Cell Transplantation 19(11) Abstracts

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Cell Transplantation, Vol. 19, pp. 1359–1367, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X513964
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Liposomal Formulations of Thrombomodulin Increase Engraftment After Intraportal Islet Transplantation

Wanxing Cui,* Julianty Angsana,† Jing Wen,* and Elliot L. Chaikof*†‡

*Department of Surgery, Emory University, Atlanta, GA, USA
†Department of Biomedical Engineering, Emory University/Georgia Institute of Technology, Atlanta, GA, USA
‡School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA

Early destruction of donor islet grafts due to an instant blood-mediated inflammatory reaction (IBMIR) remains a major obstacle in islet transplantation. Thrombomodulin plays an important role in limiting coagulation and inflammatory events through a variety of effects. In this study, we investigated the ability of thrombomodulin (TM), when reconstituted as a liposomal formulation, to enhance early syngeneic islet engraftment by minimizing or abrogating the IBMIR. Administration of TM significantly improved early engraftment of syngeneic islets after intraportal transplantation in diabetic mice. In the absence of treatment, conversion to euglycemia was observed among 46.6% (7/15) of recipients. In contrast, administration of TM led to euglycemia in 93.3% (14/15) of recipients (p = 0.0142). Recipients that received TM exhibited a lower incidence of primary nonfunction and better glucose control over a 30-day period after transplantation. Fibrin deposition (p < 0.05), neutrophil infiltration (p < 0.05), expression of TNF-α and IL-β mRNA (p < 0.05), as well as NF-κB activity (p < 0.05) were significantly reduced in the liver of islet recipients having been treated with liposomal TM. These data demonstrate that TM significantly improves early syngeneic islet engraftment through effects that target both coagulation and inflammatory pathways.

Key words: Islet transplantation; Thrombosis; Inflammation; Engraftment; Thrombomodulin

Address correspondence to Elliot L. Chaikof, M.D., Ph.D., Emory University, 101 Woodruff Circle, Rm 5105, Atlanta, GA 30322, USA. Tel: (404) 727-8413; Fax: (404)-727-3660; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1369–1381, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X514288
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

The Mobilization and Effect of Endogenous Bone Marrow Progenitor Cells in Diabetic Wound Healing

Paolo Fiorina,†‡1 Giorgio Pietramaggiori,*§1 Saja S. Scherer,*§ Mollie Jurewicz,† Jasmine C. Mathews,* Andrea Vergani,†‡ Gebhard Thomas,¶ Elena Orsenigo,‡ Carlo Staudacher,‡ Stefano La Rosa,** Carlo Capella,** Adelaide Carothers,†† Hans-Gunter Zerwes,¶ Livio Luzi,# Reza Abdi,† and Dennis P. Orgill*

*Plastic Surgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA
†Transplantation Research Center, Division of Nephrology, Children’s Hospital and Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA
‡Department of Medicine & Surgery, San Raffaele Scientific Institute, Milan, Italy
§Department of Surgery, University of Geneva, Geneva, Switzerland
¶Autoimmunity, Transplantation and Inflammation and Global Discovery Chemistry, Novartis Institutes for Biomedical Research, Basel, Switzerland
#Facoltà di Scienze Motorie, Università di Milano, Milan, Italy
**Department of Pathology, Ospedale di Circolo and Department of Human Morphology, University of Insubria, Varese, Italy
††Department of Surgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

Diabetic patients suffer from impaired wound healing, characterized by only modest angiogenesis and cell proliferation. Stem cells may stimulate healing, but little is known about the kinetics of mobilization and function of bone marrow progenitor cells (BM-PCs) during diabetic wound repair. The objective of this study was to investigate the kinetics of BM-PC mobilization and their role during early diabetic wound repair in diabetic db/db mice. After wounding, circulating hematopoietic stem cells (Lin-c-Kit+Sca-1+) stably increased in the periphery and lymphoid tissue of db/db mice compared to unwounded controls. Peripheral endothelial progenitor cells (CD34+VEGFR+) were 2.5- and 3.5-fold increased on days 6 and 10 after wounding, respectively. Targeting the CXCR4–CXCL12 axis induced an increased release and engraftment of endogenous BM-PCs that was paralleled by an increased expression of CXCL12/SDF-1α in the wounds. Increased levels of peripheral and engrafted BM-PCs corresponded to stimulated angiogenesis and cell proliferation, while the addition of an agonist (GM-CSF) or an antagonist (ACK2) did not further modulate wound healing. Macroscopic histological correlations showed that increased levels of stem cells corresponded to higher levels of wound reepithelialization. After wounding, a natural release of endogenous BMPCs was shown in diabetic mice, but only low levels of these cells homed in the healing tissue. Higher levels of CXCL12/SDF-1α and circulating stem cells were required to enhance their engraftment and biological effects. Despite controversial data about the functional impairment of diabetic BM-PCs, in this model our data showed a residual capacity of these cells to trigger angiogenesis and cell proliferation.

Key words: Bone marrow progenitor cells (BM-PCs); Diabetes; Wound healing; Cell proliferation

1
These authors provided equal contribution to the work.
Address correspondence to Dennis P. Orgill, M.D., Ph.D., Division of Plastic Surgery, Brigham and Women’s Hospital/Harvard Medical School, 75 Francis Street, Boston, MA 02115, USA. Tel: (617) 732-5456; Fax: (617) 730-2855; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Paolo Fiorina, M.D., Ph.D., Transplantation Research Center, Division of Nephrology, Children’s Hospital/Harvard Medical School, 221 Longwood Avenue, Boston, MA 02115, USA. Tel: (617) 919-2624; Fax: (617) 732-5254; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1383–1395, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X513991
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Evaluation of 28 Human Embryonic Stem Cell Lines for Use as Unrelated Donors in Stem Cell Therapy: Implications of HLA and ABO Genotypes

Jeoung Eun Lee,* Myung Seo Kang,† Myoung Hee Park,‡ Sung Han Shim,§ Tae Ki Yoon,§ Hyung Min Chung,*¶ and Dong Ryul Lee*§

*CHA Stem Cell Institute, CHA University, Seoul, Korea
†Department of Laboratory Medicine, College of Medicine, CHA University, Seoul, Korea
‡Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea
§Fertility Center, CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, Korea
¶CHA Bio & Diostech Co., Ltd., Seoul, Korea

For human embryonic stem cells (hESCs) to be used clinically, it is imperative that immune responses evoked by hESCs and their derivates after transplantation should be prevented. Human leukocyte antigens (HLA) and ABO blood group antigens are important histocompatibility factors in graft rejection. HLA matching between recipient and unrelated donors, in particular, is important in improving outcomes in hematopoietic cell transplantation (HCT). We have established and successfully maintained 29 hESC lines and analyzed the HLA and ABO genotypes of these lines. HLA-A, -B, -C and -DR (DRB1) genotyping was performed by polymerase chain reaction (PCR) sequence-based typing and ABO genotyping was carried out by PCR restriction fragment length polymorphism methods. To determine what proportion of the Korean population would be covered by these cell lines in organ transplantation, 27 cell lines with HLA-A, -B, and -DR data were evaluated for HCT (cord blood) donors and 28 cell lines with HLA-DR and ABO data were evaluated for solid organ (kidney) transplantation donors, and then compared the data with those from 6,740 donated cord bloods. When 2 HLA mismatches are allowed for HCT, as currently accepted for cord blood transplantation, it was estimated that about 16% and 25% of the possible recipients can find one or more donor cell lines with ≤2 mismatches at A, B, DRB1 allele level and at A, B antigen/DRB1 allele level, respectively. When HLA-DR antigen level matching and ABO compatibility was considered for solid organ (kidney) transplantation, it was estimated that about 29% and 96% of the possible recipients can find one or more ABO-compatible donor cell lines with 0 and 1 DR mismatches, respectively. We provided the first report on the HLA and ABO genotypes of hESC lines, and estimated the degree of HLA and ABO matching in organ transplantation for the Korean population.

Key words: Human embryonic stem cells (hESCs); Donated cord blood; HLA genotype; ABO genotype; Cell therapy

Address correspondence to Dong Ryul Lee, Ph.D., CHA Stem Cell Institute, CHA University, 606-5 Yeoksam-dong, Gangnam-gu, Seoul, 135-081, Korea. Tel: 82-2-3468-3421; Fax: 82-2-3468-2610; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1397–1412, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X513955
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Cardiac Cell Generation From Encapsulated Embryonic Stem Cells in Static and Scalable Culture Systems

Donghui Jing,* Abhirath Parikh,* and Emmanuel S. Tzanakakis*†

*Department of Chemical and Biological Engineering, State University of New York at Buffalo, Buffalo, NY, USA
†New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, USA

Heart diseases are major causes of morbidity and mortality linked to extensive loss of cardiac cells. Embryonic stem cells (ESCs) give rise to cardiomyocyte-like cells, which may be used in heart cell replacement therapies. Most cardiogenic differentiation protocols involve the culture of ESCs as embryoid bodies (EBs). Stirred-suspension bioreactor cultures of ESC aggregates may be employed for scaling up the production of cardiomyocyte progeny but the wide range of EB sizes and the unknown effects of the hydrodynamic environment on differentiating EBs are some of the major challenges in tightly controlling the differentiation outcome. Here, we explored the cardiogenic potential of mouse ESCs (mESCs) and human ESCs (hESCs) encapsulated in poly-L-lysine (pLL)-coated alginate capsules. Liquefaction of the capsule core led to the formation of single ESC aggregates within each bead and their average size depended on the concentration of seeded ESCs. Encapsulated mESCs were directed along cardiomyogenic lineages in dishes under serum-free conditions with the addition of bone morphogenetic protein 4 (BMP4). Human ESCs in pLL-layered liquid core (LC) alginate beads were also differentiated towards heart cells in serum-containing media. Besides the robust cell proliferation, higher fractions of cells expressing cardiac markers were detected in ESCs encapsulated in LC than in solid beads. Furthermore, we demonstrated for the first time that ESCs encapsulated in pLL-layered LC alginate beads can be coaxed towards heart cells in stirred-suspension bioreactors. Encapsulated ESCs yielded higher fractions of Nkx2.5- and GATA4-positive cells in the bioreactor compared to dish cultures. Differentiated cells formed beating foci that responded to chronotropic agents in an organotypic manner. Our findings warrant further development and implementation of microencapsulation technologies in conjunction with bioreactor cultivation to enable the production of stem cell-derived cardiac cells appropriate for clinical therapies and applications.

Key words: Embryonic stem cells (ESCs); Heart cells; Directed differentiation; Encapsulation; Bioreactor

Address correspondence to Emmanuel S. Tzanakakis, Department of Chemical and Biological Engineering, State University of New York at Buffalo, 907 Furnas Hall, Buffalo, NY 14260, USA. Tel: (716) 645-1201; Fax: (716) 645-3822; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1413–1424, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X514170
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Cell Therapy, a New Standard in Management of Chronic Critical Limb Ischemia and Foot Ulcer

V. Procházka,* J. Gumulec,† F. Jalůvka,‡ D. Šalounová,§ T. Jonszta,* D. Czerný,* J. Krajča,* R. Urbanec,‡ P. Klement,¶ J. Martinek,# and G. L. Klement**

*Radiodiagnostic Institute, University Hospital Ostrava, Ostrava-Poruba, Czech Republic
†Hemato-Oncological Center, University Hospital Ostrava, Ostrava-Poruba, Czech Republic
‡Surgery Clinic and Anaesthesiology Department, University Hospital Ostrava, Ostrava-Poruba, Czech Republic
§Department of Mathematical Methods in Economy, VSˇ B-Technical University Ostrava, Ostrava-Poruba, Czech Republic
¶Henderson Research Center, McMaster University, Hamilton, Ontario, Canada
#Clinical Laboratory, J.G. Mendel Cancer Center, Novy Jicin, Czech Republic
**Children’s Hospital, Dana-Farber Cancer Institute, Harvard Medical University, Boston, MA, USA

Fifty percent of diabetics (7% of general population) suffer from peripheral arterial occlusive disease, which may lead to amputation due to critical limb ischemia (CLI). The aim of our study was to prevent major limb amputation (MLA) in this group of patients using a local application of autologous bone marrow stem cells (ABMSC) concentrate. A total of 96 patients with CLI and foot ulcer (FU) were randomized into groups I and II. Patients in group I (n = 42, 36 males, 6 females, 66.2 ± 10.6 years) underwent local treatment with ABMSC while those in group II (n = 54, control, 42 males, 12 females, 64.1 ± 8.6 years) received standard medical care. The frequency of major limb amputation in groups I and II was 21% and 44% within the 120 days of follow up, respectively (p < 0.05). Only in salvaged limbs of group I both toe pressure and toe brachial index increased (from 22.66 ± 5.32 to 25.63 ± 4.75 mmHg and from 0.14 ± 0.03 to 0.17 ± 0.03, respectively, mean ± SEM). The CD34+ cell counts in bone marrow concentrate (BMC) decreased (correlation, p = 0.024) with age, even though there was no correlation between age and healing. An unexpected finding was made of relative, bone marrow lymphopenia in the initial bone marrow concentrates in patients who failed ABMSC therapy (21% of MLA). This difference was statistically significant (p < 0.040). We conclude ABMSC therapy results in 79% limb salvage in patients suffering from CLI and FU. In the remaining 21% lymphopenia and thrombocytopenia were identified as potential causative factors, suggesting that at least a partial correction with platelet supplementation may be beneficial.

Key words: Critical limb ischemia (CLI); Diabetic foot ulcer; Autologous bone marrow stem cells (ABMSC); Lymphopenia of bone marrow

Address correspondence to Václav Procházka, M.D., Ph.D., M.Sc., Interventional Neuroradiology and Angiology, Radiology Department, Ave 17. Listopadu 1790, University Hospital Ostrava, Ostrava-Poruba, 70852, Czech Republic. Tel: 00420 59737 2172; Fax: 00420 59737 2175; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1425–1437, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X509068
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Beneficial Effects of VEGF Secreted From Stromal Cells in Supporting Endothelial Cell Functions: Therapeutic Implications for Critical Limb Ischemia

Gilda Cobellis,* Ciro Maione,* Chiara Botti,* Antonietta Coppola,* Andrea Silvestroni,* Stefano Lillo,† Vincenzo Schiavone,† Anna Maria Molinari,* and Vincenzo Sica*‡

*Dipartimento di Patologia Generale, Cattedra di Patologia Clinica, Seconda Università degli Studi di Napoli, Napoli, Italy
†Clinica Pineta Grande, Castelvolturno, Caserta, Italy
‡Università Telematica “Guglielmo Marconi”, Roma, Italy

Critical limb ischemia (CLI) is the end stage of peripheral vascular disease (PVD). One third of CLI patients progresses to leg amputation with high associated morbidity and mortality. In no-option patients with end-stage critical limb ischemia, bone marrow cell transplantation has shown promising results, improving leg perfusion to the level of reducing major amputations and allowing limb salvage. We recently reported the successful application of an innovative protocol based on repeated autologous bone marrow cell transplantation, which resulted in an effective and feasible strategy for achieving long-term revascularization in patients with severe CLI. In an effort to understand the clinical benefit provided by stem cells therapy in patients with CLI, we characterized the marrow-derived stromal cells of CLI patients and we provided a correlation between the in vitro features of these cells and the clinical follow up at 12 months. We showed that cells derived from CLI patients had a reduced capacity to proliferate, adhere, and migrate, but that they stimulated proliferation and migration of endothelial cells through the release of VEGF-A, supporting the idea that the paracrine mechanisms underpinned the biological effects of long-term angiogenesis in CLI patients.

Key words: Critical limb ischemia (CLI); Marrow-derived stromal cells; Trophic factors; Vascular endothelial growth factor (VEGF)

Address correspondence to Gilda Cobellis, Ph.D., Dipartimento di Patologia Generale, Cattedra di Patologia Clinica, Seconda Università degli Studi di Napoli, Via L. De Crecchio, 7, 80138 Napoli, Italy. Tel: +390815665681; Fax: +39081292399; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1439–1449, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X514260
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

An Efficient Approach to Isolation and Characterization of Pre- and Postnatal Umbilical Cord Lining Stem Cells for Clinical Applications

R. Gonzalez,* L. Griparic,* M. Umana,* K. Burgee,* V. Vargas,* R. Nasrallah,* F. Silva,* and A. Patel†

*DaVinci Biosciences LLC, Costa Mesa, CA, USA
†Cardiovascular Center, University of Utah, Salt Lake City, UT, USA

There have been various forms of mesenchymal stem cell-like (MSC-like) cells isolated from umbilical cords (UCs). The isolation of umbilical cord lining stem cells (ULSCs) may be of great value for those interested in a possible treatment to several disease/disorders. Unlike umbilical cord blood cells, these cells are unique because they can be expanded to therapeutically relevant numbers and cryopreserved for several different uses. Here we efficiently isolate stem cells from a small segment of pre- and postnatal UCs, and obtain therapeutically relevant amounts of ULSCs within 3 weeks. We demonstrate their growth potential and characterize them using immunocytochemistry, flow cytometry, and RT-PCR. In addition, we differentiate ULSCs into multiple lineages. Pre- and postnatal ULSCs are morphologically similar to mesenchymal stem cells (MSCs) and easily expand to greater than 70 population doublings. They express pluripotent markers Oct4 and nanog at the protein and RNA level. Flow cytometry demonstrates that they express markers indicative of MSCs in addition to high SSEA-4 expression. ULSCs are easily differentiated into osteogenic, adipogenic, chondrogenic, cardiogenic, and neurogenic cells. Pre- and postnatal ULSCs are characteristically similar in respect to their growth, marker expression, and plasticity, demonstrating they are highly conserved throughout development. ULSCs have phenotypic and genotypic properties of MSCs. These studies demonstrate the therapeutic potential of an otherwise discarded tissue. They are a perfect HLA match for the donor and an excellent match for immediate family members; therefore, they may serve as a therapeutic cell source.

Key words: Umbilical cord lining; Mesenchymal stem cells; Differentiation; Multipotent; Cell banking; Clinical applications

Address correspondence to Rafael Gonzalez, Ph.D., Director of Research & Development, DaVinci Biosciences LLC, 1239 Victoria Street, Costa Mesa, CA 92627, USA. Tel: 949-515-2828; Fax: 949-515-2929; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Amit Patel, M.D., M.S., Associate Professor of Surgery, Divison of Cardiothoracic Surgery, Director of Cardiovascular Regenerative Medicine, Director of Clinical Regenerative Medicine, CTF, 30 N 1900E 3C127, Salt Lake City, UT 84132, USA. Tel: 801-581-5311; Fax: 801-585-3936; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1451–1463, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X514198
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Transplantation of Human Wharton’s Jelly-Derived Stem Cells Alleviates Chemically Induced Liver Fibrosis in Rats

Shinn-Zong Lin,*1 Yao-Jen Chang,†1 Jen-Wea Liu,‡ Li-Fu Chang,‡ Li-Yi Sun,‡ Yuan-Sheng Li,‡ Geng-Hong Luo,‡ Chia-Hui Liao,‡ Pin-Hsin Chen,‡ Tse-Min Chen,§ Ru-Ping Lee,¶ Kuo-Liang Yang,# Horng-Jyh Harn,** and Tzyy-Wen Chiou‡

*Center for Neuropsychiatry, China Medical University and Hospital and Beigang Hospital, Taichung and Yun-Lin, Taiwan, ROC
†Department of Surgery, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, ROC
‡Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien, Taiwan, ROC
§Division of Laboratory Medicine, Hualien Armed Forces General Hospital, Hualien, Taiwan, ROC
¶Department of Nursing, Tzu Chi University, Hualien, Taiwan, ROC
#Cord Blood Bank, Tzu Chi Stem Cell Center, Hualien, Taiwan, ROC
**Pathology Department, China Medical University & Hospital, Taichung, Taiwan, ROC

There is currently no effective treatment method available for liver fibrosis. We therefore evaluated the use of Wharton’s jelly stem cells (WJSCs; the major umbilical cord stem cell population) to treat chemically induced liver fibrosis via intraperitoneal injection of thioacetamide. WJSCs were transplanted into liver-damaged rats via the portal vein and the treatment was evaluated by assessing serum biochemistry and histopathology. Transplanted WJSCs were distributed in the fibrotic area and around blood vessels, and hepatic recovery was accelerated. Serum prothrombin time significantly recovered, and serum albumin also improved at 21 days posttransplantation; collagen accumulation also decreased at 14 days. Thus, human WJSCs promoted recovery after chronic liver damage. Using immunohistochemical analyses, we determined that transplanted WJSCs produce albumin, hepatocyte growth factor (HGF), and metalloproteinase (MMP) after transplantation to chemically injured liver, indicating that WJSC may help to decrease liver collagen and thus may be useful for treating liver fibrosis.

Key words: Liver fibrosis; Stem cells; Thioacetamide; Wharton’s jelly; Metalloproteinase

1
These authors provided equal contribution to this study.
Address correspondence to Dr. Tzyy-Wen Chiou, Ph.D., Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University No. 1, Sec. 2, Da Hsueh Rd., Shoufeng, Hualien, Taiwan, Republic of China. Tel: 886-3-8633638; Fax: 886-3-8630262; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it or Dr. Horng-Jyh Harn, Pathology Department, China Medical University & Hospital, 2 Yuh-Der Road, Taichung, Taiwan, 40447, Republic of China. Tel: 886-4-22052121, ext. 2661; Fax: 886-4-22052121, ext. 2566; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1465–1473, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X314161
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

The Assessment of Human Erythroid Output in NOD/SCID Mice Reconstituted With Human Hematopoietic Stem Cells

Jun Hayakawa,* Matthew M. Hsieh,* D. Eric Anderson,† Oswald Phang,* Naoya Uchida,* Kareem Washington,* and John F. Tisdale*

*Molecular and Clinical Hematology Branch, National Institutes of Diabetes and Digestive and Kidney Disorders (NIDDK) and National Heart, Lung, and Blood Institute (NHLBI), National Institutes of Health, Bethesda, MD, USA
†Proteomics and Mass Spectrometry Facility, National Institutes of Diabetes and Digestive and Kidney Disorders (NIDDK), National Institutes of Health, Bethesda, MD, USA

The third-generation NOD/LtSz-scid/IL2Rγnull (NOD/SCID IL2Rγnull) mouse represents a significantly improved xenograft model allowing high levels of human leukocyte engraftment over extended follow up. One remaining limitation of this mouse model, however, is the low level of circulating human erythrocytes. We established a practical ex vivo erythroid culture system of xenograft marrow progenitors to enrich for human erythroid progeny. At various time points after transplant, erythroid cells were easily assayed after 17 days of ex vivo culture of xenograft marrow, with nearly all nucleated cells of human origin and approximately 60% human GPA or CD71 positive. We then transplanted cord blood CD34+ cells marked with a lentiviral vector encoding green fluorescent protein (GFP). Three months later, ex vivo culture of xenograft marrow progenitors showed 41.3% of the cultured erythroid cells were positive for GFP and human CD71, and 56.2% were positive for GFP and human GPA, similar to that of circulating leukocytes at the same time point. Next, G-CSF mobilized peripheral blood CD34+ cells from a sickle cell trait subject were infused in this mouse model to determine if the hemoglobin pattern could be modeled. CD34+ cells from the sickle cell trait subject engrafted equally compared to CD34+ cells from normal subjects, establishing the sickle cell trait phenotype. Lastly, a comparison of adult-derived peripheral blood CD34+ cells and cord blood-derived CD34+ cells xenografted mice was made, and long term follow-up demonstrated a recapitulation of the fetal to adult hemoglobin switch. This approach should prove a useful tool for testing strategies for genetic manipulation of erythroid progeny and the study of hemoglobin switching.

Key words: NOD/LtSz-scid/IL2Rγnull mice; Erythropoiesis; Xenograft mouse model; Sickle cell; HbF; HbS; Hemoglobin switching; Lentiviral vector; GFP

Address correspondence to John F. Tisdale, 9000 Rockville Pike, Building 10, Room 9N112, Bethesda, MD 20892, USA. Tel: 301-402-6497; Fax: 301-480-8975; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1475–1486, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X514314
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Autologous Hematopoietic Stem Cell Transplantation in 48 Patients With End-Stage Chronic Liver Diseases

Hosny Salama,* Abdel-Rahman Zekri,† Mark Zern,‡ Abeer Bahnassy,† Samah Loutfy,† Sameh Shalaby,† Cheryl Vigen,§ Wendy Burke,§ Mohamed Mostafa,¶ Eman Medhat,* Omar Alfi,# and Elizabeth Huttinger**

*Hepatology Department, Cairo University Hospital, Cairo, Egypt
†National Cancer Institute, Cairo, Egypt
‡University of California Davis Medical Center, Sacramento, CA, USA
§University of Southern California/Keck School of Medicine, Los Angeles, CA, USA
¶Radiology Department, Cairo University Hospital, Cairo, Egypt
#Alfi Stem Cell Research and Education Institute, Pasadena, CA, USA
**International Study Group for Stem Cell Therapy, Pasadena, CA, USA

The only presently viable treatment for end-stage liver disease is whole organ transplantation. However, there are insufficient livers available. The aim of the present study is to provide autologous bone marrow-derived stem cells as a potential therapeutic for patients with end-stage cirrhosis. This is a retrospective chart review of autologous stem cell treatment in 48 patients, 36 with chronic end-stage hepatitis C-induced liver disease and 12 with end-stage autoimmune liver disease. For all patients, granulocyte colony-stimulating factor was administered to mobilize their hematopoietic stem cells. Following leukapheresis, CD34+ stem cells were isolated, amplified, and partially differentiated in culture, then reinjected into each subject via their hepatic artery or portal vein. Treatment was generally well tolerated with the expected moderate but transient bone pain from G-CSF in less than half of the patients. Three patients had serious treatment-related complications, and only 20.8% of these end-stage liver disease patients died during 12 months of follow up. For all patients there was a statistically significant decrease in ascites. There was clinical and biochemical improvement in a large percentage of patients who received the transplantation. In the viral group, there were marked changes in albumin (p = 0.0003), bilirubin (p = 0.04), INR (p = 0.0003), and ALT levels (p = 0.02). In the autoimmune group, values also improved significantly for albumin (p = 0.001), bilirubin (p = 0.002), INR (p = 0.005), and ALT levels (p = 0.003). These results suggest that autologous CD34+ stem cell transplantation may be safely administered and appears to offer some therapeutic benefit to patients with both viral and autoimmune-induced end-stage liver disease.

Key words: Cell transplantation; Cirrhosis; Hematopoietic stem cells

Address correspondence to Mark A. Zern, M.D., UC Davis Medical Center, 4635 Second Ave Ste. 1001, Sacramento, CA 95817, USA. Tel: (916) 734-8063; Fax: (916) 734-8097; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1487–1492, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X514620
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Brief Communication

N
-Acetylcysteine Improves the Viability of Human Hepatocytes Isolated From Severely Steatotic Donor Liver Tissue

Filippos G. Sagias,*1 Ragai R. Mitry,*1 Robin D. Hughes,* Sharon C. Lehec,* Ameet G. Patel,† Mohamed Rela,* Giorgina Mieli-Vergani,* Nigel D. Heaton,* and Anil Dhawan*

*Institute of Liver Studies, King’s College London School of Medicine at King’s College Hospital, London, UK
†Hepatopancreatobiliary Surgery Service, King’s College Hospital, London, UK

Hepatocyte transplantation is dependent on the availability of good quality human hepatocytes isolated from donor liver tissue. Hepatocytes obtained from livers rejected for transplantation on the grounds of steatosis are often of low viability and not suitable for clinical use. The aim of this study was to evaluate the effects of the antioxidant N-acetylcysteine (NAC) on the function of hepatocytes isolated from steatotic donor livers. Human hepatocytes were isolated from 10 severely steatotic (>60%) donor livers rejected for transplantation. The left lateral segment of the donor liver was dissected into two equal size pieces and randomized to NAC or control. NAC (5 mM) was added to the first perfusion buffer of the standard collagenase digestion technique. Cells from tissues perfused with NAC had a significantly higher mean viability (81.1 ± 1.7% vs. 66.0 ± 4.7%; p = 0.003) and cell attachment (1.08 ± 0.26 vs. 0.67 ± 0.18 OD units; p = 0.012). Addition of NAC during isolation of human hepatocytes from steatotic donor liver tissue significantly improved the outcome of cell isolation. Further studies are needed to investigate the mechanism(s) of this effect. Incorporation of NAC in the hepatocyte isolation protocol could increase the availability of hepatocytes for transplantation.

Key words: Steatosis; Hepatocyte transplantation; N-Acetylcysteine; Donor liver

1
These authors provided equal contribution to this work.
Address correspondence to Professor Anil Dhawan, Paediatric Liver Centre, King’s College Hospital, Denmark Hill, London SE5 9RS, UK. Tel: +44-20-3299-3214; Fax: +44-20-3299-3564; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Transplantation
, Vol. 19, pp. 1493–1509, 2010
0963-6897/10 $90.00 + .00
DOI: 10.3727/096368910X540612
E-ISSN 1555-3892
Copyright © 2010 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Editorial Commentary

Stem Cell Research in
Cell Transplantation: Sources, Geopolitical Influence, and Transplantation

David J. Eve, Randolph W. Fillmore, Cesar V. Borlongan, and Paul R. Sanberg

Center of Excellence for Aging & Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida College of Medicine, Tampa, FL, USA

If the rapidly progressing field of stem cell research reaches its full potential, successful treatments and enhanced understanding of many diseases are the likely results. However, the full potential of stem cell science will only be reached if all possible avenues can be explored and on a worldwide scale. Until 2009, the US had a highly restrictive policy on obtaining cells from human embryos and fetal tissue, a policy that pushed research toward the use of adult-derived cells. Currently, US policy is still in flux, and retrospective analysis does show the US lagging behind the rest of the world in the proportional increase in embryonic/fetal stem cell research. The majority of US studies being on either a limited number of cell lines, or on cells derived elsewhere (or funded by other sources than Federal) rather than on freshly isolated embryonic or fetal material. Neural, mesenchymal, and the mixed stem cell mononuclear fraction are the most commonly investigated types, which can generally be classified as adult-derived stem cells, although roughly half of the neural stem cells are fetal derived. Other types, such as embryonic and fat-derived stem cells, are increasing in their prominence, suggesting that new types of stem cells are still being pursued. Sixty percent of the reported stem cell studies involved transplantation, of which over three quarters were allogeneic transplants. A high proportion of the cardiovascular systems articles were on allogeneic transplants in a number of different species, including several autologous studies. A number of pharmaceutical grade stem cell products have also recently been tested and reported on. Stem cell research shows considerable promise for the treatment of a number of disorders, some of which have entered clinical trials; over the next few years it will be interesting to see how these treatments progress in the clinic.

Key words: Stem cells; Cell transplantation; Regenerative medicine; Allogeneic; Autologous

Address correspondence to Dr. David Eve, Center of Excellence for Aging and Brain Repair, Department of Neurosurgery and Brain Repair, University of South Florida, 12901 Bruce B Downs Blvd., MDC-78, Tampa, FL 33612, USA. Tel: 813-974-6169; Fax: 813-974-3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it