD25highCD127 Tregs cultured no longer than 2 weeks. Thorough quality check, preferentially with the assessment of FoxP3 expression and IFN-γ suppression assay, should be applied to assess suppressive activity of the cells.

Key words: Cellular therapy; Immunotherapy; T regulatory cells (Tregs); Immunosuppression; Homeostatic proliferation

Address correspondence to Piotr Trzonkowski, M.D., Ph.D., Assistant Professor of Immunology, Department of Clinical Immunology and Transplantology, Medical University of Gdańsk, Ul. Dębinki 7, 80-211 Gdańsk, Poland. Tel: +48 58 3491590; Fax: +48 58 3491591; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1759–1769, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566244
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Rapamycin Generates Graft-Homing Murine Suppressor CD8+ T Cells That Confer Donor-Specific Graft Protection

Basset El Essawy,*† Prabhakar Putheti,* Wenda Gao,* and Terry B. Strom*

*Department of Medicine, Transplant Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA
†Department of Medicine, Al-Azhar University, Cairo, Egypt

It has been reported that rapamycin (RPM) can induce de novo conversion of the conventional CD4+Foxp3 T cells into CD4+Foxp3+ regulatory T cells (iTregs) in transplantation setting. It is not clear whether RPM can similarly generate suppressor CD8+ T cells to facilitate graft acceptance. In this study, we investigated the ability of short-term RPM treatment in promoting long-term acceptance (LTA) of MHC-mismatched skin allografts by generating a CD8+ suppressor T-cell population. We found that CD4 knockout (KO) mice (in C57BL/6 background, H-2b) can promptly reject DBA/2 (H-2d) skin allografts with mean survival time (MST) being 13 days (p < 0.01). However, a short course RPM treatment in these animals induced LTA with graft MST longer than 100 days. Adoptive transfer of CD8+ T cells from LTA group into recombination-activating gene 1 (Rag-1)-deficient mice provided donor-specific protection of DBA/2 skin grafts against cotransferred conventional CD8+ T cells. Functionally active immunoregulatory CD8+ T cells also resided in donor skin allografts. Eighteen percent of CD8+ suppressor T cells expressed CD28 as measured by flow cytometry, and produced reduced levels of IFN-γ, IL-2, and IL-10 in comparison to CD8+ effector T cells as measured by ELISA. It is unlikely that CD8+ suppressor T cells mediated graft protection via IL-10, as IL-10/Fc fusion protein impaired RPM-induced LTA in CD4 KO mice. Our data supported the notion that RPM-induced suppressor CD8+ T cells home to the allograft and exert donor-specific graft protection.

Key words: Rapamycin; CD8+ T cells; CD4 knockout; Transplantation; Rejection; Suppressor T cells

Address correspondence to Terry B. Strom, Department of Medicine, Transplant Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, E/CLS Room #608, Boston, MA 02215, USA. Tel: 1-617-735-2880; Fax: 1-617-735-2902; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1771–1790, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566172
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Functional Evaluation of Primary Renal Cell/Biomaterial Neo-Kidney Augment Prototypes for Renal Tissue Engineering

Joydeep Basu,* Christopher W. Genheimer,* Elias A. Rivera,* Richard Payne,* Kim Mihalko,† Kelly Guthrie,* Andrew T. Bruce,* Neil Robbins,* Darell McCoy,* Namrata Sangha,* Roger Ilagan,* Toyin Knight,* Thomas Spencer,* Belinda J. Wagner,* Manuel J. Jayo,* Deepak Jain,* John W. Ludlow,* and Craig Halberstadt*

*Tengion Inc., Winston-Salem, NC, USA
†Carolinas Medical Center, Charlotte, NC, USA

Development of a tissue-engineered neo-kidney augment (NKA) requires evaluation of defined, therapeutically relevant cell and cell/biomaterial composites (NKA constructs) for regenerative potential in mammalian kidney. Previous work identified primary renal cell populations that extended survival and improved renal function in a rodent model of chronic kidney disease (CKD). This study extends that work toward the goal of developing NKA by (i) screening in vivo inflammatory and fibrotic responses to acellular biomaterials delivered to healthy rodent renal parenchyma, (ii) evaluating the functionality of renal cell/biomaterial combinations in vitro, (iii) generating NKA constructs by combining therapeutically relevant cell populations with biocompatible biomaterial, and (iv) evaluating in vivo neokidney tissue development in response to NKA constructs delivered to healthy rodent renal parenchyma. Gelatin and hyaluronic acid (HA)-based hydrogels elicited the least inflammatory and fibrotic responses in renal parenchyma relative to polycaprolactone (PCL) and poly(lactic-co-glycolic acid) (PLGA) beads or particles and were associated with neovascularization and cellular infiltration by 4 weeks postimplantation. Renal cell populations seeded onto gelatin or HA-based hydrogels were viable and maintained a tubular epithelial functional phenotype during an in vitro maturation of 3 days as measured by transcriptomic, proteomic, secretomic, and confocal immunofluorescence assays. In vivo delivery of cell-seeded NKA constructs (bioactive renal cells + gelatin hydrogels) to healthy rodent renal parenchyma elicited neokidney tissue formation at 1 week postimplantation. To investigate a potential mechanism by which NKA constructs could impact a disease state, the effect of conditioned media on TGF-β signaling pathways related to tubulo-interstitial fibrosis associated with CKD progression was evaluated. Conditioned medium was observed to attenuate TGF-β-induced epithelial–mesenchymal transition (EMT) in vitro in a human proximal tubular cell line (HK2).

Key words: Kidney; Renal tissue engineering; Biomaterials; Regeneration; Neo-kidney augment

Address correspondence to Joydeep Basu, Tengion Inc., 3929 Westpoint Blvd., Winston-Salem, NC 27103, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1791–1803, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X564976
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
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Transplantation of Encapsulated Hepatocytes During Acute Liver Failure Improves Survival Without Stimulating Native Liver Regeneration

Antonino Sgroi,*† Gang Mai,*† Philippe Morel,* Reto M. Baertschiger,* Carmen Gonelle-Gispert,* Véronique Serre-Beinier,* and Leo H. Buhler*

*Surgical Research Unit, Department of Surgery, University Hospital Geneva, Geneva, Switzerland
†Department of General Surgery, West China Hospital, Sichuan University, Sichuan Province, China

The aim of this study was to evaluate the effects of intraperitoneal transplantation of encapsulated human hepatocytes on liver metabolism and regeneration of mice with acute liver failure. Primary human hepatocytes were immortalized using lentiviral vectors coding for antiapoptotic genes and microencapsulated using alginate-polylysine polymers. In vitro, immortalized human hepatocytes showed low, but stable, synthetic and catabolitic functions over time, when compared to primary hepatocytes. In vivo, mice with acute liver failure and transplanted with encapsulated immortalized human hepatocytes had a significantly improved survival and biochemical profile, compared to mice transplanted with empty capsules. Serum levels of cytokines implicated in liver regeneration were lower in mice transplanted with hepatocytes compared to mice receiving empty capsules. This decrease was significant for IL-6 and HGF at 3 h. Measurement of liver regeneration showed no significant difference between mice transplanted with hepatocytes compared to control groups. Intraperitoneal transplantation of encapsulated immortalized hepatocytes significantly improved survival of mice with acute liver failure by providing metabolic support and without modifying liver regeneration. The lower levels of cytokines implicated in liver regeneration suggest that the metabolic support provided by the encapsulated hepatocytes reduced the inflammatory stress on the liver and herein decreased the regenerative trigger on residual hepatocytes. These data emphasize that metabolic function and regeneration of hepatocytes are two distinct aspects that need to be studied and approached separately during acute liver failure.

Key words: Acute liver failure; Xenotransplantation; Encapsulation; Immortalized human hepatocytes; Liver regeneration; Hepatocyte transplantation

Address correspondence to Leo H. Buhler, M.D., Surgical Research Unit, Department of Surgery, University Hospital Geneva, 4, rue Gabrielle Perret-Gentil, 1211, Geneva 14, Switzerland. Tel: 41 22/3727698; Fax: 41 22/3727689; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1805–1815, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566154
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
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Endothelial Colony-Forming Cell Coating of Pig Islets Prevents Xenogeneic Instant Blood-Mediated Inflammatory Reaction

Jae Hyeon Kim,*† Bae Jun Oh,* Han Na Lee,† Ho Sun Park,† Sang Gyu Park,‡ and Kyong Soo Park†§

*Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea
†Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Republic of Korea
‡Department of Biomedical Science, CHA University, Seoul, Republic of Korea
§Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea

Instant blood-mediated inflammatory reaction (IBMIR) causes rapid islet loss in islet transplantation. Endothelial colony-forming cells (ECFCs) display unique abilities to promote angiogenesis and repair vascular injury compared to those of endothelial cells (ECs), which inhibits the allogeneic and xenogeneic IBMIR. We investigated the coating of pig islets with ex vivo-expanded ECFCs as a strategy to overcome xenogeneic IBMIR. Porcine islets were cocultured with human ECFCs in a specially modified culture medium for 2 days to obtain 70–90% coverage. The coating of pig islets with human ECFCs did not affect the glucosestimulated insulin secretion capacity or diabetes reversal rate after the transplantation of a marginal islet mass under the kidney capsules of diabetic nude mice compared to that of untreated islets. Uncoated islets, PBS control without islets, and the ECFC-coated islets were examined with an in vitro tubing loop assay using human blood. After 60 min of incubation in human blood, the ECFC-coated islets showed platelet consumption inhibition and low C3a and TAT assay results compared to those of the uncoated islets. Furthermore, there was very little macroscopic or microscopic clotting in the human ECFC-coated pig islets. The protective effect was more prominent compared to that of human EC coating of pig islets in our previous study. We investigated the changes in human-specific MCP-1, IL-8, and tissue factor (TF) levels after the coating of pig islets with human ECFCs or human ECs. The IL-8 levels after coating pig islets with ECFCs were significantly lower than those after coating pig islets with ECs, but there were no significant differences in the MCP-1 or TF levels between the ECFCs and ECs. In conclusion, the coating of pig islets with ECFCs completely prevented all components of xenogeneic IBMIR. ECFCs may be a better source of protection against xenogeneic IBMIR than are mature ECs.

Key words: Endothelial colony-forming cells (ECFCs); Islet; Pig; Endothelial cells (ECs)

Address correspondence to Kyong Soo Park, M.D., Ph.D., Department of Internal Medicine, Seoul National University College of Medicine, 28 Yongon-Dong Jongro-Gu, Seoul, 110-744, Republic Korea. Tel: +82-2-2072-1789; Fax: +82-2-3676-8309; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Jae Hyeon Kim, M.D., Ph.D., Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnamgu, 135-710, Seoul, Republic of Korea. Tel: +82-2-3410-1580; Fax: +82-2-3410-3849; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1817–1825, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X564994
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Highly Purified Versus Filtered Crude Collagenase: Comparable Human Islet Isolation Outcomes

Yong Wang,* Daniel Paushter,* Shusen Wang,* Barbara Barbaro,* Tricia Harvat,* Kirstie K. Danielson,† Katie Kinzer,* Liza Zhang,* Meirigeng Qi,* and Jose Oberholzer*

*Department of Transplant/Surgery, University of Illinois at Chicago, Chicago, IL, USA
†Division of Epidemiology & Biostatistics, University of Illinois at Chicago, Chicago, IL, USA

This study was designed to retrospectively compare the impact of crude Sigma V collagenase (Sigma V, n = 52) with high-purified Serva NB1 collagenase (Serva NB1, n = 42) on human islet isolation outcomes. A three-step filtration was applied to the crude Sigma V to remove endotoxin contamination and impurities; in addition, this process was used as a lot prescreening tool. Isolation outcomes were determined by digestion efficacy, islet yields, purity, viability, glucose-stimulated insulin release, and endotoxin content. The digestion efficacy between Sigma V and Serva NB1 was statistically significant (Sigma V: 64.71% vs. Serva NB1: 69.71%, p = 0.0014). However, the islet yields were similar (Sigma V: 23422.58 vs. Serva NB1: 271097 IEq, p = 0.23) between groups. There was no significant purity difference observed in fractions with purities greater than 75%. Viability (Sigma V: 93.3% vs. Serva NB1: 94.8%, p = 0.061) and stimulation indexes (Sigma V: 3.41 vs. Serva NB1: 2.74, p = 0.187) were also similar between the two groups. The impact of cold ischemia and age on the isolation outcome in the Sigma V group was comparable to the Serva NB1 group. The endotoxin content of the final products in the filtered Sigma V group was significantly less than that in the high-purified Serva NB1 group (0.022 vs. 0.052 EU/ml, p = 0.003). Additionally, in the Sigma V group there was minimal lot to lot variation and no significant loss of enzymatic activity after filtration. These findings indicate that the use of Sigma V or other crude enzyme blends for research pancreata is warranted to reduce isolation costs and increase the amount of islets available for critical islet research. These findings also validate the need for a systematic enzyme analysis to resolve these inconsistencies in overall enzyme quality once and for all.

Key words: Collagenase; Human islet isolation; Islet transplantation; Diabetes

Address correspondence to Jose Oberholzer, Department of Transplant/Surgery, University of Illinois at Chicago, Clinical Science Building, Suite 402, Chicago, IL 60612, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1827–1841, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X564085
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Characterization of Human Umbilical Cord Lining-Derived Epithelial Cells and Transplantation Potential

Yue Zhou,* Shu Uin Gan,† Gen Lin,‡ Yan Ting Lim,‡§ Jeyakumar Masilamani,†¶ Fatimah Bte Mustafa,‡ Meow Ling Phua,‡ Laura Rivino,‡ Toan Thang Phan,†¶ Kok Onn Lee,* Roy Calne,† and Paul A. MacAry‡

*Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
†Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
‡Immunology Program and Department of Microbiology, Yong Loo Lin School of Medicine, Centre for Life Sciences, National University of Singapore, Singapore
§Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore
¶CellResearch Corporation, Singapore

In this study we describe the derivation and immunological characterization of a primary epithelial cell type from the human umbilical cord membrane. These cord lining epithelial cells (CLECs) expressed and/or secreted isoforms of the nonclassical human leukocyte antigen class I (HLA-1b) glycoproteins, HLA-G and E. Conditioned media from CLECs inhibited mitogen-stimulated T-lymphocyte responses, and in a mixed leukocyte reaction (MLR) assay, cocultured CLECs inhibited allogeneic responses with a concomitant reduction in proinflammatory cytokines. Using a transwell coculture system, it was demonstrated that these immunoregulatory effects were mediated by soluble factors secreted by CLECs, in a dose-dependent manner. Functional studies using HLA-G blocking antibody showed that the effects of CLEC-secreted products could be inhibited, thus demonstrating a significant and important role for soluble HLA-G. In vivo, we show that transplanted CLECs could be maintained for extended periods in immunocompetent mice where xenorejection rapidly destroyed primary keratinocytes, a control human epithelial cell type. Additionally, CLECs delayed the rejection of keratinocytes and extended their survival when cotransplanted, indicating an ability to protect adjacent human cell types that would otherwise be rejected if transplanted alone. We also show that CLECs transduced with a modified human proinsulin gene were transplanted intraperitoneally into streptozotocin (STZ)-induced diabetic mice, resulting in significantly lower levels of serum glucose compared to control mice. This study has characterized the immunological properties of CLECs and tested a potential therapeutic application in the treatment of a type 1 diabetes mouse model.

Key words: Umbilical cord lining cell; HLA-G; Immune regulation; Tolerance; Cell transplantation

Address correspondence to Paul A. MacAry, LSI Immunology Program, Centre for Life Sciences, #03-05, 28 Medical Drive, Singapore 117456. Tel: 65-65165482; Fax: 65-67782684; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Roy Calne, Department of Surgery, National University of Singapore c/o National University Hospital, Service Block, Basement 1, 5 Lower Kent Ridge Road, Singapore 119074. Tel/fax: 65-68741913; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1843–1854, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X565038
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Intratracheal Transplantation of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells Dose-Dependently Attenuates Hyperoxia-Induced Lung Injury in Neonatal Rats

Yun Sil Chang,*†1 Soo Jin Choi,‡1 Dong Kyung Sung,† Soo Yoon Kim,† Wonil Oh,‡ Yoon Sun Yang,‡ and Won Soon Park*†

*Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea
†Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Seoul, Korea
‡Biomedical Research Institute, MEDIPOST Co., Ltd., Seoul, Korea

Intratracheal transplantation of human umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) attenuates the hyperoxia-induced neonatal lung injury. The aim of this preclinical translation study was to optimize the dose of human UCB-derived MSCs in attenuating hyperoxia-induced lung injury in newborn rats. Newborn Sprague-Dawley rats were randomly exposed to hyperoxia (95% oxygen) or normoxia after birth for 14 days. Three different doses of human UCB-derived MSCs, 5 × 103 (HT1), 5 × 104 (HT2), and 5 × 105 (HT3), were delivered intratracheally at postnatal day (P) 5. At P14, lungs were harvested for analyses including morphometry for alveolarization, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining, myeoloperoxidase activity, mRNA level of tumor necross factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and transforming growth factor-β  (TGF-β), human glyceradehyde-3-phosphate dehydrogenase (GAPDH), and p47phox, and collagen levels. Increases in TUNEL-positive cells were attenuated in all transplantation groups. However, hyperoxia-induced lung injuries, such as reduced alveolarization, as evidenced by increased mean linear intercept and mean alveolar volume, and increased collagen levels were significantly attenuated in both HT2 and HT3, but not in HT1, with better attenuation in HT3 than in HT2. Dose-dependent human GAPDH expression, indicative of the presence of human RNA in lung tissue, was observed only in the transplantation groups, with higher expression in HT3 than in HT2, and higher expression in HT2 than in HT1. Hyperoxia-induced inflammatory responses such as increased myeloperoxidase acitivity, mRNA levels of TNF-α, IL-1β, IL-6, and TGF-β  of the lung tissue, and upregulation of both cytosolic and membrane p47 phox, indicative of oxidative stress, were significantly attenuated in both HT2 and HT3 but not in HT1. These results demonstrate that intratracheal transplantation of human UCB-derived MSCs with appropriate doses may attenuate hyperoxia-induced lung injury through active involvement of these cells in modulating host inflammatory responses and oxidative stress in neonatal rats.

Key words: Bronchopulmonary dysplasia; Cell transplantation; Inflammation; Animal; Newborn

1Co-first authors.
Address correspondence to Won Soon Park, M.D., Department of Pediatrics, Samsung Medical Center; Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul 135-710, Korea. Tel: +82.2-3410-3523; Fax: +82.2-3410-0043; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1855–1866, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368910X557236
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Neural Differentiation of Brain-Derived Neurotrophic Factor-Expressing Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Culture via TrkB-Mediated ERK and β-Catenin Phosphorylation and Following Transplantation Into the Developing Brain

Jung Yeon Lim,* Sang In Park,* Seong Muk Kim,* Jin Ae Jun,* Ji Hyeon Oh,* Chung Hun Ryu,* Chang Hyun Jeong,* Sun Hwa Park,* Soon A. Park,* Wonil Oh,† Jong Wook Chang,† and Sin-Soo Jeun*‡

*Department of Biomedical Science, College of Medicine, The Catholic University of Korea, Seoul, Korea
†Medipost Biomedical Research Institute, MEDIPOST Co., Ltd., Seoul, Korea
‡Department of Neurosurgery, Seoul St. Mary’s Hospital, The Catholic University of Korea, Seoul, Korea

The ability of mesenchymal stem cells (MSCs) to differentiate into neural cells makes them potential replacement therapeutic candidates in neurological diseases. Presently, overexpression of brain-derived neurotrophic factor (BDNF), which is crucial in the regulation of neural progenitor cell differentiation and maturation during development, was sufficient to convert the mesodermal cell fate of human umbilical cord blood-derived MSCs (hUCB-MSCs) into a neuronal fate in culture, in the absence of specialized induction chemicals. BDNF overexpressing hUCB-MSCs (MSCs-BDNF) yielded an increased number of neuron-like cells and, surprisingly, increased the expression of neuronal phenotype markers in a time-dependent manner compared with control hUCB-MSCs. In addition, MSCs-BDNF exhibited a decreased labeling for MSCs-related antigens such as CD44, CD73, and CD90, and decreased potential to differentiate into mesodermal lineages. Phosphorylation of the receptor tyrosine kinase B (TrkB), which is a receptor of BDNF, was increased significantly in MSC-BDNF. BDNF overexpression also increased the phosphorylation of β-catenin and extracellular signal-regulated kinases (ERKs). Inhibition of TrkB availability by treatment with the TrkBspecific inhibitor K252a blocked the BDNF-stimulated phosphorylation of β-catenin and ERKs, indicating the involvement of both the β-catenin and ERKs signals in the BDNF-stimulated and TrkB-mediated neural differentiation of hUCB-MSCs. Reduction of β-catenin availability using small interfering RNA-mediated gene silencing inhibited ERKs phosphorylation. However, β-catenin activation was maintained. In addition, inhibition of β-catenin and ERKs expression levels abrogated the BDNF-stimulated upregulation of neuronal phenotype markers. Furthermore, MSC-BDNF survived and migrated more extensively when grafted into the lateral ventricles of neonatal mouse brain, and differentiated significantly into neurons in the olfactory bulb and periventricular astrocytes. These results indicate that BDNF induces the neural differentiation of hUCB-MSCs in culture via the TrkB-mediated phosphorylation of ERKs and β-catenin and following transplantation into the developing brain.

Key words: Brain-derived neurotrophic factor (BDNF); β-Catenin; Extracellular signal-regulated kinases (ERKs); Mesenchymal stem cells (MSCs); Neural differentiation; Tyrosine kinase B (TrkB)

Address correspondence to Sin-Soo Jeun, Department of Biomedical Science, College of Medicine, The Catholic University of Korea, Seoul, Korea. Tel: 82-2-2258-7535; Fax: 82-2-3482-1853; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1867–1880, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566163
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of Canine Umbilical Cord Blood-Derived Mesenchymal Stem Cell Transplantation Times: Involvement of Astrogliosis, Inflammation, Intracellular Actin Cytoskeleton Pathways, and Neurotrophin-3

Sung-Su Park,* Ye-Eun Byeon,* Hak-Hyun Ryu,* Byung-Jae Kang,* YongSun Kim,* Wan-Hee Kim,* Kyung-Sun Kang,† Ho-Jae Han,‡ and Oh-Kyeong Kweon*

*Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University, Seoul, Korea
†Laboratory of Stem cell and Tumor Biology, Department of Veterinary Public Health, College of Veterinary Medicine, Seoul National University, Seoul, Korea
‡Department of Veterinary Physiology, College of Veterinary Medicine, Seoul National University, Seoul, Korea

Canine mesenchymal stem cells (cMSCs) derived from umbilical cord blood represent a potentially useful source of stem cells for therapy. The aim of this study was to compare the effects of different transplantation times of cMSCs after spinal cord injury (SCI). A total of 21 dogs were subjected to SCI by balloon-induced compression of the first lumbar vertebrae for 12 h. Of the 21 dogs, 12 were divided into four groups of three according to the time of stem cell (1 × 106) transplantation at the injury site: control no treatment, 12 h, 1 week, and 2 weeks. The remaining 9 animals were negative harvest (HA) time controls for each treatment group (n = 3). Olby and Tarlov scores were used to evaluate functional recovery of the hindlimbs. Markers for neuronal regeneration (Tuj-1, nestin, MAP2, and NF-M), astrogliosis (GALC, GFAP, and pSTAT3), signal molecules for actin cytoskeleton (RhoA, Cdc42, and Rac1), inflammation (COX-2), and neurotrophins (NT-3) were evaluated by Western blot analysis. Scores of the 1-week transplantation group showed significant improvement compared to controls. Hematoxylin and eosin (H&E) staining revealed less fibrosis at the injury site in the 1-week transplantation group compared to other groups and immunohistochemistry showed increased expression of neuronal markers. Furthermore, in both 1-week and 2-week transplantation groups, Tuj-1, nestin, MAP2, NF-M, NT-3, and GFAP increased, but pSTAT3, GALC, and COX2 decreased. RhoA decreased and Rac1 and Cdc42 increased in the 1-week transplantation group. In conclusion, transplantation of cMSCs 1 week after SCI was more effective in improving clinical signs and neuronal regeneration and reducing fibrosis formation compared to the other transplantation times evaluated. Subsequently, these data may contribute to the optimization of timing for MSC transplantation used as a therapeutic modality.

Key words: Dog; Stem cells; Optimal transplantation time; Spinal cord injury

Address correspondence to Oh-Kyeong Kweon, Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. Tel: +82-2-880-1248; Fax: +82-2-888-2866; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1881–1899, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566181
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bone Marrow Mesenchymal Stem Cells in a Three-Dimensional Gelatin Sponge Scaffold Attenuate Inflammation, Promote Angiogenesis, and Reduce Cavity Formation in Experimental Spinal Cord Injury

Xiang Zeng,* Yuan-shan Zeng,*† Yuan-huan Ma,* Li-ya Lu,† Bao-ling Du,† Wei Zhang,† Yan Li,† and Wood Yee Chan‡

*Research center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou, China
†Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
‡School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China

Three-dimensional (3D) gelatin sponge (GS) scaffolds were constructed by ensheathing GS with a thin film of poly-(lactide-co-glycolide) (PLGA). Rat bone marrow-derived mesenchymal stem cells (MSCs) were isolated, cultured, and then seeded to the scaffolds. Distribution of cells and cell growth, survival, and proliferation within the scaffolds were then determined. Immunofluorescence and Western blot analysis were employed to detect the deposition of fibronectin to the scaffolds on day 3 and day 7 of culture. Scaffolds with or without MSCs were then transplanted into the transected rat spinal cord. One or 8 weeks following transplantation, cavity areas, activated macrophages/microglia, expression of TNF-α and IL-1β, and neovascularization within the grafts were examined and quantified. Deposition of fibronectin (FN) and expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α  (HIF-1α) as potential inducing factors for angiogenesis were also examined. Results showed that 3D GS scaffolds allowed MSCs to adhere, survive, and proliferate and also FN to deposit. In vivo transplantation experiments demonstrated that these scaffolds were biocompatible, and MSCs seeded to the scaffolds played an important role in attenuating inflammation, promoting angiogenesis, and reducing cavity formation. Therefore, the GS scaffolds with MSCs may serve as promising supporting transplants for repairing spinal cord injury.

Key words: Gelatin sponge; Mesenchymal stem cells (MSCs); Inflammation; Angiogenesis; Tissue engineering; Spinal cord injury

Address correspondence to Yuan-Shan Zeng, M.D., Ph.D., Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yatsen University, 74# Zhongshan 2nd Road, Guangzhou 510080, China. Tel: 001-86-20-87332698; Fax: 001-86-20-87332698; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Wood Yee Chan, Ph.D., School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong, China. Tel: 852-3943 6895; Fax: 852-2603 5031; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1901–1906, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566190
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Injection Parameters Affect Cell Viability and Implant Volumes in Automated Cell Delivery for the Brain

Douglas Kondziolka,*† Glenn T. Gobbel,*† Wendy Fellows-Mayle,* Yue-Fang Chang,* and Martin Uram‡

*Department of Neurological Surgery, University of Pittsburgh, Pittsburgh, PA, USA
†McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA
‡Corporate Innovations Department, MEDRAD, Inc., Indianola, PA, USA

The technique of central nervous system cell implantation can affect the outcome of preclinical or clinical studies. Our goal was to evaluate the impact of various injection parameters that may be of consequence during the delivery of solute-suspended cells. These parameters included (1) the type and concentration of cells used for implantation, (2) the rate at which cells are injected (flow rate), (3) the acceleration of the delivery device, (4) the period of time between cell loading and injection into the CNS (delay), and (5) the length and gauge of the needle used to deliver the cells. Neural progenitor cells (NPCs) and bone marrow stromal cells (BMSCs) were injected an automated device. These parameters were assessed in relation to their effect on the volume of cells injected and cell viability. Longer and thinner cannulae and higher cell concentrations were detrimental for cell delivery. Devices and techniques that optimize these parameters should be of benefit.

Key words: Brain; Cell transplantation; Stroke; Stem cell

Address correspondence to Douglas Kondziolka, M.D., M.Sc., FRCSC, Department of Neurological Surgery, Suite B-400, UPMC Presbyterian, 200 Lothrop Street, Pittsburgh, PA 15213, USA. Tel: 412-647-6782; Fax: 412-6476483; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1907–1914, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X565038
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Implantation of Sinoatrial Node Cells Into Canine Right Ventricle: Biological Pacing Appears Limited by the Substrate

Hao Zhang,* David H. Lau,† Iryna N. Shlapakova,† Xin Zhao,‡ Peter Danilo,† Richard B. Robinson,† Ira S. Cohen,§ Dan Qu,* Zhiyun Xu,* and Michael R. Rosen†¶

*Changhai Hospital, Second Military Medical University, Shanghai, China
†Department of Pharmacology, Center for Molecular Therapeutics, College of Physicians and Surgeons, Columbia University, New York, NY, USA
‡Division of Cardiology, First Affiliated Hospital of Soochow University, Su Zhou, China
§Department of Physiology & Biophysics, Stony Brook University, Stony Brook, NY, USA
¶Department of Pediatrics, College of Physicians and Surgeons, Columbia University, New York, NY, USA

Biological pacing has been proposed as a physiologic counterpart to electronic pacing, and the sinoatrial node (SAN) is the general standard for biological pacemakers. We tested the expression of SAN pacemaker cell activity when implanted autologously in the right ventricle (RV). We induced complete heart block and implanted electronic pacemakers in the RV of adult mongrel dogs. Autologous SAN cells isolated enzymatically were studied by patch clamp to confirm SAN identity. SAN cells (400,000) were injected into the RV subepicardial free wall and dogs were monitored for 2 weeks. Pacemaker function was assessed by overdrive pacing and IV epinephrine challenge. SAN cells expressed a time-dependent inward current (If) activating on hyperpolarization: density = 4.3 ± 0.6 pA/pF at −105 mV. Four of the six dogs demonstrated >50% of beats originating from the implant site at 24 h. Biological pacemaker rates on days 7–14 = 45–55 bpm and post-overdrive escape times = 1.5–2.5 s. Brisk catecholamine responsiveness occurred. Dogs implanted with autologous SAN cells manifest biological pacing properties dissimilar from those of the anatomic SAN. This highlights the importance of cell and substrate interaction in generating biological pacemaker function.

Key words: Pacemaker current; Heart block; Cardiac pacing; Sinoatrial node

Address correspondence to Michael R. Rosen, M.D., Gustavus A. Pfeiffer Professor of Pharmacology, Professor of Pediatrics, Columbia University, 630 West 168 Street, PH7West-321, New York, NY, 10032, USA. Tel: 212-305-8754; Fax: 212-305-8351; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Zhiyun Xu, M.D., Director, Professor, Institute of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, 168 Changhai Road, Shanghai, 200433, China. Tel: 13818178916; Fax: 86-021-65490979; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 20, pp. 1915–1920, 2011
0963-6897/11 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X566253
E-ISSN 1555-3892
Copyright © 2011 Cognizant Comm. Corp.
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Brief Communication

Human CD47 Expression Permits Survival of Porcine Cells in Immunodeficient Mice That Express SIRPα Capable of Binding to Human CD47

Chunfeng Wang,*†1 Hui Wang,†‡1 Kentaro Ide,§ Yuantao Wang,† Nico Van Rooijen,¶ Hideki Ohdan,§ and Yong-Guang Yang†‡

*College of Animal Science and Technology, Jilin Agricultural University, Changchun, China
†Transplantation Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
‡Columbia Center for Translational Immunology, Columbia University College of Physicians and Surgeons, New York, NY, USA
§Department of Surgery, Division of Frontier Medical Science, Programs for Biomedical Research, Graduate School of Biomedical Science, Hiroshima University, Hiroshima, Japan
¶Department of Cell Biology, Vrije Universiteit, Amsterdam, The Netherlands

Signal regulatory protein α (SIRPα) is a critical immune inhibitory receptor on macrophages, and its interaction with CD47 prevents autologous phagocytosis. We have previously shown that pig CD47 does not interact with human SIRPα, and that human CD47 expression inhibits phagocytosis of porcine cells by human macrophages in vitro. In this study, we have investigated the potential of human CD47 expression to promote porcine cell survival in vivo. Human CD47-expressing and control porcine B-lymphoma cells were transplanted into T- and B-cell-deficient nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice that express SIRPα  capable of interacting with human CD47. Only the human CD47-expressing porcine lymphoma cells survived and were able to form tumors in NOD/SCID mice; however, both the control and human CD47-expressing porcine cells survived in macrophage-depleted NOD/SCID mice. These results indicate that transgenic expression of human CD47 may provide an effective approach to inhibiting macrophage-mediated xenograft rejection in clinical xenotransplantation.

Key words: CD47; Macrophage; Pig; Signal regulatory protein α (SIRPα); Xenotransplantation

1These authors provided equal contribution to this work.
Address correspondence to Yong-Guang Yang, M.D., Ph.D., Columbia Center for Translational Immunology, Columbia University College of Physicians and Surgeons, 630 West 168th St, PS17-514, New York, NY 10032, USA. Tel: (212) 304-5586; Fax: (646) 426-0036; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it