Cell Transplantation 21(10) Abstracts

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Cell Transplantation, Vol. 21, pp. 2099-2110, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636786
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Reversal of Diabetes in Mice With a Bioengineered Islet Implant Incorporating a Type I Collagen Hydrogel and Sustained Release of Vascular Endothelial Growth Factor

Robert B. Vernon,* Anton Preisinger,* Michel D. Gooden,* Leonard A. D’Amico,* Betty B. Yue,* Paul L. Bollyky,* Christian S. Kuhr,†Thomas R. Hefty,† Gerald T. Nepom,* and John A. Gebe*

*Benaroya Research Institute at Virginia Mason, Seattle, WA, USA
†Virginia Mason Medical Center, Seattle, WA, USA

We have developed a bioengineered implant (BI) to evaluate strategies to promote graft survival and function in models of islet transplantation in mice. The BI, sized for implantation within a fold of intestinal mesentery, consists of a disk-shaped, polyvinyl alcohol sponge infused with a type I collagen hydrogel that contains dispersed donor islets. To promote islet vascularization, the BI incorporates a spherical alginate hydrogel for sustained release of vascular endothelial growth factor (VEGF). BIs that contained 450–500 islets from syngeneic (C57Bl/6) donors and 20 ng of VEGF reversed streptozotocin (STZ)-induced diabetes in 100% of mice (8/8), whereas BIs that contained an equivalent number of islets, but which lacked VEGF, reversed STZ-induced diabetes in only 62.5% of mice (5/8). Between these “+VEGF” and “–VEGF” groups, the time to achieve normoglycemia (8–18 days after implantation) did not differ statistically; however, transitory, postoperative hypoglycemia was markedly reduced in the +VEGF group relative to the –VEGF group. Notably, none of the mice that achieved normoglycemia in these two groups required exogenous insulin therapy once the BIs began to fully regulate levels of blood glucose. Moreover, the transplanted mice responded to glucose challenge in a near-normal manner, as compared to the responses of healthy, nondiabetic (control) mice that had not received STZ. In future studies, the BIs described here will serve as platforms to evaluate the capability of immunomodulatory compounds, delivered locally within the BI, to prevent or reverse diabetes in the setting of autoimmune (type 1) diabetes.

Key words: Diabetes; Islet; Bioengineered implant (BI); Collagen; Vascular endothelial growth factor (VEGF); Mouse

Received May 11, 2011; final acceptance December 2, 2011. Online prepub date: March 28, 2012.
Address correspondence to John A. Gebe, Ph.D., Benaroya Research Institute at Virginia Mason, 1201 Ninth Ave., Seattle, WA 98101, USA. Tel: +1-206-287-5612; Fax: +1-206-342-6581; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2111-2118, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637399
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Activation of Peroxisome Proliferator-Activated Receptor γ Prolongs Islet Allograft Survival

Hongjun Wang,*† Hongju Wu,‡ Fredy Rocuts,† Zhuoying Gu,† Fritz H. Bach,† and Leo E. Otterbein†

*Department of Surgery, Medical University of South Carolina, Charleston, SC, USA
†Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA
‡Department of Obstetrics and Gynecology, The University of Alabama at Birmingham, Birmingham, AL, USA

Exposing donor mice to carbon monoxide (CO) protects transplanted islet allografts from immune rejection after transplantation (referred as the “donor” effect). In an attempt to understand the mechanisms of the donor effect of CO, we found that donor treatment with CO upregulates expression of peroxisome proliferator-activated receptor γ (PPARγ), a transcriptional regulator, in isolated islets. In this study, we evaluated whether PPARγ contributes to the survival and function of transplanted islets and whether PPARγ mediates the protective effect of CO in a major mismatch islet allogeneic transplantation model. BALB/c (H-2d) islets in which PPARγ activity was induced by its agonists, 15-deoxy-Δ12-14-prostaglandin J2 (15d-PGJ2) or troglitazone were transplanted into C57BL/6 (H-2b) recipients that had been rendered diabetic by streptozotocin (STZ). Blood glucose levels of recipients were monitored to determine the function of transplanted islets. Our data indicated that PPARγ activation in islets led to a high percentage of BALB/c islets survived long-term in C57BL/6 recipients. Activation of PPARγ in the donor suppresses expressions of proinflammatory cytokines including tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase (iNOS) in transplanted islets. Blocking PPARγ activity by its antagonist, GW9662, abrogated the donor effect of CO in vivo and in vitro. Our data demonstrate that PPARγ plays a critical role in the survival and function of transplanted islets after transplantation in the recipient. The protective effects of CO are at least in part mediated by PPARγ.

Key words: Peroxisome proliferator-activated receptor γ (PPARγ); Carbon monoxide; Islet transplantation; Allograft survival

Received May 9, 2011; final acceptance December 2, 2011. Online prepub date: March 28, 2012.
Address correspondence to Hongjun Wang, Ph.D., Department of Surgery, Medical University of South Carolina, Charleston, SC 29425, USA. Tel: +1-843-792-1800; Fax: +1-843-792-3315; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2119-2129, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X638955
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Increased β-Cell Replication and β-Cell Mass Regeneration in Syngeneically Transplanted Rat Islets Overexpressing Insulin-Like Growth Factor II

Elisabet Estil·les,*† Noèlia Téllez,*† Jessica Escoriza,‡ and Eduard Montanya*†‡

*Laboratory of Diabetes and Experimental Endocrinology, Department of Clinical Sciences, IDIBELL-University of Barcelona, Barcelona, Spain
†CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Barcelona, Spain
‡Endocrine Unit (15), IDIBELL-Hospital Universitari Bellvitge, Barcelona, Spain

Insulin-like growth factor II (IGF2) is a growth-promoting peptide that increases β-cell proliferation and survival. The aim of the study was to determine the effect of IGF2 overexpression on β-cell mass in transplanted islets. Islets infected with adenovirus encoding for IGF2 (Ad-IGF2 group), for luciferase (Ad-Luc control group), or with uninfected islets (control group) were syngeneically transplanted to streptozotocin-diabetic Lewis rats. Eight hundred islets, a minimal mass model to restore normoglycemia, or 500 islets, a clearly insufficient mass, were transplanted. Rats transplanted with 800 Ad-IGF2 islets showed a better metabolic evolution than control groups. As expected, rats transplanted with 500 Ad-IGF2 or control islets maintained similar hyperglycemia throughout the study, ensuring comparable metabolic conditions among both groups. β-Cell replication was higher in Ad-IGF2 group than in control group on days 3 [1.45% (IQR: 0.26) vs. 0.58% (IQR: 0.18), p = 0.006], 10 [1.58% (IQR: 1.40) vs. 0.90% (IQR: 0.61), p = 0.035], and 28 [1.35% (IQR: 0.35) vs. 0.64% (IQR: 0.28), p = 0.004] after transplantation. β-Cell mass was similarly reduced on day 3 after transplantation in Ad-IGF2 and control group [0.36 mg (IQR: 0.26) vs. 0.38 mg (IQR: 0.19)], it increased on day 10, and on day 28 it was higher in Ad-IGF2 than in control group [0.63 mg (IQR: 0.38) vs. 0.42 mg (IQR: 0.31), p = 0.008]. Apoptosis was similarly increased in Ad-IGF2 and control islets after transplantation. No differences in insulin secretion were found between Ad-IGF2 and uninfected control islets. In summary, IGF2 overexpression in transplanted islets increased β-cell replication, induced the regeneration of the transplanted β-cell mass, and had a beneficial effect on the metabolic outcome reducing the β-cell mass needed to achieve normoglycemia.

Key words: Insulin-like growth factor II (IGF2); Islet transplantation; Pancreatic β-cell; β-Cell mass; β-Cell proliferation

Received March 15, 2010; final acceptance December 13, 2011. Online prepub date: April 10, 2012.
Address correspondence to Eduard Montanya, Endocrine Unit (15), Hospital Universitari Bellvitge, Feixa Llarga, s/n, 08907 L’Hospitalet de Llobregat, Barcelona, Spain. Tel: +34-93-4037265; Fax: +34-93-2607561; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp.2131-2147, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636803
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Insulin Secreted From Insulinogenic Xenograft Restores Normoglycemia in Type 1 Diabetic Mice Without Immunosuppression

J. Kim,* S. Park,*† H. M. Kang,* C. W. Ahn,‡ H. C. Kwon,§ J. H. Song,¶ Y. J. Lee,¶ K. H. Lee,# H. Yang,† S. Y. Baek,** S. H. Yoo,** S. H. Kim,** and H. Kim†

*bcellbio, Inc., Seoul, South Korea
†Department of Biotechnology, College of Natural Sciences, Seoul Women’s University, Seoul, South Korea
‡Department of Internal Medicine, Yonsei University College of Medicine, Seoul, South Korea
§Mirae & Heemang OB/GYN Clinic, Seoul, South Korea
¶Medi-i Women’s Hospital, Seoul, South Korea
#Division of Cancer Research, Korea Institute of Radiological and Medical Sciences, Seoul, South Korea
**Korea Food and Drug Administration, Cheongwon-gun, Chungcheongbuk-do, South Korea

In the present study, we examined the therapeutic potential of human amnion-derived insulin-secreting cells for type 1 diabetes. Human amniotic mesenchymal stem cells (hAMs) were isolated from amnion and cultivated to differentiate into insulin-secreting cells in vitro. After culture in vitro, the differentiated cells (hAM-ISCs) were intensively stained with dithizone and secreted insulin and c-peptide in a high-glucose-dependent manner. They expressed mRNAs of pancreatic cell-related genes, including INS, PDX1, Nkx6-1, NEUROG3, ISL1, NEUROD1, GLUT1, GLUT2, PC1/3, PC2, GCK, PPY, SST, and GC, and were positive for human insulin and c-peptide. Transplantation of hAM-ISCs into the kidneys of mice with streptozotocin-induced diabetes restored body weight and normalized the blood glucose levels, which lasted for 210 days. Only human insulin and c-peptide were detected in the blood of normalized mice after 2 months of transplantation, but little mouse insulin and c-peptide. Removal of graft-bearing kidneys from these mice resulted in causing hyperglycemia again. Human cell-specific gene, hAlu, and human pancreatic cell-specific genes, insulin, PDX1, GLUT1, GLP1R, Nkx6-1, NEUROD1, and NEUROG3, were detected in the graft-bearing kidneys. Colocalization of human insulin and human nuclei antigen was also observed. These results demonstrate that hAMs could differentiate into functional insulin-secreting cells in vitro, and human insulin secreted from hAM-ISCs following transplantation into type 1 diabetic mice could normalize hyperglycemia, overcoming immune rejection for a long period.

Key words: Human amnion mesenchymal stem cells (hAMs); Insulin-secreting cells (ISCs); Type 1 diabetes; Insulin; β-Cells; Immunocompetent

Received March 22, 2011; final acceptance December 23, 2011. Online prepub date: April 2, 2012.
Address correspondence to Haekwon Kim, Department of Biotechnology, College of Natural Sciences, Seoul Women’s University, Seoul 139-774, South Korea. Tel: +82-2-970-5665; Fax: +82-2-974-2473; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2149-2157, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636911
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Noninvasive Monitoring of Immunosuppressive Drug Efficacy to Prevent Rejection of Intracerebral Glial Precursor Allografts

Michael Gorelik,*†1 Miroslaw Janowski,*†‡§1 Chulani Galpoththawela,*† Robert Rifkin,¶ Michael Levy,¶ Barbara Lukomska,‡ Douglas A. Kerr,†† Jeff W. M. Bulte,*†#** and Piotr Walczak*†

*Russell H. Morgan Department of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
†Cellular Imaging Section, Vascular Biology Program, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
‡NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
§Department of Neurosurgery, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland
¶Department of Neurology, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
#Department of Chemical & Biomolecular Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
**Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
††Neurodegeneration and Translational Neurology, Biogen Idec, Cambridge, MA, USA

The development of cell-based therapies opens up new avenues for treating a myriad of diseases of the central nervous system (CNS). While significant effort is being directed toward development of patient-specific, autologous transplantable cells, at present, the majority of cell transplantation studies performed clinically utilize allografts. In this context, the issue of graft rejection and immunoprotection is of key importance. In this study, we transplanted mouse glial-restricted progenitors into immunodeficient, immunocompetent, and immunosuppressed mice and monitored their survival noninvasively using bioluminescence imaging (BLI). With the use of serial BLI, we evaluated both the prevalence and dynamics of cell rejection. We demonstrate that allografts in immunocompetent mice were rejected at a rate of 69.2% (n = 13) indicating that graft tolerance is possible even without immunosuppression. Immunosuppression using a combination of rapamycin and FK506 or cyclosporine failed to fully protect the grafts. FK506 and rapamycin treatment resulted in a slight improvement of immunoprotection (22.2% rejected, n = 9) compared to cyclosporin A (55.6% rejected, n = 9); however, the difference was not significant. Notably, immunohistochemistry revealed leukocytes infiltrating the graft area in both rejecting and nonrejecting immunocompetent animals, but not in immunodeficient animals. The induction of an inflammatory process, even in surviving allografts, has implications for their long-term survival and functionality.

Key words: Glial restricted precursors; Transplantation; Bioluminescence; Immunosuppression; Rejection; Allograft

Received May 25, 2011; final acceptance December 1, 2011. Online prepub date: April 10, 2012.
1These authors provided equal contribution to this work.
Address correspondence to Piotr Walczak, Russell H. Morgan Department of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, 733 N Broadway 649 BRB, Baltimore, MD 21205, USA. Tel: +1 4432875614; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2159-2169, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X627543
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Suppression of Astrocyte Lineage in Adult Hippocampal Progenitor Cells Expressing Hippocampal Cholinergic Neurostimulating Peptide Precursor in an In Vivo Ischemic Model

Takanari Toyoda,* Noriyuki Matsukawa,* Takafumi Sagisaka,* Norihiko Uematsu,* Tetsuko Kanamori,* Daisuke Kato,* Masayuki Mizuno,* Hiroaki Wake,* Hideki Hida,† Cesario V. Borlongan,‡ and Kosei Ojika*

*Department of Neurology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan
†Department of Neurophysiology and Brain Science, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan
‡Department of Neurosurgery and Brain Repair, University of South Florida Morsani College of Medicine, Tampa, FL, USA

Hippocampal cholinergic neurostimulating peptide (HCNP) is known to promote differentiation of septohippocampal cholinergic neurons. The HCNP precursor protein (HCNP-pp) may play several roles, for example, as an ATP-binding protein, a Raf kinase inhibitor protein, and a phosphatidylethanolamine-binding protein, as well as a precursor for HCNP. This study therefore aimed to elucidate the involvement of HCNP-pp in specific neural lineages after stroke using a hypoxic–ischemic (HI) rat model of brain ischemia. The specific neural lineages in the hippocampus were investigated 14 days after ischemia. Some bromodeoxyuridine (BrdU)+ neural progenitor cells in the hippocampus of hypoxic, HI, or sham-operated rats expressed HCNP-pp. Almost half of the BrdU+/HCNP-pp+ cells also expressed the oligodendrocyte lineage marker 2′,3′-cyclic nucleotide 3′-phosphodiesterase, whereas only a few BrdU+/HCNP-pp+ cells in the hippocampus in HI brains expressed the neuronal lineage marker, doublecortin (DCX). Interestingly, no BrdU+/HCNP-pp+ progenitor cells in hypoxic, HI, or sham-operated brains expressed the astrocyte lineage marker, glial fibrillary acidic protein. Together with previous in vitro data, the results of this study suggest that the expression level of HCNP-pp regulates the differentiation of neural progenitor cells into specific neural lineages in the HI hippocampus, indicating that neural stem cell fate can be controlled via the HCNP-pp mediating pathway.

Key words: Hippocampal cholinergic neurostimulating peptide precursor protein; Adult rat hippocampal progenitor cell; Neural differentiation; Hypoxic–ischemic model

Received June 15, 2011; final acceptance November 29, 2011. Online prepub date: March 14, 2012.
Address correspondence to Noriyuki Matsukawa, M.D., Ph.D., Associate Professor, Department of Neurology and Neuroscience, Nagoya City University Graduate School of Medical Sciences, 1 Kawashumi, Mizuho-cho, Mizuho-ku, Nagoya, Aichi, Japan. Tel: +81-52-853-8094; Fax: +81-52-852-3590; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2171-2187, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X639035
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Production of Reactive Oxygen Species by Multipotent Stromal Cells/Mesenchymal Stem Cells Upon Exposure to Fas Ligand

Melanie Rodrigues,* Omari Turner,† Donna Stolz,‡ Linda G. Griffith,§ and Alan Wells*

*Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA
†Meharry Medical College, Nashville, TN, USA
‡Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA
§Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

Multipotent stromal cells (MSCs) can be differentiated into osteoblasts and chondrocytes, making these cells candidates to regenerate cranio-facial injuries and lesions in long bones. A major problem with cell replacement therapy, however, is the loss of transplanted MSCs at the site of graft. Reactive oxygen species (ROS) and nonspecific inflammation generated at the ischemic site have been hypothesized to lead to MSCs loss; studies in vitro show MSCs dying both in the presence of ROS or cytokines like FasL. We questioned whether MSCs themselves may be the source of these death inducers, specifically whether MSCs produce ROS under cytokine challenge. On treating MSCs with FasL, we observed increased ROS production within 2 h, leading to apoptotic death after 6 h of exposure to the cytokine. N-acetyl cysteine, an antioxidant, is able to protect MSCs from FasL-induced ROS production and subsequent ROS-dependent apoptosis, though the MSCs eventually succumb to ROS-independent death signaling. Epidermal growth factor (EGF), a cell survival factor, is able to protect cells from FasL-induced ROS production initially; however, the protective effect wanes with continued FasL exposure. In parallel, FasL induces upregulation of the uncoupling protein UCP2, the main uncoupling protein in MSCs, which is not abrogated by EGF; however, the production of ROS is followed by a delayed apoptotic cell death despite moderation by UCP2. FasL-induced ROS activates the stress-induced MAPK pathways JNK and p38MAPK as well as ERK, along with the activation of Bad, a proapoptotic protein, and suppression of survivin, an antiapoptotic protein; the latter two key modulators of the mitochondrial death pathway. FasL by itself also activates its canonical extrinsic death pathway noted by a time-dependent degradation of c-FLIP and activation of caspase 8. These data suggest that MSCs participate in their own demise due to nonspecific inflammation, holding implications for replacement therapies.

Key words: Reactive oxygen species (ROS); Multipotent stromal cells; Marrow stromal cells; Multipotent stem cells; Mesenchymal stem cells

Received March 24, 2011; final acceptance November 22, 2011. Online prepub date April 17, 2012.
Address correspondence to Alan Wells, S 713 Scaife Hall, 3550 Terrace St., Pittsburgh, PA 15261, USA. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2189-2200, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636821
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Effects of Donor Characteristics and Ex Vivo Expansion on Canine Mesenchymal Stem Cell Properties: Implications for MSC-Based Therapies

Susan W. Volk,*† Yanjian Wang,* and Kurt D. Hankenson†

*Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA
†Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA

Clinical trials utilizing bone marrow-derived mesenchymal stem cell (BM-MSC) therapies show promise for treating a variety of pathologic conditions. Paramount to optimization of such cell-based therapies is a thorough understanding of MSC biology. Despite the tremendous potential that exists for the clinical use of canine BM-MSCs in veterinary medicine, as well as in preclinical studies for human medicine, relatively little information exists regarding basic biological properties of the cells. In this study, we compared the importance of donor characteristics (age and harvest site) and ex vivo expansion on canine BM-MSC frequency (CFU-f) and differentiation potential. Advancing age was found to have a negative effect on CFU-f as well as osteogenic potential. Site of harvest was also found to have significant effects on MSC properties. MSCs obtained from the humerus were found at the lowest frequency and were least osteogenic compared to those harvested from the tibia, femur, and ilium. Osteogenic potential diminished significantly by the third passage. These results suggest important donor parameters and culture effects to consider in translational studies examining MSC-based regenerative medical strategies.

Key words: Mesenchymal stem cell (MSC); Regenerative medicine; Veterinary cell therapy; Canine; Tissue engineering; Aging

Received August 25, 2011; final acceptance December 21, 2011. Online prepub date: April 2, 2012.
Address correspondence to Susan W. Volk, V.M.D., Ph.D., University of Pennsylvania School of Veterinary Medicine, 312 Hill Pavilion, 380 S. University Ave., Philadelphia, PA 19104-4539, USA. Tel: +1-215-898-0635; Fax: +1-215-746-2295; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2201-2214, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368911X637362
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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A Novel Marker of Human Endometrial Mesenchymal Stem-Like Cells

Hirotaka Masuda,* Siti S. Anwar,* Hans-Jorg Buhring,† Jyothsna R. Rao,* and Caroline E. Gargett*

*The Ritchie Centre, Monash Institute of Medical Research, and Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria, Australia
†Department of Internal Medicine II, Divisions of Hematology, Oncology, Immunology, and Rheumatology, Laboratory for Stem Cell Research, University Clinic of Tubingen, Tubingen, Germany

Coexpression of CD140b (PDGFRβ) and CD146 has been used to isolate endometrial mesenchymal stem-like cells (eMSCs), which have a perivascular location. This study aims to evaluate a single marker for purifying eMSCs. Using an antibody panel with novel specificities, we screened human endometrial tissues and stromal cell suspensions by flow cytometry and immunohistochemistry to identify perivascular markers. Sorted subpopulations were examined for colony-forming unit (CFU), self-renewal, and differentiation assays for mesenchymal stem cell (MSC) function. We also transplanted sorted eMSCs under the kidney capsule of superimmunodeficient NSG mice. Magnetic bead selection was compared with flow cytometry sorting (flow sorting) using CFU assay. One novel marker (W5C5) was particularly effective in selecting eMSCs. W5C5+ cells comprise 4.2 ± 0.6% (n = 34) of endometrial stromal cells and reside predominantly in a perivascular location in both basal and functional layers of endometrium. The clonogenicity of W5C5+ cells is significantly greater than W5C5 and unselected cells. W5C5+ cells differentiated into adipocytes, osteocytes, chondrocytes, myocytes, and endothelial cells. W5C5+ cells produce endometrial stromal-like tissue in vivo. In terms of clonogenicity, magnetic bead-selected W5C5+ cells gave rise to significantly higher CFU numbers compared to flow-sorted W5C5+ cells. This study identified W5C5 as a single marker capable of purifying eMSCs possessing MSC properties and reconstituting endometrial stromal tissues in vivo. W5C5 enriches eMSCs to high purity and provides a simple protocol for their prospective isolation using magnetic bead selection rather than flow sorting. W5C5 selection may provide an alternate, readily available autologous source of MSC, obtainable with minimal morbidity using an office endometrial biopsy procedure for future cell-based therapies.

Key words: W5C5; Endometrium; Mesenchymal stem/stromal cells; Prospective isolation; Single maker; Tissue regeneration

Received June 27, 2011; final acceptance December 20, 2011. Online prepub date: March 27, 2012.
Address correspondence to Caroline E. Gargett, Ph.D., The Ritchie Centre, Monash Institute of Medical Research and Department of Obstetrics and Gynaecology, Monash University 27-31 Wright Street, P.O. Box 5418, Clayton, Victoria, 3168, Australia. Tel: +61 3 9902 4712; Fax: +61 3 9594 7439; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2215-2224, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653048
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Human Menstrual Blood-Derived Mesenchymal Cells as a Cell Source of Rapid and Efficient Nuclear Reprogramming

Deivid de Carvalho Rodrigues,* Karina Dutra Asensi,* Leandro Vairo,* Ricardo Luiz Azevedo-Pereira,* Rosane Silva,* Edson Rondinelli,*† Regina Coeli Goldenberg,* Antonio Carlos Campos de Carvalho,‡ and Turán Péter Ürményi*

*Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
†Departamento de Clínica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
‡Instituto Nacional de Cardiologia, Rio de Janeiro, Brazil

Induced pluripotent stem cells (iPSCs) were originally generated by forced ectopic expression of four transcription factors genes—OCT4, KLF4, SOX2, and c-MYC—in fibroblasts. However, the efficiency of iPSCs obtention is extremely low, and reprogramming takes about 20 days. We reasoned that adult cells showing basal expression of core embryonic stem (ES) cell regulator genes could be a better cell source for reprogramming. Menstrual blood-derived mesenchymal cells (MBMCs) are multipotent cells that show detectable levels of some of the core ES cells regulators. The aim of this study was to determine whether reprogramming efficiency could be increased by using MBMCs as a cell source to generate iPSCs. MBMCs were transduced with recombinant retroviruses expressing the coding regions of OCT4, SOX2, and KLF4 genes. Cells with high nucleus/cytoplasm ratio can be detected about 5 days of posttransduction, and colonies of typical ES-like cells begun to appear after 7 days. At day 15, colonies were picked up and expanded for characterization. Most of the clones were morphologically identical to ES cells and positive at the mRNA and protein levels for all pluripotency markers tested. The clones are capable of forming embryoid bodies and to differentiate in vitro into cells of the three germ cell layers. Our results show that the reprogramming was faster and with efficiency around 2–5%, even in the absence of ectopic expression of c-MYC. To date, this is the first study showing MBMCs as a cell source for nuclear reprogramming.

Key words: Induced pluripotent stem cells (iPSCs); Mesenchymal stromal cells; Reprogramming efficiency; Gene expression

Received April 14, 2011; final acceptance November 10, 2011. Online prepub date: July 5, 2012.
Address correspondence to Turán P. Ürményi, IBCCF, Bl. G, CCS, UFRJ, Cidade Universitária, Rio de Janeiro, RJ 21949-900, Brazil. Tel: +5521-2564-7364; Fax: +5521-2280-8193; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2225-2239, 2012
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DOI: http://dx.doi.org/10.3727/096368912X653020
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
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Improved Cell Survival and Paracrine Capacity of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells Promote Therapeutic Potential for Pulmonary Arterial Hypertension

Yuelin Zhang,* Songyan Liao,* Mo Yang,†‡ Xiaoting Liang,* Ming-Wai Poon,* Chee-Yin Wong,§ Junwen Wang,¶ Zhongjun Zhou,¶ Soon-Keng Cheong,§ Chuen-Neng Lee,# Hung-Fat Tse,*,** and Qizhou Lian*,**††

*Cardiology Division, Department of Medicine, University of Hong Kong, Hong Kong
†Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou, PR China
‡Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong
§MAKNA Cancer Research Institute, Kuala Lumpur, Malaysia
¶Department of Biochemistry, University of Hong Kong, Hong Kong
#Department of Surgery, National University Hospital, National University of Singapore, Singapore
**Research Centre of Heart, Brain, Hormone, and Healthy Aging, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong
††Eye Institute, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong

Although transplantation of adult bone marrow mesenchymal stem cells (BM-MSCs) holds promise in the treatment for pulmonary arterial hypertension (PAH), the poor survival and differentiation potential of adult BM-MSCs have limited their therapeutic efficiency. Here, we compared the therapeutic efficacy of human embryonic stem cell-derived MSCs (hESC-MSCs) with adult BM-MSCs for the treatment of PAH in an animal model. One week following monocrotaline (MCT)-induced PAH, mice were randomly assigned to receive phosphate-buffered saline (MCT group); 3.0 × 106 human BM-derived MSCs (BM-MSCs group) or 3.0 × 106 hESC-derived MSCs (hESC-MSCs group) via tail vein injection. At 3 weeks posttransplantation, the right ventricular systolic pressure (RVSP), degree of RV hypertrophy, and medial wall thickening of pulmonary arteries were lower, and pulmonary capillary density was higher in the hESC-MSC group as compared with BM-MSC and MCT groups (all p < 0.05). At 1 week posttransplantation, the number of engrafted MSCs in the lungs was found significantly higher in the hESC-MSC group than in the BM-MSC group (all p < 0.01). At 3 weeks posttransplantation, implanted BM-MSCs were undetectable whereas hESC-MSCs were not only engrafted in injured pulmonary arteries but had also undergone endothelial differentiation. In addition, protein profiling of hESC-MSC- and BM-MSC-conditioned medium revealed a differential paracrine capacity. Classification of these factors into bioprocesses revealed that secreted factors from hESC-MSCs were preferentially involved in early embryonic development and tissue differentiation, especially blood vessel morphogenesis. We concluded that improved cell survival and paracrine capacity of hESC-MSCs provide better therapeutic efficacy than BM-MSCs in the treatment for PAH.

Key words: Pulmonary arterial hypertension; Embryonic stem cell-derived mesenchymal stem cells

Received October 7, 2010; final acceptance October 10, 2011. Online prepub date: July 5, 2012.
Address correspondence to Qizhou Lian, M.D., Ph.D., or Hung-Fat Tse, M.D., Ph.D., Cardiology Division, Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Hong Kong. Tel: +852-21899752; Fax: +852-28162095; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it (Qizhou Lian), This e-mail address is being protected from spambots. You need JavaScript enabled to view it (Hung-Fat Tse)


Cell Transplantation, Vol. 21, pp. 2241-2255, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X639026
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adult-Derived Human Liver Progenitor Cells in Long-Term Culture Maintain Appropriate Gatekeeper Mechanisms Against Transformation

Isabelle Scheers,* Cedric Maerckx,* Dung Ngoc Khuu,* Sabrina Marcelle,† Anabelle Decottignies,‡ Mustapha Najimi,* and Etienne Sokal*

*Université Catholique de Louvain, Institut de Recherche Clinique et Expérimentale (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Brussels, Belgium
†Université Catholique de Louvain, Center for Human Genetics, Brussels, Belgium
‡Université Catholique de Louvain, de Duve Institute, Genetic and Epigenetic Alterations of Genomes Unit, Brussels, Belgium

The use of human liver progenitor cells in the development of clinical cell therapy depends on their constant availability and unaltered properties during culture. The present study investigates the effects of long-term in vitro culture on the specific characteristics of these cells and on their genetic stability. Adult-derived human liver progenitor cells (ADHLPCs) were isolated from 12 donors and cultured until senescence and cell death. Cells were analyzed at different time points for their phenotype stability and differentiation potential. In addition, growth characteristics, chromosomal karyotype, telomere maintenance mechanisms, and activity of cell cycle-related genes were studied. Finally, their in vivo tumorigenicity was investigated in a xenograft assay. The long-term culture of ADHLPCs revealed a variable proliferation capacity. Cells maintained their original phenotype and acquired hepatocyte-like metabolic functions after differentiation. Eight of the 12 cell populations grew fast (doubling time of 6.3 days) during a limited time period (mean, 116.2 days), and mainly presented normal cytogenetic features. The four other cell cultures presented an early decline in growth potential (doubling time of 28.6 days) and premature senescence. Chromosomal alterations were detected in three of four cultures at passage 6. Cytogenetic anomalies were not correlated with tumorigenic potential in vitro or in vivo, and expression of cell cycle-related genes was appropriately upregulated, inducing senescence. Although chromosomal anomalies may occur in long-term cell cultures, neither transformation nor alteration of their characteristics was noted during in vitro expansion. All ADHLPCs reached senescence and growth arrest. Presenescent ADHLPCs might therefore be considered as a suitable source for liver-based cell therapy.

Key words: Liver cell therapy; Progenitor cells; Cytogenetic instability; Cell senescence

Received January 31, 2011; final acceptance December 1, 2011. Online prepub date: April 17, 2012.
Address correspondence to Etienne Sokal, Université Catholique de Louvain, Cliniques Universitaires Saint Luc, Laboratory of Pediatric Hepatology and Cell Therapy, 52nd Avenue Mounier-Tour Vésale +3B-1200 Brussels, Belgium. Tel: +32-2-764-13-87; Fax: +32-2-764-89-09; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2257-2266, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637000
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Hyaluronan-Supplemented Buffers Preserve Adhesion Mechanisms Facilitating Cryopreservation of Human Hepatic Stem/Progenitor Cells

Rachael A. Turner,*† Gemma Mendel,† Eliane Wauthier,* Claire Barbier,*1 and Lola M. Reid*†‡

*Department of Cell Biology and Physiology, University of North Carolina School of Medicine, Chapel Hill, NC, USA
†Department of Biomedical Engineering, University of North Carolina School of Medicine, Chapel Hill, NC, USA
‡Program in Molecular Biology and Biotechnology, University of North Carolina School of Medicine, Chapel Hill, NC, USA

The supply of human hepatic stem cells (hHpSCs) and other hepatic progenitors has been constrained by the limited availability of liver tissues from surgical resections, the rejected organs from organ donation programs, and the need to use cells immediately. To facilitate accessibility to these precious tissue resources, we have established an effective method for serum-free cryopreservation of the cells, allowing them to be stockpiled and stored for use as an off-the-shelf product for experimental or clinical programs. The method involves use of buffers, some serum-free, designed for cryopreservation and further supplemented with hyaluronans (HA) that preserve adhesion mechanisms facilitating postthaw culturing of the cells and preservation of functions. Multiple cryopreservation buffers were found to yield high viabilities (80–90%) of cells on thawing of the progenitor cells. Serum-free CS10 supplemented with 0.05% hyaluronan proved the most effective, both in terms of viabilities of cells on thawing and in yielding cell attachment and formation of expanding colonies of cells that stably maintain the stem/progenitor cell phenotype. Buffers to which 0.05% or 0.1% HAs were added showed cells postthaw to be phenotypically stable as stem/progenitors, as well as having a high efficiency of attachment and expansion in culture. Success correlated with improved expression of adhesion molecules, particularly CD44, the hyaluronan receptor, E-cadherin, β4 integrin in hHpSCs, and β1 integrins in hepatoblasts. The improved methods in cryopreservation offer more efficient strategies for stem cell banking in both research and potential therapy applications.

Key words: Human hepatic stem cells (hHpSCs); Cryopreservation; Hyaluronans (HA); Regenerative medicine

Received July 19, 2011; final acceptance December 8, 2011. Online prepub date: April 2, 2012.
1Posthumous.
Address correspondence to Lola M. Reid, Department of Cell Biology and Physiology and Program in Molecular Biology and Biotechnology, University of North Carolina School of Medicine, Campus Box 7038, Glaxo Building Rooms 32–35, Chapel Hill, NC 27599, USA. Tel: +1-919-966-0347; Fax: +1-919-6112; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2267-2282, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X637505
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Clinical Outcome of Hepatocyte Transplantation in Four Pediatric Patients With Inherited Metabolic Diseases

Carmen Ribes-Koninckx,*† Eugenia Pareja Ibars,‡ Maria Angeles Calzado Agrasot,*† Ana Bonora-Centelles,† Begona Polo Miquel,* Juan Jose Vila Carbo,§ Ester Donat Aliaga,* Jose Mir Pallardo,‡ Maria Jose Gomez-Lechon,†¶#1 and Jose V. Castell†¶#**1

*Paediatric Gastroenterology and Hepatology Unit, University La Fe Hospital, Valencia, Spain
†Unit for Cell Therapy, University La Fe Hospital, Valencia, Spain
‡Liver Transplantation Unit, University La Fe Hospital, Valencia, Spain
§Paediatric Surgery, University La Fe Hospital, Valencia, Spain
¶Experimental Hepatology Unit, University La Fe Hospital, Valencia, Spain
#CIBERehd, FIS, Spain
**Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Valencia, Spain

Hepatocyte transplantation (HT) has become an effective therapy for patients with metabolic inborn errors. We report the clinical outcome of four children with metabolic inborn errors that underwent HT, describing the cell infusion protocol and the metabolic outcome of transplanted patients. Cryopreserved hepatocytes were used as this allows scheduling of treatments. Functional competence (viability, cell attachment, major cytochrome P450 and UDP-glucuronosyltransferase 1A1 activities, and urea synthesis) and microbiological safety of cell batches were assessed prior to clinical use. Four pediatric patients with liver metabolic diseases [ornithine transcarbamylase (OTC) deficiency, Crigler–Najjar (CNI) syndrome, glycogen storage disease Ia (GSD-Ia), and tyrosinemia type I (TYR-I)] underwent HT. Indication for HT was based on severity of disease, deterioration of quality of life, and benefits for the patients, with the ultimate goal to improve their clinical status whenever liver transplantation (LT) was not indicated or to bridge LT. Cells were infused into the portal vein while monitoring portal flow. The protocol included antibiotic prophylaxis and immunosuppressant therapy. After HT, analytical data on the disease were obtained. The OTC-deficient patient showed a sustained decrease in plasma ammonia levels and increased urea production after HT. Further cell infusions could not be administered given a fatal nosocomial fungus sepsis 2 weeks after the last HT. The CNI and GSD-Ia patients improved their clinical status after HT. They displayed reduced serum bilirubin levels (by ca. 50%) and absence of hypoglycaemic episodes, respectively. In both cases, the HT contributed to stabilize their clinical status as LT was not indicated. In the infant with TYR-I, HT stabilized temporarily the biochemical parameters, resulting in the amelioration of his clinical status while diagnosis of the disease was unequivocally confirmed by full gene sequencing. In this patient, HT served as a bridge therapy to LT.

Key words: Hepatocyte transplantation (HT); Human hepatocytes; Cryopreservation; Inherited metabolic diseases; Ornithine transcarbamylase (OTC) deficiency; Crigler–Najjar (CNI) syndrome; Glycogen storage disease Ia (GSD-Ia); Tyrosinemia type I (TYR-I); Pediatrics

Received February 9, 2010; final acceptance December 18, 2011. Online prepub date: April 10, 2012.
1Shared supervision.
Address correspondence to Maria Jose Gomez-Lechon, Unidad de Hepatologia Experimental, Instituto de Investigacion Sanitaria, University Hospital La Fe, Avda Campanar 21; E-46009, Valencia, Spain. E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2283-2297, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X653129
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparing Scaffold-Free and Fibrin-Based Adipose-Derived Stromal Cell Constructs for Adipose Tissue Engineering: An In Vitro and In Vivo Study

Femke Verseijden,* Sandra J. Posthumus-van Sluijs,* Johan W. van Neck,* Stefan O. P. Hofer,† Stevan E. R. Hovius,* and Gerjo J. V. M. van Osch‡§

*Department of Plastic and Reconstructive Surgery, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands
†Division of Plastic Surgery, University Health Network, University of Toronto, Toronto, Ontario, Canada
‡Department of Orthopedics, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands
§Department of Otorhinolaryngology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands

Success of adipose tissue engineering for soft tissue repair has been limited by insufficient adipogenic differentiation, an unfavorable host response, and insufficient vascularization. In this study, we examined how scaffold-free spheroid and fibrin-based environments impact these parameters in human adipose-derived stromal cell (ASC)-based adipose constructs. ASCs were differentiated in spheroids or fibrin-based constructs. After 7 days, conditioned medium was collected and spheroids/fibrin-based constructs were either harvested or implanted subcutaneously in athymic mice. Following 7 days of implantation, the number of blood vessels in fibrin-based constructs was significantly higher than in spheroids (93 ± 45 vs. 23 ± 11 vessels/mm2) and the inflammatory response to fibrin-based constructs was less severe. The reasons for these results were investigated further in vitro. We found that ASCs in fibrin-based constructs secreted significantly higher levels of the angiogenic factors VEGF and HGF and lower levels of the inflammatory cytokine IL-8. Furthermore, ASCs in fibrin-based constructs secreted significantly higher levels of leptin and showed a 2.5-fold upregulation of the adipogenic transcription factor PPARG and a fourfold to fivefold upregulation of the adipocyte-specific markers FABP4, perilipin, and leptin. These results indicate that fibrin-based ASC constructs are potentially more suitable for ASC-based adipose tissue reconstruction than scaffold-free spheroids.

Key words: Mesenchymal stem cell; Adipose tissue engineering; Fibrin; Cytokines; Inflammation; Angiogenesis

Received May 4, 2010; final acceptance December 5, 2011. Online prepub date: July 26, 2012.
Address correspondence to Femke Verseijden, Department of Plastic and Reconstructive Surgery, Erasmus MC, University Medical Center Rotterdam, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands. Tel: +31107043242; Fax: +317044685; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2299-2312, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636795
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Healthy Bone Marrow Cells Reduce Progression of Kidney Failure Better Than CKD Bone Marrow Cells in Rats With Established Chronic Kidney Disease

Arianne van Koppen,* Jaap A. Joles,* Lennart G. Bongartz,* Jens van den Brandt,† Holger M. Reichardt,† Roel Goldschmeding,‡ Tri Q. Nguyen,‡ and Marianne C. Verhaar*

*Department of Nephrology and Hypertension, University Medical Center Utrecht, Utrecht, The Netherlands
†Department of Cellular and Molecular Immunology, University of Göttingen Medical School, Göttingen, Germany
‡Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands

Chronic kidney disease (CKD) is a major health care problem. New interventions to slow or prevent disease progression are urgently needed. We studied functional and structural effects of infusion of healthy and CKD bone marrow cells (BMCs) in a rat model of established CKD. CKD was induced by 5/6 nephrectomy (SNX) in Lewis rats, and disease progression was accelerated with l-NNA and 6% NaCl diet. Six weeks after SNX, CKD rats received healthy eGFP+ BMCs, CKD eGFP+ BMCs, or vehicle by single renal artery injection. Healthy BMCs were functionally effective 6 weeks after administration: glomerular filtration rate (GFR; inulin clearance) (0.48 ± 0.16 vs. 0.26 ± 0.14 ml/min/100 g) and effective renal plasma flow (RPF; PAH clearance) (1.6 ± 0.40 vs. 1.0 ± 0.62 ml/min/100 g) were higher in healthy BMC- versus vehicle-treated rats (both p < 0.05). Systolic blood pressure (SBP) and proteinuria were lower 5 weeks after treatment with healthy BMCs versus vehicle (SBP, 151 ± 13 vs. 186 ± 25 mmHg; proteinuria, 33 ± 20 vs. 59 ± 39 mg/day, both p < 0.05). Glomerular capillary density was increased, and less sclerosis was detected after healthy BMCs (both p < 0.05). Tubulointerstitial inflammation was also decreased after healthy BMCs. eGFP+ cells were present in the glomeruli and peritubular capillaries of the remnant kidney in all BMC-treated rats. CKD BMCs also reduced SBP, proteinuria, glomerulosclerosis, and tubular atrophy versus vehicle in CKD rats. However, CKD BMC therapy was not functionally effective versus vehicle [GFR: 0.28 ± 0.09 vs. 0.26 ± 0.16 ml/min/100 g (NS), RPF: 1.15 ± 0.36 vs. 0.78 ± 0.44 ml/min/100 g (NS)], and failed to decrease tubulointerstitial inflammation and fibrosis. Single intrarenal injection of healthy BMCs in rats with established CKD slowed progression of the disease, associated with increased glomerular capillary density and less sclerosis, whereas injection of CKD BMCs was less effective.

Key words: Bone marrow cells (BMCs); Kidney; Renal hemodynamics; Endothelium

Received December 22, 2010; final acceptance November 30, 2011. Online prepub date: March 28, 2012.
Address correspondence to Marianne C. Verhaar, M.D., Ph.D., Department of Nephrology and Hypertension, F03.223, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands. Tel: +31 88 7557329; Fax: +31 30 2518328; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation, Vol. 21, pp. 2313-2324, 2012
0963-6897/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/096368912X636867
E-ISSN 1555-3892
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Dermal Papilla Cells Serially Cultured With Wnt-10b Sustain Their Hair Follicle Induction Activity After Transplantation Into Nude Mice

Yukiteru Ouji,*† Shigeaki Ishizaka,† and Masahide Yoshikawa*†

*Department of Pathogen, Infection, and Immunity, Nara Medical University, Kashihara, Nara, Japan
†Program in Tissue Engineering, Department of Parasitology, Nara Medical University, Kashihara, Nara, Japan

Dermal papilla (DP) cells are associated with the development of hair follicles (HFs) and regulation of the hair cycle. However, primary DP cells prepared from cultured HFs are known to lose their ability to induce HF after culturing in standard media, for example, fibroblast growth conditions. We explored a new culture condition by which DP cells maintained their HF induction ability. The addition of Wnt-10b to the first culture of primary DP cells promoted their proliferation and maintained their Wnt responsiveness and HF induction ability. Furthermore, DP cells in Wnt-10b-containing medium sustained those characteristics after 10 passages (100 days), which encompassed the entire experimental period. These results suggest that Wnt-10b plays a pivotal role in proliferation and maintenance of DP cells in vitro.

Key words: Dermal papilla (DP); Transplantation; Wnt signaling; Wnt-10b; Hair growth; Hair reconstitution

Received August 7, 2011; final acceptance December 1, 2011. Online prepub date: April 2, 2012.
Address correspondence to Yukiteru Ouji or Masahide Yoshikawa, Department of Pathogen, Infection, and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara, 634-8521, Japan. Tel: +81-744-29-8847; Fax: +81-744-29-8847; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it (Y.O.) or This e-mail address is being protected from spambots. You need JavaScript enabled to view it (M.Y.)