Cell Transplantation 21(11) Abstracts

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Cell Transplantation, Vol. 21, pp. 2325-2337, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X654957
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

G-CSF Stimulation and Coronary Reinfusion of Mobilized Circulating Mononuclear Proangiogenic Cells in Patients With Chronic Ischemic Heart Disease: Five-Year Results of the TOPCARE-G-CSF Trial

Joerg Honold,* Ulrich Fischer-Rasokat,* Ralf Lehmann,* David M. Leistner,* Florian H. Seeger,* Volker Schachinger,† Hans Martin,‡ Stefanie Dimmeler,§ Andreas M. Zeiher,* and Birgit Assmus*

*Division of Cardiology, Department of Medicine III, Goethe University Frankfurt, Frankfurt, Germany
†Department of Medicine I, Klinikum Fulda, Germany
‡Division of Hematology and Oncology, Department of Medicine II, Goethe University Frankfurt, Frankfurt, Germany
§Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University Frankfurt, Frankfurt, Germany

Prognosis of patients with heart failure remains poor despite improved conventional and interventional treatment regimens. The improvement of neovascularization and repair processes by administration of bone marrow-derived cells modestly improved the recovery after acute myocardial infarction. However, circulating patient-derived cells are reduced in number and function particularly in chronic heart failure. Therefore, we tested the hypothesis whether the mobilization of circulating mononuclear proangiogenic cells (CPCs) by G-CSF may overcome some of these limitations. In the present pilot study, 32 patients with at least 3-month-old myocardial infarction were randomized to G-CSF alone (G-CSF group) or intracoronary infusion of G-CSFmobilized and cultured CPCs into the infarct-related artery (G-CSF/CPC group). Primary endpoint of the study was safety. Efficacy parameters included serial assessment of LV function, NT-proBNP levels, and cardiopulmonary exercise testing. G-CSF effectively mobilized circulating CD34+CD45+ cells after 5 days in all patients (408 ± 64%) without serious adverse events. At 3 months, NYHA class and global LV function did not show significant improvements in both treatment groups (G-CSF: DLVEF 1.6 ± 2.4%; p = 0.10; G-CSF/CPC: DLVEF 1.4 ± 4.1%; p = 0.16). In contrast, target area contractility improved significantly in the G-CSF/CPC group. During 5-year follow-up, one patient died after rehospitalization for worsening heart failure. Eleven patients underwent further revascularization procedures. NT-proBNP levels, cardiopulmonary exercise capacity, and NYHA class remained stable in both treatment groups. The results from our pilot trial indicate that administration of G-CSF alone or G-CSF-mobilized and cultured CPCs can be performed safely in patients with chronic ischemic heart disease. However, only minor effects on LV function, NT-proBNP levels, and NYHA classification were observed during follow-up, suggesting that the enhancement of CPCs by G-CSF alone does not substantially improve intracoronary cell therapy effects in patients with chronic ischemic heart failure.

Key words: Heart failure; Ischemic heart disease; Granulocyte colony-stimulating factor (G-CSF); Cell therapy; Mononuclear cells

Received February 28, 2011; final acceptance February 7, 2012. Online prepub date: September 7, 2012.
Address correspondence to Birgit Assmus, M.D., Division of Cardiology, Department of Medicine III, Goethe University Frankfurt, Theodor Stern Kai 7 - 60590 Frankfurt, Germany. Tel: +49-69-6301-7461; Fax: +49-69-6301-7643; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2339-2350, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X655000
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Posttransplant Cellular Immune Reactivity Against Donor Antigen Correlates With Clinical Islet Transplantation Outcome: Towards a Better Posttransplant Monitoring

Stéphanie Lacotte,* Sophie Borot,* Sylvie Ferrari-Lacraz,† Jean Villard,† Sandrine Demuylder-Mischler,* Graziano Oldani,* Philippe Morel,* Gilles Mentha,* Thierry Berney,* and Christian Toso*

*Department of Surgery, Geneva University Hospitals, Faculty of Medicine, University of Geneva, Geneva, Switzerland
†Transplant Immunology Unit, Geneva University Hospitals, Faculty of Medicine, University of Geneva, Geneva, Switzerland

The aim of the present study was to assess the efficiency of cell-based immune assays in the detection of alloreactivity after islet transplantation and to correlate these results with clinical outcome. Mixed lymphocyte cultures were performed with peripheral blood mononuclear cells from recipients (n = 14), donors, or third party. The immune reactivity was assessed by the release of IFN-g (ELISpot), cell proliferation (FACS analysis for Ki67), and cytokine quantification (Bioplex). Islet function correlated with the number of IFN-γ-secreting cells following incubation with donor cells (p = 0.007, r = –0.50), but not with third party cells (p = 0.61). Similarly, a high number of donor-specific proliferating cells was associated with a low islet function (p = 0.006, r = −0.51). Proliferating cells were mainly CD3+CD4+ lymphocytes and CD3CD56+ natural killer cells (with low levels of CD3+CD8+ lymphocytes). Patients with low islet function had increased levels of CD4+Ki67+ cells (p ≤ 0.0001), while no difference was observed in CD8+Ki67+ and CD56+Ki67+ cells. IFN-γ, IL-5, and IL-17 levels were increased in patients with low islet function, but IL-10 levels tended to be lower. IFN- γ-ELISpot, proliferation, and cytokines were similarly accurate in predicting clinical outcome (AUC = 0.77 ± 0.088, 0.85 ± 0.084, and 0.88 ± 0.074, respectively). Cellular immune reactivity against donor cells correlates with posttransplant islet function. The tested assays have the potential to be of substantial help in the management of islet graft recipients and deserve prospective validation.

Key words: Monitoring; Diabetes; Islet; Transplantation; Cytokine; ELISpot

Received August 5, 2011; final acceptance February 5, 2012. Online prepub date: September 7, 2012.
Address correspondence to Christian Toso, Transplantation Unit, Geneva University Hospitals, rue Gabrielle-Perret-Gentil 4, 1211 Genève 14, Switzerland. Tel: +41 22 372 33 11; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2351-2362, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636957
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparative Study of Transplantation of Hepatocytes at Various Differentiation Stages Into Mice With Lethal Liver Damage

Ryo Kamimura,* Takamichi Ishii,* Naoya Sasaki,* Masatoshi Kajiwara,* Takafumi Machimoto,* Michiko Saito,† Kenji Kohno,† Hirofumi Suemori,‡ Norio Nakatsuji,§ Iwao Ikai,*¶ Kentaro Yasuchika,* and Shinji Uemoto*

*Department of Surgery, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto, Japan
†Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan
‡Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan
§Institute for Integrated Cell Material Sciences and Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan
¶Department of Surgery, National Hospital Organization Kyoto Medical Center, Fushimi-ku, Kyoto, Japan

Hepatocyte transplantation utilizing induced pluripotent stem cells (iPSCs) or embryonic stem cells (ESCs) has been expected to provide an alternative to liver transplantation. However, it remains uncertain precisely which cell type is the best suited for cell transplantation. In particular, it is unclear whether mature hepatocytes, which have sufficient liver function, or immature hepatic progenitor cells, which have a higher proliferative capacity, will provide a better outcome. The main objective of this study was to investigate the therapeutic efficacy of the transplantation of hepatocytes at various differentiation stages. We utilized transgenic mice that expressed diphtheria toxin (DT) receptors under the control of an albumin enhancer/promoter. ESC-derived endodermal cells, fetal hepatocytes, and adult hepatocytes were transplanted into these mice with experimentally induced lethal acute liver injury caused by DT administration. The transplanted cells were marked by enhanced green fluorescent protein. We evaluated their effects on survival. At 35 days after transplantation, the survival rate of the adult hepatocyte-transplanted group (8/20, 40%) was significantly improved in comparison to that of the sham-operated group (2/25, 8%), the fetal hepatocyte-transplanted group (1/20, 5%), and the ESC-derived endodermal cell-transplanted group (0/21, 0%). The adult hepatocytes proliferated in the recipient livers and replaced a large part of their parenchyma. The transplantation of adult hepatocytes for acute liver failure significantly improved the survival rate in comparison to that of transplantation of immature cells, thus suggesting that ESCs and iPSCs should be differentiated into mature hepatocytes before cell transplantation for acute liver failure.

Key words: Cell transplantation; Embryonic stem cell; Hepatocyte differentiation; Toxin receptor-mediated conditional cell knockout (TRECK)

Received March 23, 2011; final acceptance November 8, 2011. Online prepub date: April 2, 2011.
Address correspondence to Kentaro Yasuchika, M.D., Ph.D., Department of Surgery, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. Tel:+81-75-751-3242; Fax: +81-75-751-4263; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2363-2375, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638856
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Granulocyte Colony-Stimulating Factor and Interleukin-1β Are Important Cytokines in Repair of the Cirrhotic Liver After Bone Marrow Cell Infusion: Comparison of Humans and Model Mice

Yuko Mizunaga,* Shuji Terai,* Naoki Yamamoto,* Koichi Uchida,* Takahiro Yamasaki,* Hiroshi Nishina,† Yusuke Fujita,‡ Koh Shinoda,§ Yoshihiko Hamamoto,‡ and Isao Sakaida*

*Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan
†Department of Developmental and Regenerative Biology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan
‡Department of Computer Science and Systems Engineering, Faculty of Engineering, Yamaguchi University, Yamaguchi, Japan
§Department of Neuroanatomy and Neuroscience, Yamaguchi University School of Medicine, Yamaguchi, Japan

We previously described the effectiveness of autologous bone marrow cell infusion (ABMi) therapy for patients with liver cirrhosis (LC). We analyzed chronological changes in 19 serum cytokines as well as levels of specific cytokines in patients after ABMi therapy and in a mouse model of cirrhosis generated using green fluorescent protein (GFP)/carbon tetrachloride (CCl4). We measured expression profiles of cytokines in serum samples collected from 13 patients before and at 1 day and 1 week after ABMi. Child–Pugh scores significantly improved in all of these patients. To analyze the meaning of early cytokine change, we infused GFP-positive bone marrow cells (BMCs) into mice with CCl4-induced LC and obtained serum and tissue samples at 1 day and as well as at 1, 2, 3, and 4 weeks later. We compared chronological changes in serum cytokine expression in humans and in the model mice at 1 day and 1 week after BMC infusion. Among 19 cytokine, both granulocyte colony-stimulating factor (G-CSF) and interleukin-1β (IL-1β) in serum was found to show the same chronological change pattern between human and mice model. Next, we examined changes in cytokine expression in cirrhosis liver before and at 1, 2, 3, and 4 weeks after BMC infusion. Both G-CSF and IL-1β were undetectable in the liver tissues before and at 1 week after BMC infusion but increased at 2 weeks and continued until 4 weeks after infusion. The infused BMCs induced an early decrease of both G-CSF and IL-1β in serum and an increase in the model mice with LC. These dynamic cytokine changes might be important to repair liver cirrhosis after BMC infusion.

Key words: Autologous bone marrow infusion (ABMi); Cytokine; Liver regeneration; Cell therapy; Chronological change

Received August 23, 2010; final acceptance January 15, 2012. Online prepub date: April 10, 2012.
Address correspondence to Shuji Terai, M.D., Ph.D., Associate Professor, Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Minami Kogushi 1-1-1, Ube, Yamaguchi 755-8505, Japan. Tel: +81-836-22-2240; Fax: +81-836-22-2303; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2377-2395, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638892
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Achilles Tendon Regeneration Can Be Improved by Amniotic Epithelial Cell Allotransplantation

B. Barboni,*1 V. Russo,*1 V. Curini,*1 A. Mauro,*1 A. Martelli,* A. Muttini,*1 N. Bernabò,* L. Valbonetti,*1 M. Marchisio,†‡1 O. Di Giacinto,* P. Berardinelli,* and M. Mattioli*1

*Department of Comparative Biomedical Science, University of Teramo, Teramo, Italy
†Department of Medicine and Aging Science (D.M.S.I.), University “G. d’Annunzio” of Chieti-Pescara, Chieti, Italy
‡Center for Ageing Sciences (Ce.S.I.), “Università G. d’Annunzio” Foundation, Chieti, Italy

Amniotic epithelial cells (AECs) are ideal seed cells for tissue regeneration, but no research has yet been reported on their tendon regeneration potential. This study investigated the efficiency of AEC allotransplantation for tendon healing, as well as the mechanism involved. To this aim ovine AECs, characterized by specific surface and stemness markers (CD14, CD31, CD45, CD49f, CD29, CD166, OCT4, SOX2, NANOG, TERT), were allotransplanted into experimentally induced tissue defects in sheep Achilles tendon. In situ tissue repair revealed that AEC-treated tendons had much better structural and mechanical recoveries than control ones during the early phase of healing. Immunohistochemical and biochemical analyses indicated that extracellular matrix remodeling was more rapid and that immature collagen fibers were completely replaced by mature ones in 28 days. Moreover, spatial–temporal analysis of cellularity, proliferation index, vascular area, and leukocyte infiltration revealed that AECs induced a specific centripetal healing process that first started in the tissue closer to the healthy portion of the tendons, where AECs rapidly migrated to then progress through the core of the lesion. This peculiar healing evolution could have been induced by the growth factor stimulatory influence (TGF-β1 and VEGF) and/or by the host progenitor cells recruitment, but also as the consequence of a direct tenogenic AEC differentiation resulting in the regeneration of new tendon matrix. These findings demonstrate that AECs can support tendon regeneration, and their effects may be used to develop future strategies to treat tendon disease characterized by a poor clinical outcome in veterinary medicine.

Key words: Achilles tendon; Amniotic epithelial cells (AECs); Tissue regeneration; Cellular therapy

Received August 4, 2011; final acceptance February 1, 2012. Online prepub date: April 10, 2012.
1StemTeCh Group, Italy.
Address correspondence to Valentina Russo, Piazza A. Moro, 45 64100 Teramo, Italy. Tel: +390861266861; Fax: +390861266860; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2397-2405, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638865
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Chondrogenesis of Adipose Stem Cells in a Porous Polymer Scaffold: Influence of the Pore Size

Gun-II Im,* Ji-Yun Ko,* and Jin Ho Lee†

*Department of Orthopaedics, Dongguk University Ilsan Hospital, South Korea
†Department of Advanced Materials, Hannam University, Daejeon, South Korea

This study examined how the difference in pore size of porous scaffolds affected the in vitro chondrogenic differentiation of seeded adipose stem cells (ASCs) and the in vivo cartilage repair of ASC/scaffold construct. ASCs were isolated from 18 rabbits and seeded in a porous poly (e-caprolactone) (PCL) scaffold with different pore sizes (100, 200, 400 mm). The ASCs underwent in vitro chondrogenic induction under TGF-β2 and BMP-7 for 21 days before analysis. The ASC/scaffold construct was also implanted on the osteochondral defect created on the distal femur of the same rabbits, and the quality of cartilage regeneration was analyzed after 8 weeks. At day 21, the ASCs proliferated and spread on the surface of the scaffolds with a pore size 100 and 200 mm, whereas there were many lumps of conglomerated ASCs on those with a pore size of 400 μm. The DNA content was significantly lower in the scaffold with a pore size of 400 μm than in that with a pore size of 100 or 200 μm. Proteoglycan production was significantly greater in the scaffold with a pore size of 400 and 200 μm than in that with a pore size of 100 μm. The chondrogenic marker gene expression including SOX9 and COL2A1 was greatest in the scaffold with a pore size of 400 μm followed by 200 μm. Immunofluorescent imaging showed that, while SOX9 was localized to nucleus, type II collagen was observed on the cytoplasm and secreted matrix around the cells most abundantly in the scaffold with a pore size of 400 μm followed by 200 μm. The gross and histological findings from the osteochondral defects showed that the cartilage repair was better in the scaffold with a pore size of 400 and 200 μm than in that with a pore size of 100 μm.

Key words: Adipose stem cells (ASCs); Chondrogenesis; Cartilage regeneration; Pore size; Porous scaffold

Received August 30, 2011; final acceptance January 15, 2012. Online prepub date: April 10, 2012.
Address correspondence to Gun-II Im, Department of Orthopaedics, Dongguk University Ilsan Hospital, 814 Siksa-Dong, Goyang, 411-773, South Korea. Tel: +82 31 961 7314; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2407-2424, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637055
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Losartan Improves Adipose Tissue-Derived Stem Cell Niche by Inhibiting Transforming Growth Factor-β and Fibrosis in Skeletal Muscle Injury

Jin-Kyu Park,*1 Mi-Ran Ki,*1 Eun-Mi Lee,*† Ah-Young Kim,*† Sang-Young You,*† Seon-Young Han,*† Eun-Joo Lee,*† Il-Hwa Hong,* Soon-Hak Kwon,‡ Seong-Jin Kim,§ Thomas A. Rando,¶ and Kyu-Shik Jeong*†

*College of Veterinary Medicine, Kyungpook National University, Daegu, South Korea
†Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, South Korea
‡Department of Pediatrics, School of Medicine, Kyungpook National University, Daegu, South Korea
§CHA Cancer Institute, CHA University, Seoul, South Korea
¶Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA, USA

Recently, adipose tissue-derived stem cells (ASCs) were emerged as an alternative, abundant, and easily accessible source of stem cell therapy. Previous studies revealed losartan (an angiotensin II type I receptor blocker) treatment promoted the healing of skeletal muscle by attenuation of the TGF-β signaling pathway, which inhibits muscle differentiation. Therefore, we hypothesized that a combined therapy using ASCs and losartan might dramatically improve the muscle remodeling after muscle injury. To determine the combined effect of losartan with ASC transplantation, we created a muscle laceration mouse model. EGFP-labeled ASCs were locally transplanted to the injured gastrocnemius muscle after muscle laceration. The dramatic muscle regeneration and the remarkably inhibited muscular fibrosis were observed by combined treatment. Transplanted ASCs fused with the injured or differentiating myofibers. Myotube formation was also enhanced by ASC+ satellite coculture and losartan treatment. Thus, the present study indicated that ASC transplantation effect for skeletal muscle injury can be dramatically improved by losartan treatment inducing better niche.

Key words: Adipose tissue-derived stem cells (ASCs); Angiotensin II; Fibrosis; Losartan; Muscle laceration; Transforming growth factor-β (TGF-β)

Received August 26, 2011; final acceptance January 8, 2012. Online prepub date: April 10, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Kyu-Shik Jeong, D.V.M., Ph.D., Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, South Korea. Tel: +82-53-950-5975; Fax: +82-53-950-5955; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2425-2439, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X639008
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Adipose-Derived Stem Cells Improve Renal Function in a Mouse Model of IgA Nephropathy

Young Youl Hyun,*1 In Ok Kim,†‡1 Mi Hyung Kim,† Deok Hwa Nam,* Mi Hwa Lee,* Jung Eun Kim,* Hye Kyoung Song,* Jin Joo Cha,* Young Sun Kang,* Ji Eun Lee,§ Hyun Wook Kim,§ Jee Young Han,¶ and Dae Ryong Cha*

*Department of Internal Medicine, Korea University Ansan Hospital, Ansan, South Korea
†Anterogen Co., Ltd., Seoul, South Korea
‡Department of Bioinspired Science, Division of Life and Pharmaceutical Sciences, College of Pharmacy, Ewha Woman’s University, Seoul, South Korea
§Department of Internal Medicine, Wonkwang University, Gunpo, South Korea
¶Department of Pathology, Inha University Hospital, Incheon, South Korea

T-cell dysregulation plays an important role in the pathogenesis of immunoglobulin A nephropathy (IgAN). Adipose-derived stem cells (ASCs) have been reported to be able to prevent tissue damage through immunemodulating effects. To evaluate the effects of ASCs in high IgA ddY (HIGA) mice, ASCs were isolated from HIGA mice with different stages of IgAN before and after disease onset. ASCs were injected at a dose of 5 × 106 cells/kg body weight through the tail vein every 2 weeks for 3 months. Although the administered ASCs were rarely detected in the glomeruli, 24-h proteinuria was markedly decreased in all ASC-treated groups. Although glomerular deposition of IgA was not significantly different among groups, mesangial proliferation and glomerulosclerosis were dramatically decreased in most ASC treatment groups. In addition, levels of fibrotic and inflammatory molecules were markedly decreased by ASC treatment. Interestingly, ASC therapy significantly decreased Th1 cytokine activity in the kidney and caused a shift to Th2 responses in spleen T-cells as determined by FACS analysis. Furthermore, conditioned media from ASCs abrogated aggregated IgA-induced Th1 cytokine production in cultured HIGA mesangial cells. These results suggest that the beneficial effects of ASC treatment in IgAN occur via paracrine mechanisms that modulate the Th1/Th2 cytokine balance. ASCs are therefore a promising new therapeutic agent for the treatment of IgAN.

Key words: Adipose-derived stem cells (ASCs); HIGA mice; Th1 cytokine; Glomerulosclerosis; Proteinuria

Received August 26, 2011; final acceptance January 19, 2012. Online prepub date: April 17, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Dae Ryong Cha, Department of Internal Medicine, Korea University Ansan-Hospital, 516 Kojan-Dong, Ansan City, Kyungki-Do 425-020, South Korea. Tel: +82-031-412-4886; Fax: +82-031-412-5574; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2441-2454, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637064
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Adipose Tissue-Derived Mesenchymal Cells Support Skin Reepithelialization Through Secretion of KGF-1 and PDGF-BB: Comparison With Dermal Fibroblasts

Vassilia-Ismini Alexaki,* Despoina Simantiraki,* Marianna Panayiotopoulou,* Olga Rasouli,† Maria Venihaki,† Ourania Castana,‡ Dimitrios Alexakis,‡ Marilena Kampa,* Efstathios N. Stathopoulos,§ and Elias Castanas*

*Department of Experimental Endocrinology, School of Medicine, University of Crete, Heraklion, Greece
†Department of Clinical Chemistry, School of Medicine, University of Crete, Heraklion, Greece
‡Plastic and Reconstructive Surgery Department, Evaggelismos General Hospital, Athens, Greece
§Department of Pathology, School of Medicine, University of Crete, Heraklion, Greece

Epidermal organization and homeostasis are regulated by mesenchymal influences through paracrine actions. Until today, dermal fibroblasts (DFs) are used in the “dermal” layer to support keratinocyte growth in vitro in dermal and skin substitutes. In the present work, we used human adipose tissue-derived mesenchymal cells (ADMCs) as a support of keratinocyte growth in vitro (in monolayer culture and in 3D skin cell culture models) and in vivo (mouse wound healing models) and compared our findings with those obtained using dermal fibroblasts. ADMCs induce reepithelialization during wound healing more efficiently than DFs, by enhancing keratinocyte proliferation through cell cycle progression, and migration. This effect is mediated (at least partially) by a paracrine action of KGF-1 and PDGF-BB, which are more prominently expressed in ADMCs than in DFs. Furthermore, replacement of DFs by ADMCs in the dermal compartment of organotypic skin cultures leads to an artificial epidermis resembling to that of normal skin, concerning the general histology, although with a higher expression of cytokeratins 5 and 19. In Rag1 knockout mice, ADMCs induced a more rapid reepithelialization and a more effective wound healing, compared to dermal fibroblasts. In conclusion, we provide evidence that ADMCs can serve as supportive cells for primary keratinocyte cultures. In addition, because of their abundance and the great cell yield achieved during ADMC isolation, they represent an interesting cell source, with potential aspects for clinical use.

Key words: Adipose-derived mesenchymal cells (ADMCs); Dermal fibroblasts (DFs); Primary keratinocytes; Organotypic skin cultures; Wound healing

Received April 6, 2011; final acceptance November 1, 2011. Online prepub date: April 10, 2012.
Address correspondence to Dr. Elias Castanas, Laboratory of Experimental Endocrinology, School of Medicine, University of Crete, P.O. Box 2208, Heraklion, 71003, Greece. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2455-2469, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637037
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Environmental Biomechanics Substantiated by Defined Pillar Micropatterns Govern Behavior of Human Mesenchymal Stem Cells

S. Proksch,* T. Steinberg,† S. Schulz,† S. Sauerbier,‡ E. Hellwig,* and P. Tomakidi†

*Department of Operative Dentistry and Periodontology, Dental School and Hospital, University Freiburg Medical Center, Freiburg, Germany
†Department of Oral Biotechnology, Dental School and Hospital, University Freiburg Medical Center, Freiburg, Germany
‡Department of Oral and Maxillofacial Surgery, Dental School and Hospital, University Freiburg Medical Center, Freiburg, Germany

While evidence on the impact of the biomechanical environment elasticity on human mesenchymal stem cell (hMSC) behavior is growing, the aspect of micropatterning is still poorly understood. Thus, the present study aimed at investigating the influence of defined environmental micropatterning on hMSC behavior. Following characterization, hMSCs were grown on defined pillar micropatterns of 5, 7, 9, and 11 μm. With respect to cell behavior, primary hMSC adhesion was detected by indirect immunofluorescence (iIF) for paxillin, vinculin, integrin αV, and actin, while proliferation was visualized by histone H3. Morphogenesis was monitored by scanning electron microscopy and the expression of stem cell-specific biomarkers by real-time PCR. Favoritism of primary adhesion of hMSCs on pillar tops occurred at smaller pillar micropatterns, concomitant with cell flattening. While vinculin, integrin αV, and paxillin appeared initially more cytoplasmic, high pillar micropatterns favored a progressive redistribution with polarization to cell tension sites and at cell borders. Accomplishment of morphogenesis at day 3 revealed establishment of fully rotund cell somata at 5 μm, while hMSCs appeared progressively elongated at rising micropatterns. The hMSC proliferation capacity was influenced by pillar micropatterns and gene expression analysis of stem cell- and differentiation-associated biomarkers disclosed clear modulation by distinct pillar micropatterns. In response to environmental biomechanics, our results show that hMSC behavior is governed by pillar micropatterning. In turn, these findings may form the basis to prospectively direct lineage specificity of hMSCs in a customized fashion.

Key words: Human mesenchymal stem cells (hMSCs); Environmental micropattern; Biomechanics; Pillar array

Received September 22, 2011; final acceptance January 26, 2012. Online prepub date: April 2, 2012.
Address correspondence to S. Proksch, Department of Operative Dentistry and Periodontology, Dental School and Hospital, University Freiburg Medical Center, Hugstetter Str. 55, 79106 Freiburg, Germany. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2471-2486, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638874
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Differentiation of Chromaffin Progenitor Cells to Dopaminergic Neurons

Vladimir Vukicevic,*1 Janine Schmid,*1 Andreas Hermann,†‡ Sven Lange,* Nan Qin,§ Linda Gebauer,* Kuei-Fang Chung,* Ursula Ravens,¶ Graeme Eisenhofer,§ Alexander Storch,†‡# Marius Ader,‡ Stefan R. Bornstein,*‡ and Monika Ehrhart-Bornstein*‡

*Molecular Endocrinology, Medical Clinic III, University Clinic Dresden, Dresden University of Technology, Dresden, Germany
†Division of Neurodegenerative Diseases, Department of Neurology, Dresden University of Technology, Dresden, Germany
‡Center for Regenerative Therapies Dresden, Dresden University of Technology, Dresden, Germany
§Departments of Clinical Chemistry and Laboratory Medicine, University of Dresden, Dresden University of Technology, Dresden, Germany
¶Institute of Toxicology and Pharmacology, Medical Faculty Dresden, Dresden University of Technology, Dresden, Germany
#DZNE, German Centre for Neurodegenerative Diseases, Research Site Dresden, Dresden, Germany

The differentiation of dopamine-producing neurons from chromaffin progenitors might represent a new valuable source for replacement therapies in Parkinson’s disease. However, characterization of their differentiation potential is an important prerequisite for efficient engraftment. Based on our previous studies on isolation and characterization of chromaffin progenitors from adult adrenals, this study investigates their potential to produce dopaminergic neurons and means to enhance their dopaminergic differentiation. Chromaffin progenitors grown in sphere culture showed an increased expression of nestin and Mash1, indicating an increase of the progenitor subset. Proneurogenic culture conditions induced the differentiation into neurons positive for neural markers β-III-tubulin, MAP2, and TH accompanied by a decrease of Mash1 and nestin. Furthermore, Notch2 expression decreased concomitantly with a downregulation of downstream effectors Hes1 and Hes5 responsible for self-renewal and proliferation maintenance of progenitor cells. Chromaffin progenitor-derived neurons secreted dopamine upon stimulation by potassium. Strikingly, treatment of differentiating cells with retinoic and ascorbic acid resulted in a twofold increase of dopamine secretion while norepinephrine and epinephrine were decreased. Initiation of dopamine synthesis and neural maturation is controlled by Pitx3 and Nurr1. Both Pitx3 and Nurr1 were identified in differentiating chromaffin progenitors. Along with the gained dopaminergic function, electrophysiology revealed features of mature neurons, such as sodium channels and the capability to fire multiple action potentials. In summary, this study elucidates the capacity of chromaffin progenitor cells to generate functional dopaminergic neurons, indicating their potential use in cell replacement therapies.

Key words: Chromaffin progenitors; Neural differentiation; Parkinson’s disease; Ascorbic acid; Retinoic acid; Transplantation

Received July 5, 2011; final acceptance January 25, 2012. Online prepub date: April 10, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Monika Ehrhart-Bornstein, Ph.D., Molecular Endocrinology, Medical Clinic III, University Clinic Dresden, Fetscherstr, 74, 01307 Dresden, Germany. Tel: +49 351 458 6130; Fax: +49 351 458 7334; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2487-2496, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638964
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Neural Stem Cells Genetically Modified to Express Human Nerve Growth Factor (NGF) Gene Restore Cognition in the Mouse With Ibotenic Acid-Induced Cognitive Dysfunction

Hong J. Lee,*† In J. Lim,‡ Seung W. Park,§ Yun B. Kim,¶ Yong Ko,# and Seung U. Kim*†

*Medical Research Institute, Chung-Ang University College of Medicine, Seoul, South Korea
†Division of Neurology, UBC Hospital, University of British Columbia, Vancouver, Canada
‡Department of Physiology, Chung-Ang University College of Medicine, Seoul, South Korea
§Department of Neurosurgery, Chung-Ang University College of Medicine, Seoul, South Korea
Chungbuk National University School of Veterinary Medicine, Cheongju, South Korea
#Korea University College of Life Sciences and Biotechnology, Seoul, South Korea

Alzheimer’s disease (AD) is characterized by degeneration and loss of neurons and synapses throughout the brain, causing the progressive decline in cognitive function leading to dementia. No effective treatment is currently available. Nerve growth factor (NGF) therapy has been proposed as a potential treatment of preventing degeneration of basal forebrain cholinergic neurons in AD. In a previous study, AD patient’s own fibroblasts genetically modified to produce NGF were transplanted directly into the brain and protected cholinergic neurons from degeneration and improved cognitive function in AD patients. In the present study, human neural stem cells (NSCs) are used in place of fibroblasts to deliver NGF in ibotenic acid-induced learning-deficit rats. Intrahippocampal injection of ibotenic acid caused severe neuronal loss, resulting in learning and memory deficit. NGF protein released by F3.NGF human NSCs in culture medium is 10-fold over the control F3 naive NSCs at 1.2 μg/106 cells/day. Overexpression of NGF in F3.NGF cells induced improved survival of NSCs from cytotoxic agents H2O2, Aβ, or ibotenic acid in vitro. Intrahippocampal transplantation of F3.NGF cells was found to express NGF and fully improved the learning and memory function of ibotenic acid-challenged animals. Transplanted F3.NGF cells were found all over the brain and differentiated into neurons and astrocytes. The present study demonstrates that human NSCs overexpressing NGF improve cognitive function of learning-deficit model mice.

Key words: Learning and memory; Alzheimer’s disease (AD); Ibotenic acid; Human neural stem cell (NSC); Nerve growth factor (NGF); Gene therapy; Hippocampus

Received June 22, 2011; final acceptance February 23, 2012. Online prepub date: April 20, 2012.
Address correspondence to Seung U. Kim, M.D., Ph.D., Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, BC V6T 2B5, Canada. Tel: +1-604-822-7145; Fax: +1-604-822-7897; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Yong Ko, Ph.D., College of Life Sciences and Biotechnology, Korea University, Seoul 136-701, South Korea. Tel: 82-2-3290-3054; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2497-2515, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X640457
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Long-Lasting Paracrine Effects of Human Cord Blood Cells on Damaged Neocortex in an Animal Model of Cerebral Palsy

Sang-Hun Bae,*† Tae-Ho Kong,*† Hyun-Seob Lee,*† Kyung-Sul Kim, *† Kwan Soo Hong,‡ Michael Chopp,§¶ Myung-Seo Kang,# and Jisook Moon*†

*College of Life Science, Department of Applied Bioscience, CHA University, Seoul, South Korea
†General Research Institute, Gangnam CHA General Hospital, Seoul, South Korea
‡Division of MR Research Korea Basic Science Institute, Cheongwon, South Korea
§Department of Neurology, Henry Ford Hospital, Detroit, MI, USA
¶Department of Physics, Oakland University, Rochester, MI, USA
#Department of Diagnostic Tests, Kangnam CHA Hospital, Seoul, South Korea

Neonatal asphyxia is an important contributor to cerebral palsy (CP), for which there is no effective treatment to date. The administration of human cord blood cells (hUCBCs) is emerging as a therapeutic strategy for the treatment of neurological disorders. However, there are few studies on the application of hUCBCs to the treatment of neonatal ischemia as a model of CP. Experiments and behavioral tests (mainly motor tests) performed on neonatal hypoxia/ischemia have been limited to short-term effects of hUCBCs, but mechanisms of action have not been investigated. We performed a study on the use of hUCBCs in a rat model of neonatal hypoxia/ischemia and investigated the underlying mechanism for therapeutic benefits of hUCBC treatment. hUCBCs were intravenously transplanted into a rat model of neonatal hypoxia ischemia. hUCBCs increased microglia temporarily in the periventricular striatum in the early phase of disease, protected mature neurons in the neocortex from injury, paved the way for the near-normalization of brain damage in the subventricular zone (SVZ), and, in consequence, significantly improved performance in a battery of behavioral tests compared to the vehicle-treated group. Although the transplanted cells were rarely observed in the brain 3 weeks after transplantation, the effects of the improved behavioral functions persisted. Our preclinical findings suggest that the long-lasting positive influence of hUCBCs is derived from paracrine effects of hUCBCs that stimulate recovery in the injured brain and protect against further brain damage.

Key words: Cerebral palsy (CP); Human umbilical cord blood stem cells (hUCBCs); Inflammatory cytokine; Paracrine factors; Chemokines

Received June 27, 2011; final acceptance December 20, 2011. Online prepub date: April 17, 2012.
Address correspondence to Jisook Moon, Department of Applied Bioscience, CHA University, 606-16 Yeoksam-1 dong, Gangnam-gu, Seoul, South Korea. Tel: +82-2-3468-3196; Fax: +82-2-538-4102; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp.2517-2521, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X637425
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Definitive Setup of Clinical Scale Procedure for Ex Vivo Expansion of Cord Blood Hematopoietic Cells for Transplantation

Pascale Duchez,*† Jean Chevaleyre,*† Marija Vlaski,*† Bernard Dazey,* Noel Milpied,‡ Jean-Michel Boiron,*† and Zoran Ivanovic*†

*Etablissement Français du Sang Aquitaine-Limousin, Bordeaux, France
†CNRS/Université Segalen UMR 5164, Bordeaux, France
‡CHU de Bordeaux/Université Segalen, Bordeaux, France

We recently developed a clinical grade ex vivo cord blood expansion procedure enabling a massive amplification of hematopoietic progenitors without any loss of stem cell potential. This procedure, based on day 14 liquid cultures of cord blood CD34+ cells, in medium Macopharma HP01 and in the presence of stem cell factor (SCF; 100 ng/ml), fms-related tyrosine kinase 3-ligand (Flt-3L; 100 ng/ml), megakaryocyte growth and developmental factor (MGDF; 100 ng/ml), and granulocyte colony-stimulating factor (G-CSF; 10 ng/ml) had to be modified due to the commercially unavailability of clinical grade MGDF molecule. So MGDF was replaced by thrombopoietin (TPO) in fivefold lower dose (20 ng/ml), and culture time was reduced to 12 days. That way, a mean expansion fold of 400, 80, and 150 was obtained for total cells, CD34+ cells, and colony-forming cells (CFCs), respectively. This amplification was associated with a slight enhancing effect on stem cells [Scid repopulating cells (SRCs)]. These are the ultimate preclinical modifications of a clinical grade expansion protocol, which is already employed in an ongoing clinical trial.

Key words: Ex vivo; Hematopoietic; Progenitors; Expansion; Cord blood (CB)

Received June 15, 2011; final acceptance January 5, 2012. Online prepub date: March 28, 2012.
Address correspondence to Zoran Ivanovic, Etablissement Français du Sang Aquitaine Limousin, BP 24, 5 Place Amélie Raba Léon, 33035 Bordeaux Cedex, France. Tel: +33 5 56 90 75 50; Fax: +33 5 56 90 75 51; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2523-2530, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653165
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Comparison of Cardiomyogenic Potential Among Human ESC and iPSC Lines

Ana Sepac,* Karim Si-Tayeb,† Filip Sedlic,* Sara Barrett,* Scott Canfield,* Stephen A. Duncan,† Zeljko J. Bosnjak,* and John W. Lough†

*Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, WI, USA
†Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI, USA

We recently reported that, following induction of clumps of pluripotent H1 human embryonic stem cells (hESCs) with activin-A and Bmp4 in defined medium for 5 days, widespread differentiation of rhythmically contracting cardiomyocytes occurs within 3–4 weeks. In this study, the same approach was used to assess whether human induced pluripotent stem cells (hiPSCs), which may theoretically provide an unlimited source of patientmatched cells for transplantation therapy, can similarly undergo cardiomyocyte differentiation. Differentiation of four pluripotent cell lines (H1 and H9 hESCs and C2a and C6a hiPSCs) was compared in parallel by monitoring rhythmic contraction, morphologic differentiation, and expression of cardiomyogenic genes. Based on expression of the cardiomyogenic lineage markers MESP1, ISL1, and NKX2-5, all four cell lines were induced into the cardiomyogenic lineage. However, in contrast to the widespread appearance of striations and rhythmic contractility seen in H9 and especially in H1 hESCs, both hiPSC lines exhibited poor terminal differentiation. These findings suggest that refined modes of generating hiPSCs, as well as of inducing cardiomyogenesis in them, may be required to fulfill their potential as agents of cardiac regeneration.

Key words: Human embryonic stem cells (hESCs); Human induced pluripotent stem cells (hiPSCs); Directed cardiac differentiation; qPCR

Received August 4, 2011; final acceptance January 10, 2012. Online prepub date August 2, 2012.
Address correspondence to John W. Lough, Ph.D. Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI, USA. Tel: +1-414-955-8459; Fax: +1-414-955-6517; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2531-2535, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636975
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Collagenase Does Not Persist in Human Islets Following Isolation

Sarah E. Cross,* Stephen J. Hughes,* Anne Clark,† Derek W. R. Gray,* and Paul R. V. Johnson*

*Islet Transplant Research Group, Nuffield Department of Surgical Sciences, University of Oxford, John Radcliffe Hospital, Oxford, UK
†Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, UK

Optimal human islet isolation requires the delivery of bacterial collagenase to the pancreatic islet–exocrine interface. However, we have previously demonstrated the presence of collagenase within human islets immediately following intraductal collagenase administration. This potentially has significant implications for patient safety. The present study aimed to determine if collagenase becomes internalized into islets during the isolation procedure and if it remains within the islet postisolation. Islet samples were taken at various stages throughout 14 clinical human islet isolations: during digest collection, following University of Wisconsin solution incubation, immediately postisolation, and after 24 h of culture. Samples were embedded in agar, cryosectioned, and then assessed by immunolabeling for collagenase and insulin. Immunoreactivity for collagenase was not observed in isolated islets in any preparation. Collagenase labeling was detected in one sample taken at the digest collection phase in one islet preparation only. No collagenase-specific labeling was seen in islets sampled at any of the other time points in any of the 14 islet preparations. Collagenase that enters islets during intraductal administration is washed out of the islets during the collection phase of the isolation process and thus does not remain in islets after isolation. This observation alleviates some of the important safety concerns that collagenase remains within islet grafts.

Key words: Islet isolation; Islet transplantation; Collagenase

Received August 10, 2011; final acceptance January 10, 2012. Online prepub date April 2, 2012.
Address correspondence to Dr. Sarah E. Cross, Islet Transplant Research Group, Nuffield Department of Surgical Sciences, Level 6, John Radcliffe Hospital, Oxford, OX3 9DU, UK. Tel: +44 (0)1865 857507; Fax: +44 (0)1865 768876; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it