ognizant Communication Corporation

CELL TRANSPLANTATION
The Regenerative Medicine Journal

ABSTRACTS
VOLUME 16, NUMBER 5, 2007

Cell Transplantation, Vol. 16, pp. 449-459, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Nitric Oxide-Containing Neurons in Long-Term Grafts in a Rat Model of Parkinson'S Disease

B. Rajakumar,1 B. A. Flumerfelt,1 A. W. Hrycyshyn,1 and N. Rajakumar1,2

1Department of Anatomy & Cell Biology, University of Western Ontario, London, Ontario, Canada
2Department of Psychiatry, University of Western Ontario, London, Ontario, Canada

The role that nitric oxide may play in modulating graft function in long-term fetal ventral mesencephalic grafts in an animal model of Parkinson's disease was investigated. Mature grafts harvested from the entire fetal ventral mesencephalon possessed a large number of neuronal nitric oxide synthase (nNOS)\NADPH-diaphorase-containing neurons throughout the graft intermingled with dopaminergic neurons. The morphological and neurochemical characteristics of these NADPH-diaphorase neurons resembled those in centers adjacent to the substantia nigra of adult brain but not that of the striatum. Pretreatment with the nNOS blocker, 7-nitroindazole, resulted in contralateral rotations following methamphetamine challenge in long-term grafted animals that previously showed normalized rotational behavior. In contrast, mature grafts derived from fetal ventral mesencephalon without the midline areas possessed only a few nNOS-containing neurons within the grafts, and a similar methamphetamine challenge following 7-nitroindazole pretreatment in long-term grafted rats that previously showed normalized rotational behavior resulted in random movements. Our results indicate that nitric oxide-containing neurons inadvertently included during grafting may affect graft function, and excluding the midline areas of the ventral mesencephalon during tissue harvesting may minimize this effect.

Key words: Neural tissue transplantation; Ventral mesencephalon; Dopamine; Striatum; NADPH-diaphorase; Nitric oxide synthase

Address correspondence to B. A. Flumerfelt, Ph.D., Department of Anatomy & Cell Biology, University of Western Ontario, London, ON., Canada N6A 5C1. E-mail: bflumer@uwo.ca




Cell Transplantation, Vol. 16, pp. 461-474, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

The Effect of Truncated Human a-Synuclein (1-120) on Dopaminergic Cells in a Transgenic Mouse Model of Parkinson's Disease

A. W. Michell, G. K. Tofaris, H. Gossage, P. Tyers, M. G. Spillantini, and R. A. Barker

Department of Clinical Neuroscience, University of Cambridge and Cambridge Centre for Brain Repair, Cambridge, CB2 2PY, UK

a-Synuclein is thought to play an important role in the pathology of Parkinson's disease (PD). Truncated forms of this protein can be found in PD brain extracts, and these species aggregate faster and are more susceptible to oxidative stress than the full-length protein. We investigated the effect of truncated a-synuclein on dopaminergic cells using a transgenic mouse expressing a-synuclein (1-120) driven by the rat tyrosine hydroxylase promoter on a mouse a-synuclein null background. We found a selective reduction in the yield of dopaminergic cells from transgenic embryonic ventral mesencephalic cell cultures. However, in vivo the substantia nigra\ventral tegmentum dopaminergic cell counts were not reduced in transgenics, although these mice are known to have reduced striatal dopamine. When transplanted to the striatum in the unilateral 6-hydroxydopamine-lesioned mouse model of PD, dopaminergic cells derived from transgenic embryonic ventral mesencephala were significantly smaller at 6 weeks, and showed a trend towards being less effective at ameliorating rotational asymmetry than those from control a-synuclein null mice. These results suggest that a-synuclein (1-120) renders dopaminergic cells more susceptible to stress, which may have important implications as to how this truncated protein might contribute to dopaminergic cell death in sporadic PD.

Key words: a-Synuclein; Transplant; Parkinson's disease; Dopamine

Address correspondence to A. W. Michell, Cambridge Centre for Brain Repair and University of Cambridge Department of Clinical Neuroscience, Forvie Site, Robinson Way, Cambridge, CB2 2PY, UK. Tel: +44 1223 331160; Fax: +44 1223 331174; E-mail: awmichell@gmail.com




Cell Transplantation, Vol. 16, pp. 475-481, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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Combined Treatment of Neurotrophin-3 Gene and Neural Stem Cells Is Propitious to Functional Recovery After Spinal Cord Injury

Luyi Zhang,1 Shuting Gu,1,2 Cuiping Zhao,1 and Tieqiao Wen1,2

1Laboratory of Molecular Neural Biology, School of Life Sciences, Shanghai University, Shanghai 200444, China
2Institute of Systems Biology, Shanghai University, Shanghai 200444, China

We present an insight of the effects of combination therapy with neurotrophin-3 and neural stem cell on functional recovery after spinal cord injury (SCI). Total RNA was extracted from neural stem cell line C17.2 and reversed transcribed into cDNA. Neurotrophin-3 (NT-3) gene was amplified by PCR and subcloned into plasmid to construct an expression vector pNT-3. A positive clone containing pNT-3, named SHN2, was obtained and used for transplantation. Thirty adult mice received mechanical injury at the T8 vertebra level. Cell survival, NT-3 gene expression, and functional recovery were observed through X-Gal staining, RT-PCR, and open field locomotion, respectively. The results show that NT-3 gene comprising 777 bp nucleotides was cloned and a more than twofold expression was detected when transfected into neural stem cell line C17.2. Quantitative analysis of cellular density revealed a significant increase in SHN2 compared to the control cells (p < 0.01). Thirty days after transplantation, SHN2 showed significant increase near the lesion site. Furthermore, the functional recovery indicated an active effect by detecting Basso-Beattie-Bresnahan (BBB) locomotor rating scale (p < 0.01). In conclusion, combined treatment of neural stem cells and NT-3 gene can facilitate functional recovery. It offers an effective approach to treat SCI.

Key words: Neural stem cells; Neurotrophis-3; Spinal cord injury; Transplantation; Functional recovery

Address correspondence to Tieqiao Wen, School of Life Sciences, Shanghai University, 99 Shangda Road, Shanghai, 200444, China. Tel: +86-21-66134717; Fax: +86-21-66132512; E-mail: tqwen@staff.shu.edu.cn




Cell Transplantation, Vol. 16, pp. 483-491, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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An Immortalized Rat Ventral Mesencephalic Cell Line, RTC4, Is Protective in a Rodent Model of Stroke

B. K. Harvey,1 G. J. Chen,1 C. J. Schoen,2 C. T. Lee,2 D. B. Howard,1 O. Dillon-Carter,2 M. Coggiano,2 W. J. Freed,2 Y. Wang,1 B. J. Hoffer,2 and J. F. Sanchez2

1Molecular Neuropsychiatry Research Branch, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), Baltimore, MD, USA
2Cellular Neurobiology Research Branch, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), Baltimore, MD, USA

One therapeutic approach to stroke is the transplantation of cells capable of trophic support, reinnervation, and\or regeneration. Previously, we have described the use of novel truncated isoforms of SV40 large T antigen to generate unique cell lines from several primary rodent tissue types. Here we describe the generation of two cell lines, RTC3 and RTC4, derived from primary mesencephalic tissue using a fragment of mutant T antigen, T155c (cDNA) expressed from the RSV promoter. Both lines expressed the glial markers vimentin and S100b, but not the neuronal markers NeuN, MAP2, or b-III-tubulin. A screen for secreted trophic factors revealed substantially elevated levels of platelet-derived growth factor (PDGF) in RTC4, but not RTC3 cells. When transplanted into rat cortex, RTC4 cells survived for at least 22 days and expressed PDGF. Because PDGF has been reported to reduce ischemic injury, we examined the protective functions of RTC4 cells in an animal model of stroke. RTC4 or RTC3 cells, or vehicle, were injected into rat cortex 15-20 min prior to a 60-min middle cerebral artery ligation. Forty-eight hours later, animals were sacrificed and the stroke volume was assessed by triphenyl-tetrazolium chloride (TTC) staining. Compared to vehicle or RTC3 cells, transplanted RTC4 cells significantly reduced stroke volume. Overall, we generated a cell line with glial properties that produces PDGF and reduces ischemic injury in a rat model of stroke.

Key words: Platelet-derived growth factor (PDGF); Stroke; Ischemia; Transplantation; Ventral mesencephalon

Address correspondence to William J. Freed, Ph.D., NIDA Intramural Research Program, 333 Cassell Drive, Triad Bldg, Room 3501, Baltimore, MD, 21224, USA. Tel.: 410-550-6565, ext. 131; Fax: 410-550-1621; E-mail: wfreed@mail.nih.gov




Cell Transplantation, Vol. 16, pp. 493-503, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Chondroitinase ABC Treatment Enhances Synaptogenesis Between Transplant and Host Neurons in Model of Retinal Degeneration

Takuya Suzuki,1 Masayuki Akimoto,2 Hiroo Imai,3 Yoshiki Ueda,4 Michiko Mandai,2 Nagahisa Yoshimura,1 Anand Swaroop,5,6 and Masayo Takahashi2

1Department of Opthalmology and Visual Sciences, Graduate School of Medicine, Kyoto University, Kyoto, Japan
2Department of Experimental Therapeutics, Translational Research Center, Kyoto University Hospital, Kyoto, Japan
3Department of Biophysics, Graduate School of Science, Kyoto University, Kyoto, Japan
4Department of Ophthalmology, Nagahama City Hospital, Nagahama, Shiga, Japan
5Department of Ophthalmology & Visual Sciences, W.K. Kellogg Eye Center, University of Michigan, Ann Arbor, MI, USA
6Department of Human Genetics, W.K. Kellogg Eye Center, University of Michigan, Ann Arbor, MI, USA

Although recent studies revealed chondroitinase ABC (ChABC), an enzyme that degrades chondroitin sulfate proteoglycans, promotes CNS regeneration in vivo, the usefulness of its application for transplantation is not clear. We investigated if treatment with ChABC can promote synapse formation between graft and host neurons following retinal transplantation. Dissociated retinal cells were prepared from neonatal Nrl-GFP transgenic mice in which rod photoreceptors and their progenitor cells are labeled with GFP. Each cell suspension with or without ChABC (Nrl\ChABC group and Nrl group, respectively) was injected subretinally into the eyes of mice following chemically induced photoreceptor degeneration. The survival and functional integration of the transplanted photoreceptors were examined by histologically and electrophysiologically. Up to 4 weeks after transplantation, almost all the grafted GFP+ photoreceptor cells were widely distributed at the outer margin of the host retina where the photoreceptor layer was located originally. In the Nrl\ChABC group, 33.6% of the GFP+ photoreceptors elaborated neurites horizontally or vertically, and 4.6% elaborated neurites toward the retina. These neurites extended over the glial seal at the graft-host interface, and established synaptic contacts with neurons in the host retina as determined by confocal microscopy and three-dimensional analysis. Although 30.7% cells (p = 0.68) elaborated neurites in the Nrl group, only 1.2% cells (<i>p<i> < 0.05) projected neurites towards the host tissue and synaptic contacts were rare. Our results illustrate the potential utility of ChABC for enhancing synaptogenesis between transplanted neurons and host retina.

Key words: Retinal degeneration; Retinal transplantation; Chondoritinase ABC; Synaptogenesis; Remodeling; Glial scar

Address correspondence to Masayuki Akimoto, Department of Experimental Therapeutics, Translational Research Center, Kyoto University Hospital, Kyoto, 606-8507, Japan. Tel: +81-75-751-4717; Fax: +81-75-751-4731; E-mail: masayuki@akimoto3.com




Cell Transplantation, Vol. 16, pp. 505-516, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Time Course and Quantification of Pancreatic Islet Revasculariztion Following Intraportal Transplantation

Gareth L. Jones,1 Maciej T. Juszczak,2 Stephen J. Hughes,3 Paul Kooner,3 Stephen H. Powis,1 and Martin Press 2

1Centre for Nephrology, Royal Free Campus, Royal Free and University College Medical School, London, NW3 2PF, UK
2Department of Endocrinology, Royal Free Campus, Royal Free and University College Medical School, London, NW3 2PF, UK
3Department of Hepatology, Royal Free Campus, Royal Free and University College Medical School, London, NW3 2PF, UK

A large proportion of islets are lost after transplantation partly due to a lack of functional vasculature. Islets revascularize from host tissue but the process takes up to 2 weeks and has been suggested to result in reduced vascular density in engrafted islets. We describe a method for observing and quantifying the revascularization of intraportally transplanted islets that includes number, density, and branching of islet capillaries. Syngeneic islets were transplanted selectively into the two right posterior lobes of the liver of adult Lewis rats. Sections of the livers were dual stained for insulin and Bandeiraea simplicifolia and analyzed for islet morphology, area, and vascular density from day 0 to day 14 posttransplant and compared to native islets. Vascular density was 1431 ± 75.7 vessels\mm2 in native islets and fell to 325.3 ± 30.8 vessels\mm2 (p < 0.001) by day 1 posttransplant and subsequently increased until day 14 when it was significantly higher than in native islets (2612.5 ± 107.8 vessels\mm2, p < 0.001). The percentage of islet area occupied by vascular space was 9.1 ± 0.9% in native islets. After falling to 2.3 ± 0.3% (p < 0.001) 1 day posttransplant this rose to supranormal levels (21.5 ± 0.8%, p < 0.001) by day 14. The index of capillary branching was 0.771 ± 0.017 in native islets and fell to 0.465 ± 0.02 (p = 0.001) by day 3 but returned to native values by day 7 posttransplantation (0.726 ± 0.03). This technique provides a robust method for tracking and quantifying the revascularization of intraportally transplanted islets, which should enable the comparison of different strategies aimed at accelerating islet revascularization.

Key words: Islet transplantation; Vascular density; Endothelium; Quantification

Address correspondence to Dr. Martin Press, Department of Endocrinology, Royal Free and University College Medical School, Rowland Hill Street, London, NW3 2PF, UK. Tel: +44 20 7830 2475; Fax: +44 20 7830 2475; E-mail: martin.press@royalfree.nhs.uk and copy to gl.jones@medsch.ucl.ac.uk




Cell Transplantation, Vol. 16, pp. 517-525, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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Role of Blood Glucose in Cytokine Gene Expression in Early Syngeneic Islet Transplantation

Marta Montolio, Noèlia Téllez, Joan Soler, and Eduard Montanya

Laboratory of Diabetes and Experimental Endocrinology, Department of Clinical Sciences, University of Barcelona, and Endocrine Unit, Hospital Universitari de Bellvitge, Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), Barcelona, Spain

In islet transplantation, local production of cytokines at the grafted site may contribute to the initial nonspecific inflammation response. We have determined whether the metabolic condition of the recipient modulates the cytokine expression in islet grafts in the initial days after transplantation. Normoglycemic and hyperglycemic streptozotocin-diabetic Lewis rats were transplanted with 500 syngeneic islets, an insufficient beta cell mass to restore normoglycemia in hyperglycemic recipients. The expression of IL-1b, TNF-a, IFN-g, IL-6, IL-10, and IL-4 genes was determined by real-time PCR in freshly isolated islets, in 24-h cultured islets and in islet grafts on days 1, 3, and 7 after transplantation. IL-1b mRNA was strongly and similarly increased in normoglycemic and hyperglycemic groups on days 1, 3, and 7 after transplantation compared with freshly isolated and cultured islets. TNF-a mRNA was also strongly increased on day 1, and it remained increased on days 3 and 7. IL-6 and IL-10 were not detected in freshly isolated islets, but their expression was clearly enhanced in 24-h cultured islets and islet grafts. IL-6 was further increased in hyperglycemic grafts. IL-10 expression was increased in both normoglycemic and hyperglycemic grafts on day 1 after transplantation, and remained increased in hyperglycemic grafts compared to 24-h cultured islets. IFN-g mRNA was barely detected in a few grafts, and IL-4 mRNA was never detected. Thus, the inflammatory response in islet grafts was maximal on day 1 after transplantation, it was sustained, although at lower levels, on days 3 and 7, and it was partly enhanced by hyperglycemia.

Key words: Islet transplantation; Cytokine; Real-time PCR; Hyperglycemia

Address correspondence to Eduard Montanya, Endocrine Unit (13-2), Hospital Universitari Bellvitge, Feixa Llarga, s\n, 08907 L'Hospitalet de Llobregat, Barcelona. Spain. Tel: +34-934037265; Fax: +34-934035804; E-mail: montanya@ub.edu




Cell Transplantation, Vol. 16, pp. 527-537, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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Cell Loss During Pseudoislet Formation Hampers Profound Improvements in Islet Lentiviral Transduction Efficacy for Transplantation Purposes

H. Callewaert,1 C. Gysemans,1 A. K. Cardozo,2 M. Elsner,3 M. Tiedge,3,4 D. L. Eizirik,2 and C. Mathieu1

1Laboratory of Experimental Medicine and Endocrinology (LEGENDO), UZ Gasthuisberg O&N, Katholieke Universiteit Leuven, Leuven, Belgium
2Laboratory of Experimental Medicine, Université Libre de Bruxelles, Brussels, Belgium
3Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany
4Institute of Medical Biochemistry and Molecular Biology, University of Rostock, Rostock, Germany

Islet transplantation is a promising treatment in type 1 diabetes, but the need for chronic immunosuppression is a major hurdle to broad applicability. Ex vivo introduction of agents by lentiviral vectors--improving &beta;-cell resistance against immune attack--is an attractive path to pursue. The aim of this study was to investigate whether dissociation of islets to single cells prior to viral infection and reaggregation before transplantation would improve viral transduction efficacy without cytotoxicity. This procedure improved transduction efficacy with a LV-pWPT-CMV-EGFP construct from 11.2 ± 4.1% at MOI 50 in whole islets to 80.0 ± 2.8% at MOI 5. Viability (as measured by Hoechst\PI) and functionality (as measured by glucose challenge) remained high. After transplantation, the transfected pseudoislet aggregates remained EGFP positive for more than 90 days and the expression of EGFP colocalized primarily with the insulin-positive &beta;-cells. No increased vulnerability to immune attack was observed in vitro or in vivo. These data demonstrate that dispersion of islets prior to lentiviral transfection and reaggregation prior to transplantation is a highly efficient way to introduce genes of interest into islets for transplantation purposes in vitro and in vivo, but the amount of b-cells needed for normalization of glycemia was more than eightfold higher when using dispersed cell aggregates versus unmanipulated islets. The high price to pay to reach stable and strong transgene expression in islet cells is certainly an important cell loss.

Key words: Type 1 diabetes; Pancreatic b-cells; Pseudoislet aggregates; Gene therapy; Lentiviral vector; Islet transplantation

Address correspondence to Prof. Dr. Chantal Mathieu, LEGENDO, UZ Gasthuisberg O&N1, Herestraat 49, bus 902, B-3000 Leuven, Belgium. Tel: 00-32-16-34.59.70; Fax: 00-32-16-34.59.34; E-mail: chantal.mathieu@med.kuleuven.be




Cell Transplantation, Vol. 16, pp. 539-545, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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Improved Quantity and In Vivo Function of Islets Isolated by Reduced Pressure-Controlled Injection of Collagenase in a Rat Model

Shiri Li,1 Tetsuya Sakai,1,2 Yasuyuki Suzuki,1 Tadahiro Goto,1 Tomohiro Tanaka,1 Takuro Yoshikawa,1 Keitaro Kakinoki,1 Yasuki Tanioka,1,2 Ippei Matsumoto,1 Yasuhiro Fujino,1 and Yoshikazu Kuroda1,2

1Department of Gastroenterological Surgery, Kobe University Graduate School of Medicine, Kobe, Japan
221st Century COE Program, Kobe University Graduate School of Medicine, Kobe, Japan

In islet transplantation, insufficient yield is a major obstacle to one-donor\one-recipient transplant. Collagenase, which is injected via a pancreatic duct to separate islets from acini, can so easily distribute into the islet core that it may result in disruption of islets. The purpose of this study was to evaluate the superiority of reduced pressure-controlled collagenase injection (RPCI) at 80 mmHg on islet isolation to injection at 180 mmHg by examining in vivo transplant experiments besides the yield and the glucose stimulation test in a rat model. Lewis rat pancreases were distended with collagenase solution at 80 mmHg pressure as the RPCI group (group 1) and at 180 mmHg (group 2), followed by isolation. The yield in group 1 (1100 ± 160 islets with 2750 ± 530 IEQ) was significantly higher than that in group 2 (900 ± 130 islets with 1570 ± 350 IEQ, p < 0.01) due to the significant difference of the number of islets sized > 150 mm in diameter, although the purity was not significantly different between the two groups. Stimulation indices in the glucose stimulation tests were 2.88 ± 1.12 in group 1 and 1.93 ± 0.62 in group 2 (p < 0.05). The cure rate by transplantation of 100 islets to diabetic nude mice in group 1 (8\10) was significantly higher than that in group 2 (3\10, p < 0.05). In a syngenic transplant model of 90% of islets isolated from one donor, the cure rates were 100% and 67% in groups 1 and 2, respectively (NS). The area under the curve on the graph of IPGTT on postoperative day 28 in group 1 was significantly smaller than that in group 2 (p < 0.05). In conclusion, our data show that RPCI at 80 mmHg could contribute to consistently high islet yield and in vivo function in a rat model. It was suggested that the current human protocol should be reviewed from this viewpoint.

Key words: Islet isolation; Islet transplantation; Reduced pressure-controlled injection of collagenase

Address correspondence to Tetsuya Sakai, M.D., Ph.D., Department of Gastroenterological Surgery, Kobe University Graduate School of Medicine, 7-5-2 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan. Tel: +81-78-382-5925; Fax: +81-78-382-5939; E-mail: sakai005@nifty.com




Cell Transplantation, Vol. 16, pp. 547-553, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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Intrasplenic Hepatocyte Transplantation Prolonged the Survival in Nagase Analbuminemic Rats With Liver Failure Induced by Common Bile Duct Ligation

Masahiro Ito,1 Hideo Nagata,1 Toshiyuki Yamamoto,1 Daisuke Yoshihara,1 Ira J. Fox,2 and Shuichi Miyakawa1

1Department of Surgery, Fujita-Health University, Toyoake, Aichi, Japan
2Department of Surgery, University of Nebraska Medical Center, Omaha, NE, USA

It has already been established that hepatocyte transplantation (HTx) in animal models, such as both chemically and surgically induced acute liver failure, liver-based metabolic disease, and cirrhosis, resulted in significant improvement of liver function and survival. However, the efficacy of hepatocyte transplantation in secondary cholestatic liver disease is not well known. In this study, we transplanted hepatocytes into the spleen of Nagase analbuminemic rats (NARs) with common bile duct ligation (CBDL) to evaluate the function of transplanted hepatocytes by both of serum albumin levels and total bilirubin levels. CBDL was carried out on NARs to induce liver failure. Lewis rat hepatocytes were transplanted in NARs 7 days after CBDL. Animals, in groups of four, underwent the following interventions: group 1--intrasplenic transplantation of 30 x 106 primary Lewis rat hepatocytes in NARs with CBDL (n = 4), group 2--intrasplenic injection of 0.5 ml DMEM in NARs with CBDL (n = 4); group 3--CBDL only (n = 4); group 4--intrasplenic transplantation of 30 x 106 primary Lewis rat hepatocytes in NARs (n = 4). Both bilirubin levels and albumin levels in NARs with CBDL were significantly improved post-HTx. Animals receiving hepatocyte transplantation survived longer than animals in nontransplant control groups. This study indicates that hepatocytes can be transplanted to temporarily provide life-supporting liver-specific metabolic function and prolong the survival in recipient rats with liver failure induced by CBDL.

Key words: Hepatocyte transplantation; Jaundice; Nagase analbuminemic rats (NARs)

Address correspondence to Masahiro Ito, M.D., Department of Surgery, Fujita Health University, Toyoake City, Aichi, Japan. Tel: (+81) 562-93-9246; Fax: (+81) 562-93-0109; E-mail: masito911@yahoo.co.jp




Cell Transplantation, Vol. 16, pp. 555-562, 2007
0963-6897/07 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2007 Cognizant Comm. Corp.
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Comparison of Mesenchymal Stem Cells From Different Tissues to Suppress T-Cell Activation

Kirsten A. Keyser,1 Karen E. Beagles,1 and Hans-Peter Kiem1,2

1Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
2Departments of Medicine and Pathology, University of Washington School of Medicine, Seattle, WA 98195, USA

Graft-versus-host disease (GvHD) and graft rejection have remained significant complications of allogeneic stem cell transplantation. Mesenchymal stem cells (MSCs) from the bone marrow have been shown to suppress T-cell activation in vitro and in vivo, and may be used to reduce GvHD in the recipient or to facilitate engraftment across MHC barriers. MSCs can be derived from a variety of tissues. Thus, we asked whether MSCs from different tissues might have differential effects on T-cell responses. We were particularly interested in MSCs derived from adipose tissue because of its abundance and accessibility. We investigated and compared the immunosuppressive potential of murine MSCs derived from muscle tissue, adipose tissue, omentum, and bone. Cells from the different tissues were enriched for MSCs and cultured for 2-3 weeks to deplete hematopoietic cells. Mixed lymphocyte reactions (MLRs) including MSCs were performed using concanavalin A or allogeneic T cells as inducers of T-cell activation. MSCs from all tissues differentiated into multiple lineages. Mitogen-induced T-cell activation, as well as allogeneic T-cell responses, was reduced in MLRs mediated by the addition of MSCs. Reduction of T-cell activation was most pronounced for muscle tissue in the mitogen-induced MLR and fat tissue during the allogeneic MLR. These data demonstrate that MSCs from multiple tissues efficiently reduce T-cell activation. The results suggest that MSCs from adipose tissue can serve as an alternative source for MSCs to bone or bone marrow for the modulation of GvHD after allogeneic stem cell transplantation or to enhance engraftment across MHC barriers.

Key words: Mesenchymal stem cells (MSCs); T-cell proliferation; Graft-versus-host disease (GvHD); Immunosuppression; Adipose tissue

Address correspondence to Hans-Peter Kiem, M.D., Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N, Mailstop D1-100, P.O. Box 19024, Seattle, WA 98109-1024, USA. Tel: (206) 667-4425; Fax: (206) 667-6124; E-mail: hkiem@fhcrc.org