Call Transplantation 23(8) Abstracts

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Cell Transplantation, Vol. 23, pp. 921-928, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X666412
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Effects of Donor-, Pancreas-, and Isolation-Related Variables on Human Islet Isolation Outcome: A Systematic Review

Denise E. Hilling,* Eelco Bouwman,* Onno T. Terpstra,* and Perla J. Marang-van de Mheen†

*Department of Surgery, Leiden University Medical Center, Leiden, the Netherlands
†Department of Medical Decision Making, Leiden University Medical Center, Leiden, the Netherlands

Different factors have been reported to influence islet isolation outcome, but their importance varies between studies and are hampered by the small sample sizes in most studies. The purpose of this study was to perform a systematic review to assess the impact of donor-, pancreas-, and isolation-related variables on successful human islet isolation outcome. PubMed, Embase, and Web of Science were searched electronically in April 2009. All studies reporting on donor-, pancreas-, and isolation-related factors relating to prepurification and postpurification islet isolation yield and proportion of successful islet isolations were selected. Seventy-four retrospective studies had sufficient data and were included in the analyses. Higher pre- and postpurification islet yields and a higher proportion of successful islet isolations were obtained when pancreata were preserved with the twolayer method rather than University of Wisconsin solution in donors with shorter cold ischemia times (CITs) [1 h longer CIT resulted in an average decline of prepurification and postpurification yields and proportion of successful isolations of 59 islet equivalents (IEQs)/g, 54 IEQs/g, and 21%, respectively]. Higher prepurification yields and higher percentage of successful islet isolations were found in younger donors with higher body mass index. Lower yields were found in donation after brain death donors compared to donation after cardiac death donors. Higher postpurification yields were found for isolation with Serva collagenase. This review identified donor-, pancreas-, and isolation-related factors that influence islet isolation yield. Standardized reports of these factors in all future studies may improve the power and identify additional factors and thereby contribute to improving islet isolation yield.

Key words: Islet isolation; Outcome; Human; Donor; Pancreas

Received February 24, 2012; final acceptance May 23, 2013. Online prepub date: April 29, 2013.
Address correspondence to Denise E. Hilling, M.D., Ph.D., Department of Surgery, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, the Netherlands. Tel: +31-71-5264574; Fax: +31-71-5266838; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 929-944, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667033
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Enhancement of In Vitro and In Vivo Function of Agarose-Encapsulated Porcine Islets by Changes in the Islet Microenvironment

Robert W. Holdcraft,* Lawrence S. Gazda,*† Lisa Circle,* Hollie Adkins,* Steven G. Harbeck,* Eric D. Meyer,* Melissa A. Bautista,* Prithy C. Martis,* Melissa A. Laramore,* Horatiu V. Vinerean,* Richard D. Hall,‡ and Barry H. Smith†§¶

*The Rogosin Institute–Xenia Division, Xenia, OH, USA
†The Rogosin Institute, New York, NY, USA
‡Bob Evans Farms, Inc., Columbus, OH, USA
§Department of Surgery, Weill Medical College of Cornell University, New York, NY, USA
¶NewYork-Presbyterian Hospital, New York, NY, USA

The transplantation of porcine islets of Langerhans to treat type 1 diabetes may provide a solution to the demand for insulin-producing cells. Porcine islets encapsulated in agarose–agarose macrobeads have been shown to function in nonimmunosuppressed xenogeneic models of both streptozotocin-induced and autoimmune type 1 diabetes. One advantage of agarose encapsulation is the ability to culture macrobeads for extended periods, permitting microbiological and functional assessment. Herein we describe optimization of the agarose matrix that results in improved islet function. Porcine islets (500 IEQs) from retired breeding sows were encapsulated in 1.5% SeaKem Gold (SG), 0.8% SG, or 0.8% Litex (Li) agarose, followed by an outer capsule of 5% SG agarose. Insulin production by the encapsulated islets exhibited an agarose-specific effect with 20% (0.8% SG) to 50% (0.8% Li) higher initial insulin production relative to 1.5% SG macrobeads. Insulin production was further increased by 40–50% from week 2 to week 12 in both agarose types at the 0.8% concentration, whereas islets encapsulated in 1.5% SG agarose increased insulin production by approximately 20%. Correspondingly, fewer macrobeads were required to restore normoglycemia in streptozotocin-induced diabetic female CD(SD) rats that received 0.8% Li (15 macrobeads) or 0.8% SG (17 macrobeads) as compared to 1.5% SG (19 macrobeads). Islet cell proliferation was also observed during the first 2 months postencapsulation, peaking at 4 weeks, where approximately 50% of islets contained proliferative cells, including b-cells, regardless of agarose type. These results illustrate the importance of optimizing the microenvironment of encapsulated islets to improve islet performance and advance the potential of islet xenotransplantation for the treatment of type 1 diabetes.

Key words: Porcine islets; Macrobeads; Encapsulation; Xenotransplantation

Received March 30, 2012; final acceptance April 15, 2013. Online prepub date: April 29, 2013.
Address correspondence to Robert W. Holdcraft, 740 Birch Road, Xenia, OH 45385, USA. Tel: +1-937-374-3116; Fax: +1-937-374-3261; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 945-957, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X670183
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Neonatal Human Dermal Fibroblasts Immobilized in RGD–Alginate Induce Angiogenesis

Susana G. Guerreiro,*†‡ Maria J. Oliveira,*§ Mario A. Barbosa,*‡¶ Raquel Soares,† and Pedro L. Granja*‡

*INEB – Instituto de Engenharia Biomedica, Universidade do Porto, Porto, Portugal
†Universidade do Porto, Faculdade de Medicina (FMUP), Departamento de Bioquimica, Porto, Portugal
‡Universidade do Porto, Faculdade de Engenharia (FEUP), Porto, Portugal
§Universidade do Porto, Faculdade de Medicina (FMUP), Departamento de Anatomia Patologica, Porto, Portugal
¶Universidade do Porto, Instituto de Ciencias Biomedicas Abel Salazar (ICBAS), Porto, Portugal

Promoting angiogenesis in a damaged tissue is a major challenge for tissue regeneration. Recent findings in tissue engineering suggest that fibroblasts (FBs) play an important role in orchestrating the angiogenic process. Fibroblasts maintain the structural integrity of connective tissue by continuously secreting growth factors and extracellular matrix precursors, which are essential for endothelial cell (EC) adhesion and spreading, thus playing a crucial role in angiogenesis. We hypothesized that FBs immobilized in alginate gels grafted with the RGD peptidic sequence could influence the recruitment of ECs to improve vascularization. In this work, the modulation of immobilized human FBs within the 3D synthetic extracellular matrix was assessed. Experiments using cocultures of ECs and FBs in indirect contact as well as angiogenic assays were performed to assess the influence of FBs immobilized in RGD–alginate in ECs’ viability, stabilization, sprouting, and assembly into capillary-like structures. This study demonstrates the ability of FBs immobilized within RGD–alginate microspheres to modulate and support capillary-like structures’ assembly. These findings indicate that the microenvironment created by these stromal cells in the scaffold modulates capillary morphogenesis, thus stimulating angiogenesis in situ and can potentially be used in regenerative medicine in clinical scenarios where vascularization is essential.

Key words: Alginate; Angiogenesis; Endothelial cells (ECs); Fibroblasts (FBs); RGD peptide: Injectable biomaterials

Received November 11, 2011; final acceptance July 20, 2012. Online prepub date: July 17, 2013.
Address correspondence to Pedro L. Granja, INEB – Instituto de Engenharia Biomedica, Universidade do Porto, Rua do Campo Alegre, 823, 4150-180 Porto, Portugal. Tel: +351-226074900; Fax: +351-226094567; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 959-979, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667006
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Umbilical Cord Blood-Derived CD34+ Cells Improve Outcomes of Traumatic Brain Injury in Rats by Stimulating Angiogenesis and Neurogenesis

Sheng-Hsien Chen,*† Jhi-Joung Wang,‡ Chung-Hwan Chen,§ Hsiu-Kang Chang,¶ Mao-Tsun Lin,‡ Fong-Ming Chang,# and Chung-Ching Chio**

*Da-An Hospital of Women and Children, Tainan, Taiwan
†Department of Biotechnology, Southern Taiwan University of Science and Technology, Tainan, Taiwan
‡Department of Medical Research, Chi Mei Medical Center, Tainan, Taiwan
§Department of Orthopedics, Kaohsiung Medical University, Kaohsiung, Taiwan
¶Stem Cell Research Center, Health Banks Biotech Co., Ltd., Taipei, Taiwan
#Department of Obstetrics and Gynecology, National Cheng-Kung University School of Medicine, Tainan, Taiwan
**Department of Surgery, Chi Mei Medical Center, Tainan, Taiwan

Human umbilical cord blood cells (HUCBCs) have been shown to be beneficial in reducing neurological deficits in rats with brain fluid percussion injury (FPI). This study aimed to assess the basic mechanisms underlying the neuroprotective effects of HUCBC-derived cluster of differentiation 34-positive (CD34+) cells. Rats were divided into three major groups: (i) sham-operated controls; (ii) FPI rats treated with phosphate-buffered saline (PBS); (iii) FPI rats treated with 0.2%, 50%, or 95% CD34+ cells (in 5 × 105 cord blood lymphocytes and monocytes). Intravenous (IV) administration of 0.3 ml of PBS, 0.2% CD34+ cells, 50% CD34+ cells, or 95% CD34+ cells was conducted immediately after FPI. It was found that 4 days post-FPI, CD34+ cells could be detected in the ischemic brain tissues for 50% CD34+ cell- or 95% CD34+ cell-treated FPI rats, but not for the PBS-treated FPI rats or the 0.2% CD34+ cell-treated FPI rats. CD34+ cell (0.2%)-treated FPI rats or PBS-treated FPI rats displayed neurological and motor deficits, cerebral contusion and apoptosis [e.g., increased numbers of both TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling)-positive cells and caspase-3-positive cells], and activated inflammation (e.g., increased serum levels of tumor necrosis factor-a). FPI-induced neurological motor dysfunction, cerebral contusion and apoptosis, and activated inflammation could be attenuated by 50% CD34+ or 95% CD34+ cell therapy. In addition 50% or 95% CD34+ cell therapy but not PBS or 0.2% CD34+ cell therapy significantly promoted angiogenesis (e.g., increased numbers of both vasculoendothelial growth factor-positive cells and 5-bromodeoxyuridine (BrdU)-endothelial double-positive cells), neurogenesis (e.g., increased numbers of both glial cell line-derived neurotrophic factor-positive cells and BrdU/neuronal nuclei double-positive cells) in the ischemic brain after FPI, and migration of endothelial progenitor cells from the bone marrow. Our data suggest that IV administration of HUCBC-derived CD34+ cells may improve outcomes of FPI in rats by stimulating both angiogenesis and neurogenesis.

Key words: Traumatic brain injury; CD34+ cells; Apoptosis; Cytokines; Angiogenesis; Neurogenesis; Progenitor cells

Received July 3, 2012; final acceptance April 9, 2013. Online prepub date: April 12, 2013.
Address correspondence to Dr. Chung-Ching Chio, M.D., Department of Surgery or Dr. Mao-Tsun Lin, Ph.D., Department of Medical Research, Chi Mei Medical Center, Tainan 710, Taiwan. Tel: +886 6 2812811, ext: 52657; Fax: +886 6 2832639; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 981-994, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X664865
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Survival and Integration of Neurons Derived From Human Embryonic Stem Cells in MPTP-Lesioned Primates

Dustin R. Wakeman,* Stephanie Weiss,† John R. Sladek, Jr.,‡§ John D. Elsworth,† Brian Bauereis,† Csaba Leranth,¶# Patrick J. Hurley,* Robert H. Roth,* and D. Eugene Redmond, Jr.†**††

*Department of Neurological Sciences, Rush University Medical Center, Chicago, IL, USA
†Department of Psychiatry, Yale University School of Medicine, New Haven, CT, USA
‡Department of Neurology, University of Colorado Health Sciences Center, Denver, CO, USA
§Department of Pediatrics, University of Colorado Health Sciences Center, Denver, CO, USA
¶Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, CT, USA
#Department of Neurobiology, Yale University School of Medicine, New Haven, CT, USA
**Department of Neurosurgery, Yale University School of Medicine, New Haven, CT, USA
††St. Kitts Biomedical Research Foundation, St. Kitts-Nevis, West Indies

A human embryonic stem cell (HESC) line, H1, was studied after differentiation to a dopaminergic phenotype in vitro in order to carry out in vivo studies in Parkinsonian monkeys. To identify morphological characteristics of transplanted donor cells, HESCs were transfected with a GFP lentiviral vector. Gene expression studies were performed at each step of a neural rosette-based dopaminergic differentiation protocol by RT-PCR. In vitro immunofluorescence revealed that >90% of the differentiated cells exhibited a neuronal phenotype by β-III-tubulin immunocytochemistry, with 17% of the cells coexpressing tyrosine hydroxylase prior to implantation. Biochemical analyses demonstrated dopamine release in culture in response to potassium chloride-induced membrane depolarization, suggesting that the cells synthesized and released dopamine. These characterized, HESC-derived neurons were then implanted into the striatum and midbrain of MPTP (1-methyl-4- phenyl-1,2,3,6-tetrahydropyridine)-exposed monkeys that were triple immunosuppressed. Here we demonstrate robust survival of transplanted HESC-derived neurons after 6 weeks, as well as morphological features consistent with polarization, organization, and extension of processes that integrated into the host striatum. Expression of the dopaminergic marker tyrosine hydroxylase was not maintained in HESC-derived neural grafts in either the striatum or substantia nigra, despite a neuronal morphology and expression of β-III-tubulin. These results suggest that dopamine neuronal cells derived from neuroectoderm in vitro will not maintain the correct midbrain phenotype in vivo in nonhuman primates, contrasted with recent studies showing dopamine neuronal survival using an alternative floorplate method.

Key words: Embryonic stem cells; Neural differentiation; Stem cell transplantation; Primates; MPTP; Parkinson’s disease; Dopamine

Received October 28, 2012; final acceptance February 26, 2013. Online prepub date: April 2, 2013.
Address correspondence to D. E. Redmond, Jr., 300 George St., Suite 9-32, New Haven, CT., 06510 USA. Tel: 203-785-4432; Fax: 203-785-5416; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 995-1007, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X666449
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Transplantation of Human Fetal Tissue for Neurodegenerative Diseases: Validation of a New Protocol for Microbiological Analysis and Bacterial Decontamination

Tobias Piroth,*1 Marie-Christin Pauly,†1 Christian Schneider,‡ Annette Wittmer,‡ Sven Möllers,† Máté Döbrössy,† Christian Winkler,*2 and Guido Nikkhah†§2

*Department of Neurology, University Freiburg – Medical Center, Freiburg im Breisgau, Germany
†Stereotactic and Functional Neurosurgery, Department of General Neurosurgery, University Freiburg – Medical Center, Freiburg im Breisgau, Germany
‡Institute of Medical Microbiology and Hygiene, University of Freiburg, Freiburg im Breisgau, Germany
§Department of Neurosurgery, University Clinic Erlangen, Erlangen, Germany

Restorative cell therapy concepts in neurodegenerative diseases are aimed at replacing lost neurons. Despite advances in research on pluripotent stem cells, fetal tissue from routine elective abortions is still regarded as the only safe cell source. Progenitor cells isolated from distinct first-trimester fetal CNS regions have already been used in clinical trials and will be used again in a new multicenter trial funded by the European Union (TRANSEURO). Bacterial contamination of human fetal tissue poses a potential risk of causing infections in the brain of the recipient. Thus, effective methods of microbial decontamination and validation of these methods are required prior to approval of a neurorestorative cell therapy trial. We have developed a protocol consisting of subsequent washing steps at different stages of tissue processing. Efficacy of microbial decontamination was assessed on rat embryonic tissue incubated with high concentrations of defined microbe solutions including representative bacterial and fungal species. Experimental microbial contamination was reduced by several log ranks. Subsequently, we have analyzed the spectrum of microbial contamination and the effect of subsequent washing steps on aborted human fetal tissue; 47.7% of the samples taken during human fetal tissue processing were positive for a microbial contamination, but after washing, no sample exhibited bacterial growth. Our data suggest that human fetal tissue for neural repair can carry microbes of various species, highlighting the need for decontamination procedures. The decontamination protocol described in this report has been shown to be effective as no microbes could be detected at the end of the procedure.

Key words: Neural transplantation; Human fetus; Clinical application; Contamination; Sterility; Good manufacturing practice (GMP)

Received December 11, 2012; final acceptance April 2, 2013. Online prepub date: April 29, 2013.
1These authors provided equal contribution to this work.
2These authors provided equal contribution to this work.
Address correspondence to Guido Nikkhah, Department of Neurosurgery, University Clinic Erlangen, Schwabachanlage 6, 91054 Erlangen, Germany. Tel: +49-(0)9131-8544368; Fax: +49-(0)9131-8534569; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1009-1018, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X667484
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Strategies for Short-Term Storage of Hepatocytes for Repeated Clinical Infusions

Carl Jorns,* Roberto Gramignoli,† Mohammed Saliem,* Helen Zemack,* Lisa-Mari Mörk,* Bengt Isaksson,‡ Greg Nowak,* Bo-Göran Ericzon,* Stephen Strom,† and Ewa Ellis*

*Division of Transplantation Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institute, Karolinska University Hospital Huddinge, Stockholm, Sweden
†Division of Pathology, Department of Laboratory Medicine, Karolinska Institute, Karolinska University Hospital Huddinge, Stockholm, Sweden
‡Division of Surgery, Department of Clinical Science, Intervention and Technology (CLINTEC), Division of Transplantation Surgery, Karolinska Institute, Karolinska University Hospital Huddinge, Stockholm, Sweden

Hepatocyte transplantation is an upcoming treatment for patients with metabolic liver diseases. Repeated cell infusions over 1–2 days improve clinical outcome. Isolated hepatocytes are usually cold stored in preservation solutions between repeated infusions. However, during cold storage isolated hepatocytes undergo cell death. We investigated if tissue preservation and repeated isolations are better than storage of isolated hepatocytes when cold preserving human hepatocytes. Liver tissue obtained from liver surgery or organ donors was divided into two pieces. Hepatocytes were isolated by collagenase digestion. Hepatocytes were analyzed directly after isolation (fresh) or after storage for 48 h at 4°C in University of Wisconsin solution (UW cells). Liver tissue from the same donor was stored at 4°C in UW and hepatocytes were isolated after 48 h (UW tissue cells). Hepatocyte viability and function was evaluated by trypan blue exclusion, plating efficiency, ammonia metabolism, CYP 1A1/2, 2C9, 3A7, and 3A4 activities, phase II conjugation, and apoptosis evaluation by TUNEL assay and caspase-3/7 activities. Hepatocytes stored in UW showed a significantly lower viability compared to fresh cells or hepatocytes isolated from tissue stored for 48 h (54% vs. 71% vs. 79%). Plating efficiency was significantly decreased for cells stored in UW (40%) compared to fresh and UW tissue cells (63% vs. 55%). No significant differences between UW cells and UW tissue cells could be shown for CYP activities or ammonia metabolism. Hepatocytes stored in UW showed a strong increase in TUNEL-positive cells, whereas TUNEL staining in cold-stored liver tissue and hepatocytes isolated after 48 h was unchanged. This observation was confirmed by increased caspase-3/7 activities in UW cells. Although preservation of isolated hepatocytes in UW maintains function, cold storage of liver tissue and repeated hepatocyte isolations is superior to cold storage of isolated hepatocytes in preserving hepatocyte viability and function.

Key words: Hepatocyte transplantation; Cold storage; Preservation; Human hepatocytes; Apoptosis

Received November 12, 2012; final acceptance April 17, 2013. Online prepub date: April 29, 2013.
Address correspondence to Carl Jorns, M.D., Department of Clinical Science, Intervention and Technology (CLINTEC), Division of Transplantation Surgery, Karolinska Institute, Karolinska University Hospital Huddinge, Stockholm 14186, Sweden. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1019-1029, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X664568
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Improved Expression of Sirt1 on Thymic Epithelial Cells of SAMP10 After Intrabone Marrow–Bone Marrow Transplantation

Ming Li,* Ming Shi,* Nader G. Abraham,† and Susumu Ikehara*

*Department of Stem Cell Disorders, Kansai Medical University, Hirakata City, Osaka, Japan
†Department of Medicine, Marshall University Joan C. Edwards School of Medicine, Huntington, WV, USA

Aging is accompanied by various forms of immune dysfunction, leading to an increase in frequency of infections and the development of malignant tumors in mice and humans. Sirt1 has been implicated in processes as varied as metabolism, differentiation, cancer, and the stress response and aging. Senescence-accelerated mice prone 10 (SAMP10) show not only spontaneously occurring brain atrophy, with deficits in learning and memory, but also emotional disorders. We attempted in this study to clarify the deficits and found that the percentage of CD4/TNF-α T-cells in the spleen of 24-week-old (but not 6-week-old) SAMP10 to be significantly reduced. The thymus was significantly lighter, and the percentage of CD4+CD8+ cells was significantly lower in the 24-week-old SAMP10 than 6-week-old SAMP10. Microarray analyses indicated that genes related to transcription coactivator activity, growth factor activity, hormone activity, cytokine activity, receptor activity, and regulation of the immune system were downregulated in the thymus of 24-week-old SAMP10. Real-time PCR analysis showed that the expression of KGF, Aire, and Sirt1 was decreased on the thymic epithelial cells (TECs) of 24-week-old SAMP10. However, these parameters improved after the mice were treated with intrabone marrow–bone marrow transplantation. This is the first report of age-related changes in immune system dysfunction in 24-week-old SAMP10 and the first to show that dysfunction on the TECs of 24-week-old SAMP10 was modulated by allogeneic bone marrow cells.

Key words: Senescence-accelerated mice prone 10 (SAMP10); Aging; Sirt1; Thymic epithelial cells (TECs); Bone marrow transplantation

Received April 19, 2012; final acceptance February 7, 2013. Online prepub date: February 26, 2013.
Address correspondence to Susumu Ikehara, M.D., Ph.D., Department of Stem Cell Disorders, Kansai Medical University, Hirakata City, Osaka, 573-1010, Japan. Tel: +81-72-804-2450; Fax: +81-72-804-2454; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 23, pp. 1031-1044, 2014
0963-6897/14 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368913X665403
E-ISSN 1555-3892
Copyright © 2014 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

EpCAM Induction Functionally Links to the Wnt-Enhanced Cell Proliferation in Human Keratinocytes

Ching-I Shen,* Hsiu-Chin Lee,† Ying-Hsien Kao,‡ Chieh-Shan Wu,§ Po-Hung Chen,¶ Shinn-Zong Lin,#**†† Ping-Shan Lai,* and Hong-Lin Su†‡‡

*Department of Chemistry, Agricultural Biotechnology Center, National Chung-Hsing University, Taiwan
†Department of Life Sciences, Agricultural Biotechnology Center, National Chung-Hsing University, Taiwan
‡Department of Medical Research, E-DA Hospital, I-Shou University, Taiwan
§Department of Dermatology, Kaohsiung Veterans General Hospital, Taiwan
¶Chen Po-Hung Dermatologic Clinic, Kaohsiung, Taiwan
#Center for Neuropsychiatry, China Medical University and Hospital, Taiwan
**China Medical University Beigang Hospital, Taiwan
††Department of Immunology, China Medical University, Taiwan
‡‡Department of Physical Therapy, Graduate Institute of Rehabilitation Science, China Medical University, Taichung, Taiwan

Accelerating proliferation of primary keratinocytes benefits skin autografts for severely burned patients. Wnt signal, a conserved pathway controlling cell cycle and morphogenesis in embryo, also involves in cell proliferation and tumorigenesis in adult tissues. Here the effects of Wnt signal on the growth of human interfollicular keratinocytes were investigated. We demonstrated that recombinant Wnt3a significantly promoted the growth of primary keratinocytes at a low cell density. A well-characterized GSK-3β inhibitor, BIO, activated the Wnt signals and also enhanced the colony formation of keratinocytes dose dependently. Gene expression profile of the BIO-treated keratinocytes revealed the linkage of BIO with cell mitosis and indicated that epithelial cell adhesion molecule (EpCAM), a Wnt target gene, was significantly upregulated. Compared to the sorted EpCAM− keratinocytes, the EpCAM+ cells showed a higher proliferation rate and efficacy of colony formation. Inhibiting the EpCAM expression by shRNA attenuated the proliferation effect of BIO and the growth advantage of the EpCAM+ keratinocytes. These evidences emphasize the positive roles of canonical Wnt and EpCAM on the regulation of cell growth and self-renewal of human keratinocytes.

Key words: Wnt; Epithelial cell adhesion molecule (EpCAM); Proliferation; Keratinocyte; Epidermal progenitor cells

Received September 4, 2012; final acceptance March 21, 2013. Online prepub date: April 29, 2013.
Address correspondence to Hong-Lin Su, Department of Life Sciences, Agricultural Biotechnology Center, National Chung-Hsing University, 250 Kuo-Kuang Rd, Taichung 402, Taiwan. Tel: +886-4-22840416; Fax: +886-4-22854391, E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it