ognizant Communication Corporation

The Regenerative Medicine Journal

VOLUME 17, NUMBER 3, 2008

Cell Transplantation, Vol. 16, pp. 241-243, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Tissue Engineering and Biomaterials in Regenerative Medicine

Katherine Nolan,1,4 Yoann Millet,1,4 Camillo Ricordi,1,2,3,5,6 and Cherie L. Stabler1,2,3

1Diabetes Research Institute, University of Miami, Miami, FL, USA
2Department of Surgery, University of Miami, Miami, FL, USA
3Department of Biomedical Engineering, University of Miami, Miami, FL, USA
4Department of Biology, Cornell University, Ithaca, NY, USA
5Karolinska Institute, Stockholm, Sweden
6Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC, USA

The field of regenerative medicine offers the potential to significantly impact a wide spectrum of healthcare issues, from diabetes to cardiovascular disease. In particular, the design of tailored biomaterials, which possess properties desired for their particular application, and the development of superior implant environments, which seek to meet the nutritional needs of the tissue, have yielded promising tissue engineering prototypes. In this commentary, we examine the novel approaches researchers have made in customized biomaterials and promoting angiogenesis that have led to significant advancements in recent years.

Key words: Regenerative medicine; Tissue engineering; Biomaterials; Angiogenesis

Address correspondence to Cherie Stabler, Ph.D., Assistant Professor, Department of Biomedical Engineering, Director, Tissue Engineering Program, Diabetes Research Institute, Miller School of Medicine, University of Miami, 1450 NW 10 Avenue, R-3016, Miami, FL 33136, USA. Tel: (305) 243-9768; Fax: 305-243-4404; E-mail: CStabler@med.miami.edu

Cell Transplantation, Vol. 16, pp. 245-254, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Bioengineering Neural Stem/Progenitor Cell-Coated Tubes for Spinal Cord Injury Repair

Tasneem Zahir,1,3 Hiroshi Nomura,2 Xiao Dong Guo,2 Howard Kim,1 Charles Tator,2,4 Cindi Morshead,1,4 and Molly Shoichet1,3

1Terrence Donnelly Centre for Cellular & Biomolecular Research, University of Toronto, Toronto, Canada
2Toronto Western Research Institute, Toronto Western Hospital, Toronto, Canada
3Department of Chemical Engineering & Applied Chemistry, University of Toronto, Toronto, Canada
4Department of Surgery, University of Toronto, Toronto, Canada

The aim of this study was to understand the survival and differentiation of neural stem/progenitor cells (NSPCs) cultured on chitosan matrices in vivo in a complete transection model of spinal cord injury. NSPCs were isolated from the subependyma of lateral ventricles of adult GFP transgenic rat forebrains. The GFP-positive neurospheres were seeded onto the inner lumen of chitosan tubes to generate multicellular sheets ex vivo. These bioengineered neurosphere tubes were implanted into a completely transected spinal cord and assessed after 5 weeks for survival and differentiation. The in vivo study showed excellent survival of NSPCs, as well as differentiation into astrocytes and oligodendrocytes. Importantly, host neurons were identified in the tissue bridge that formed within the chitosan tubes and bridged the transected cord stumps. The excellent in vivo survival of the NSPCs coupled with their differentiation and maintenance of host neurons in the regenerated tissue bridge demonstrates the promise of the chitosan tubes for stem cell delivery and tissue regeneration.

Key words: Neural stem/progenitor cells; Chitosan; Tissue regeneration; Differentiation; Spinal cord injury

Address correspondence to Molly Shoichet, Terrence Donnelly Centre for Cellular & Biomolecular Research, University of Toronto, 160 College Street, Room 514, Toronto, ON, Canada M5S 3E1. Tel: 416 978 1460; Fax: 416 978 4317; E-mail: molly.shoichet@utoronto.ca

Cell Transplantation, Vol. 16, pp. 255-266, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Bone Marrow Mesenchymal Stem Cells From Healthy Donors and Sporadic Amyotrophic Lateral Sclerosis Patients

Ivana Ferrero,1 Letizia Mazzini,2 Deborah Rustichelli,1 Monica Gunetti,1 Katia Mareschi,1 Lucia Testa,2 Nicola Nasuelli,2 Gaia Donata Oggioni,2 and Franca Fagioli1

1Department of Pediatrics, "Regina Margherita Children's Hospital", University of Turin, Turin, Italy
2Department of Neurology, "Maggiore della Carità Hospital", University of Eastern Piedmont, Novara, Italy

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease lacking effective therapies. Cell replacement therapy has been suggested as a promising therapeutic approach for multiple neurodegenerative diseases, including motor neuron disease. We analyzed expanded mesenchymal stem cells (MSCs) isolated from sporadic ALS patients and compared them with MSCs isolated from healthy donors. MSCs were isolated from bone marrow by Percoll gradient and maintained in culture in MSC Medium until the third passage. Growth kinetics, immunophenotype, telomere length, and karyotype were evaluated during in vitro expansion. Osteogenic, adipogenic, chondrogenic, and neurogenic differentiation potential were also evaluated. No morphological differences were observed in the MSCs isolated from donors or patients. The cellular expansion potential of MSCs from donors and patients was slightly different. After three passages, the MSCs isolated from donors reached a cumulative population doubling higher than from patients but the difference was not statistically significant. No significant differences between donors or patients were observed in the immunophenotype analysis. No chromosomal alteration or evidence of cellular senescence was observed in any samples. Both donor and patient MSCs, after exposure to specific conditioning media, differentiated into adipocytes, osteoblasts, chondrocytes, and neuron-like cells. These results suggest that extensive in vitro expansion of patient MSCs does not involve any functional modification of the cells, including chromosomal alterations or cellular senescence. Hence, there is a good chance that MSCs might be used as a cell-based therapy for ALS patients.

Key words: Amyotrophic lateral sclerosis; Mesenchymal stem cells; In vitro expansion; Cellular therapy

Address correspondence to Ivana Ferrero, Sc.D., Department of Pediatrics, University of Turin, P.za Polonia 94, 10126 Turin, Italy. Tel: +39 011 3135566; Fax: +39 011 3135487; E-mail: ivana.ferrero@unito.it

Cell Transplantation, Vol. 16, pp. 267-278, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Implanted Mouse Bone Marrow-Derived Cells Reconstruct Layered Smooth Muscle Structures in Injured Urinary Bladders

Tetsuya Imamura, Yoshiaki Kinebuchi, Osamu Ishizuka, Satoshi Seki, Yasuhiko Igawa, and Osamu Nishizawa

Department of Urology, Shinshu University School of Medicine, Matsumoto, 390-8621, Japan

This study is a preliminary investigation to determine if bone marrow-derived cells, when implanted into freeze-injured urinary bladders, differentiate into smooth muscle cells and reconstruct smooth muscle layers. Bone marrow cells were harvested from femurs of male ICR mice and cultured in collagen-coated dishes for 7 days. After 5 days of culture, the cells were transfected with green fluorescent protein (GFP) genes for identification in recipient tissues. Three days prior to implantation, the posterior urinary bladder walls of female nude mice were injured with an iron bar refrigerated by dry ice. Seven days after the culture and 3 days after the injury, adherent, proliferating GFP-labeled bone marrow-derived cells (1.0 × 105 cells) were implanted into the injured regions. For controls, a cell-free solution was injected. At 14 days after implantation, the experimental urinary bladders were analyzed by histological, gene expression, and cystometric investigations. Just prior to implantation, the injured regions did not have any smooth muscle layers. After 14 days, the implanted cells surviving in the recipient tissues were detected with GFP antibody. The implanted regions had distinct smooth muscle layers composed of regenerated smooth muscle marker-positive cells. The implanted GFP-labeled cells differentiated into smooth muscle cells that formed into layers. The differentiated cells contacted each other within the implanted region as well as smooth muscle cells of the host. As a result, the reconstructed smooth muscle layers were integrated into the host tissues. Control mice injected with cell-free solution developed only few smooth muscle cells and no layers. Cystometric investigations showed that mice with implanted the cells developed bladder contractions similar to normal mice, whereas control mice did not. In summary, mouse bone marrow-derived cells can reconstruct layered smooth muscle structures in injured bladders to remediate urinary dysfunction.

Key words: Bone marrow cell; Urinary bladder; Smooth muscle; Cell differentiation; Mice

Address correspondence to Tetsuya Imamura, Department of Urology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan. Tel: +81-263-37-2661; Fax: +81-263-37-3082; E-mail: imatetu@sch.md.shinshu-u.ac.jp

Cell Transplantation, Vol. 16, pp. 279-290, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

MR Tracking of Magnetically Labeled Mesenchymal Stem Cells in Rat Kidneys With Acute Renal Failure

Jun-Hui Sun,1 Gao-Jun Teng,1 Sheng-Hong Ju,1 Zhan-Long Ma,1 Xiao-Li Mai,1 and Ming Ma2

1Laboratory of Molecular Imaging, Department of Radiology, Zhongda Hospital, Southeast University, Nanjing 210009, China
2Laboratory of Molecular and Biomolecular Electronics, Southeast University, Nanjing 210009, China

Stem cell transplantation is emerging as a potential treatment option for acute renal failure (ARF) because of its capability to regenerate tissues and organs. To better understand the mechanism of cell therapy, in vivo tracking cellular dynamics of the transplanted stem cells is needed. In the present study, in vivo monitored magnetically labeled mesenchymal stem cells (MSCs) were transplanted intravascularly into an ARF rat model using a conventional magnetic resonance imaging (MRI) system. Rat bone marrow MSCs were labeled with home synthesized Fe2O3-PLL, and labeled (n = 6) or unlabeled MSCs (n = 6) were injected into the renal arteries of the rats with ARF induced by the intramuscular injection of glycerol. Using the same technique, labeled MSCs were also injected into the rats assigned to a control group (n = 8). MR images of kidneys were obtained before injection of MSCs as well as immediately, 1, 3, 5, and 8 days afterwards. MR findings were analyzed and compared with histopathological and immunohistochemical results. These results showed that the rat MSCs were successfully labeled with the home synthesized Fe2O3-PLL. In both renal failure and intact rat models, the labeled MSCs demonstrated a loss of signal intensity in the renal cortex on T2*-weighted MR images, which was visible up to 8 days after transplantation. Histological analyses showed that most of the labeled MSCs that tested positive for Prussian blue staining were in glomerular capillaries, corresponding to the areas where a loss in signal intensity was observed in the MRI. A similar signal intensity decrease was not detected in the rats with unlabeled cells. These data demonstrate that the magnetically labeled MSCs in the rat model of ARF were successfully evaluated in vivo by a 1.5 T MRI system, showing that the mechanisms of stem cell therapy have great potential for future ARF treatment recipients.

Key words: Magnetic resonance imaging; Stem cells; Cell transplantation; Acute renal failure; Animal model

Address correspondence to Gao-Jun Teng, M.D., Department of Radiology, Zhong-Da Hospital, Southeast University, 87 Ding Jia Qiao Road, Nanjing 210009, China. Tel: 086 25 83272121; Fax: 086 25 83311083; E-mail: gjteng@vip.sina.com

Cell Transplantation, Vol. 16, pp. 291-301, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Multilineage Potential of Side Population Cells From Human Amnion Mesenchymal Layer

M. Kobayashi,1 T. Yakuwa,2 K. Sasaki,1 K. Sato,2 A. Kikuchi,1 I. Kamo,1 Y. Yokoyama,3 and N. Sakuragawa1

1Department of Regenerative Medicine, School of Allied Health Sciences, Kitasato University, Nishi-hashimoto, Kanagawa, Japan
2Department of Applied Life Science, Graduate School of Bioresource Sciences, Nihon University, Fujisawa, Kanagawa, Japan
3Stem Cell Processing, Inc., Nishi-hashimoto, Kanagawa, Japan

Side population (SP) cells were isolated by FACS from a human amnion mesenchymal cell (AMC) layer soon after enzyme treatment. The yield of SP cells from AMC layer (AMC-SP cells) was about 0.1-0.2%. AMC-SP cells grew well with cell doublings of 40-80 days of culture. FACS profiles and immunocytostaining showed that AMC-SP cells were composed of two different cells immunologically: HLA I-/II- and HLA I+/II-. Oct-3/4 was detected in the nucleus of AMC-SP cells, when the culture was examined at the third, sixth, and 10th passages. RT-PCR showed that AMC-SP cells expressed the Oct-4, Sox-2, and Rex-1 genes. Immunocytochemistry revealed that all AMC-SP cells were vimentin+, CK19+, and nestin+. In addition, flow cytometry analysis showed that SP cells had high expression of CD13, CD29, CD44, CD46, CD49b, CD49c, CD49e, CD59, CD140a, and CD166 but low expression of CD 49d, and CD51. No evidence of expression was obtained for CD34, CD45, CD49a, CD56, CD90, CD105, CD106, CD117, CD133, CD271, or Flk-1. Upon appropriate differentiation protocols, AMC-SP cells differentiated to several cell lineages such as neuroectodermal, osteogenic, chondrogenic, and adipogenic cells. These results indicate that AMC-SP cells have multilineage potential to several cell lineages with unique immunological characteristics such as HLA I-/II- or HLA I+/II-. AMC-SP cells should be of considerable value for regenerative medicine because they do not induce acute rejection after allotransplantation, they do not cause ethical issues, and there is no limit of supply.

Key words: Stem cells; Amnion mesenchymal cells; Side population cells; Multilineage potential; Mesenchymal cells

Address correspondence to Norio Sakuragawa, M.D., Ph.D., Department of Regenerative Medicine, School of Allied Health Sciences, Kitasato University, 5-4-30, Nishi-hashimoto, Kanagawa, Japan. Tel: 81-42-770-9330; Fax: 81-42-770-9330; E-mail: nor-sakura@sea.plala.or.jp

Cell Transplantation, Vol. 16, pp. 303-311, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Multipotent Menstrual Blood Stromal Stem Cells: Isolation, Characterization, and Differentiation

Amit N. Patel,1 Eulsoon Park,1 Michael Kuzman,1 Federico Benetti,2 Francisco J. Silva,2,3 and Julie G. Allickson4

1Center for Cardiac Cell Therapy-Heart Lung Esophageal Surgical Institute, University of Pittsburgh/UPMC/McGowan Institute of Regenerative Medicine, Pittsburgh, PA, USA
2Benetti Foundation, Rosario, Argentina
3NewStem Biosciences/PrimeGen Biotech, Irving, CA, USA
4Cryo-Cell International, Inc., Oldsmar, FL, USA

The stromal stem cell fraction of many tissues and organs has demonstrated to exhibit stem cell properties such as the capability of self-renewal and multipotency, allowing for multilineage differentiation. In this study, we characterize a population of stromal stem cells derived from menstrual blood (MenSCs). We demonstrate that MenSCs are easily expandable to clinical relevance and express multipotent markers such as Oct-4, SSEA-4, and c-kit at the molecular and cellular level. Moreover, we demonstrate the multipotency of MenSCs by directionally differentiating MenSCs into chondrogenic, adipogenic, osteogenic, neurogenic, and cardiogenic cell lineages. These studies demonstrate the plasticity of MenSCs for potential research in regenerative medicine.

Key words: Menstrual blood; Stromal stem cells; Multipotent markers; Differentiation

Address correspondence to Amit N. Patel, M.D., M.S., Director of Cardiac Cell Therapy, The Heart, Lung and Esophageal Surgery Institute, UPMC Presbyterian, McGowan Institute of Regenerative Medicine, 200 Lothrop Street, Suite C 700, Pittsburgh, PA 15213, USA. Tel: 412-648-6411; Fax: 412-647-8681; E-mail: patelan@upmc.edu or Julie G. Allickson, Ph.D., Vice President of Laboratory Operations and Research & Development, Cryo-Cell International, Inc., 700 Brooker Creek Blvd., Suite 1800, Oldsmar, FL 34677, USA. Tel: 813-749-2100; Fax: 813-749-2207; E-mail: jallickson@cryo-cell.com

Cell Transplantation, Vol. 16, pp. 313-323, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Murine Embryonic Stem Cell-Derived Hepatic Progenitor Cells Engraft in Recipient Livers With Limited Capacity of Liver Tissue Formation

Amar Deep Sharma,1* Tobias Cantz,2* Arndt Vogel,1 Axel Schambach,3 Dhivya Haridass,1 Markus Iken,1 Martina Bleidißel,2 Michael P. Manns,1 Hans R. Schöler,2 and Michael Ott1

1Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany
2Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Muenster, Germany
3Department of Experimental Hematology, Hannover Medical School, Hannover, Germany

Directed endodermal differentiation of murine embryonic stem (ES) cells gives rise to a subset of cells with a hepatic phenotype. Such ES cell-derived hepatic progenitor cells (ES-HPC) can acquire features of hepatocytes in vitro, but fail to form substantial hepatocyte clusters in vivo. In this study, we investigated whether this is due to inefficient engraftment or an immature phenotype of ES-HPC. ES cells engrafted into recipient livers of NOD/SCID mice with a similar efficacy as adult hepatocytes after 28 days. Because transplanted unpurified ES-HPC formed teratomas in the spleen and liver, we applied an albumin promoter/enhancer-driven reporter system to purify ES-HPC by cell sorting. RT-PCR analyses for hepatocyte-specific genes showed that the cells exhibited a hepatic phenotype, lacking the expression of the pluripotency marker Oct4, comparable to cells of day 11.5 embryos. Sorted ES-HPC derived from b-galactosidase transgenic ES cells were injected into fumaryl-acetoacetate-deficient (FAH-/-) SCID mice and analyzed after 8 to 12 weeks. Staining with X-gal solution revealed the presence of engrafted cells throughout the liver. However, immunostaining for the FAH protein indicated hepatocyte formation at a very low frequency, without evidence for large hepatocyte cluster formation. In conclusion, the limited repopulation capacity of ES-HPC is not caused by a failure of primary engraftment, but may be due to an immature hepatic phenotype of the transplanted ES-HPC.

Key words: Embryonic stem cells; Hepatic precursor cells; Cell transplantation; Metabolic liver disease

Address correspondence to Michael Ott, Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Carl-Neuberg-Str. 1, D-30623 Hannover, Germany. Tel: +49-511-220027-120; Fax: +49-511-220027-148; E-mail: ott-mhh@gmx.de

*Both authors contributed equally to this work.

Cell Transplantation, Vol. 16, pp. 325-335, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
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Differentiation of Murine Embryonic Stem Cells in Skeletal Muscles of Mice

Chai Tian,1 Yifan Lu,1 Re´nald Gilbert,1,2 and George Karpati1

1Neuromuscular Research Group, Montreal Neurological Institute, McGill University, Montreal, Quebec, Canada
2Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada

Possible myogenic differentiation of SSEA-1- and OCT-4-positive murine embryonic stem cells (ESCs) and embryoid bodies (EBs) was studied in vitro and in vivo. In vitro, ESC- or EB-derived ESCs (EBs/ESCs) showed only traces of Pax 3 and 7 expression by immunocytochemistry and Pax 3 expression by immunoblot. By RT-PCR, myogenic determinant molecules (myf5, myoD, and myogenin) were expressed by EBs/ESCs but not by ESCs. However, in such cultures, very rare contracting myotubes were still present. Suspensions of LacZ-labeled ESCs or EBs were injected into anterior tibialis muscles (ATM) of different cohorts of mice for the study of their survival and possible myogenic differentiation. The different cohorts of mice included isogenic adult 129/Sv, nonisogenic CD1 and mdx, as well as mdx immunosuppressed with 2.5 mg/kg daily injections of tacrolimus. Ten to 90 days postinjections, the injected ATM of nonisogenic mice did not contain cells positive for LacZ, SSEA-1, OCT-4, or embryonic myosin heavy chain. The ATM of intact mdx mice contained very rare examples of muscle fibers positive for dystrophin and/or embryonic myosin heavy chain. In the ATM of the isogenic normal and the immunosuppressed mdx mice, as expected, large teratomas developed containing the usual diverse cell types. In some teratomas of immunosuppressed mdx mice, small pockets of muscle fibers expressed dystrophin and myosin heavy chain. Our studies indicated that in muscles of animals nonisogenic with the used ESCs, only very rare ESCs survived with myogenic differentiation. These studies also indicated that ESCs will not undergo significant, selective, and preferential myogenic differentiation in vitro or in vivo in any of the models studied. It is probable that this strain of murine ESC requires some experimentally induced alteration of its gene expression profile to secure significant myogenicity and suppress tumorogenicity.

Key words: Murine embryonic stem cells; Embryoid bodies; Myogenic differentiation; mdx mice; Immunosuppression; Tacrolimus; Dystrophin expression; Teratoma formation

Address correspondence to Dr. George Karpati, Montreal Neurological Institute, 3801 University Street, Montreal, Quebec, Canada H3A 2B4. Tel: (514) 398-8528; Fax: (514) 398-8310; E-mail: george.karpati@mcgill.ca

Cell Transplantation, Vol. 16, pp. 337-350, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Inhibiting Myostatin With Follistatin Improves the Success of Myoblast Transplantation in Dystrophic Mice

Basma F. Benabdallah,1 Manaf Bouchentouf,1 Joel Rousseau,1 Pascal Bigey,2 Annick Michaud,1 Pierre Chapdelaine,3 Daniel Scherman,2 and Jacques P. Tremblay1

1Génétique humaine, Centre de Recherche du CHUL, Québec, Canada
2Inserm, U640, CNRS, UMR8151, Rene´ Descartes Paris 5 University, Faculté des Sciences Pharmaceutiques et Biologiques, Chemical and Genetic Pharmacology Laboratory, Ecole Nationale Suprieure de Chimie de Paris, Paris, France
3Ontogénie et reproduction, Centre de Recherche du CHUL, Québec, Canada

Duchenne muscular dystrophy is a recessive disease due to a mutation in the dystrophin gene. Myoblast transplantation permits to introduce the dystrophin gene in dystrophic muscle fibers. However, the success of this approach is reduced by the short duration of the regeneration following the transplantation, which reduces the number of hybrid fibers. Our aim was to verify whether the success of the myoblast transplantation is enhanced by blocking the myostatin signal with an antagonist, follistatin. Three different approaches were studied to overexpress follistatin in the muscles of mdx mice transplanted with myoblasts. First, transgenic follistatin/mdx mice were generated; second, a follistatin plasmid was electroporated in mdx muscles, and finally, follistatin was induced in mdx mice muscles by a treatment with a histone deacetylase inhibitor. The three approaches improved the success of the myoblast transplantation. Moreover, fiber hypertrophy was also observed in all muscles, demonstrating that myostatin inhibition by follistatin is a good method to improve myoblast transplantation and muscle function. Myostatin inhibition by follistatin in combination with myoblast transplantation is thus a promising novel therapeutic approach for the treatment of muscle wasting in diseases such as Duchenne muscular dystrophy.

Key words: Duchenne muscular dystrophy; Myoblast transplantation; Myostatin; Follistatin

Address correspondence to Jacques P. Tremblay, Ph.D., Unité de recherche en Génétique humaine, Centre de recherche de l'Université Laval, 2705, boulevard Laurier, RC-9300, Québec (Prov. Québec), Canada G1V 4G2. Tel: (418) 654-2186; Fax: (418) 654-2207; E-mail: Jacques-P.Tremblay@crchul.ulaval.ca

Cell Transplantation, Vol. 16, pp. 351-360, 2008
0963-6897/08 $90.00 + 00
E-ISSN 1555-3892
Copyright © 2008 Cognizant Comm. Corp.
Printed in the USA. All rights reserved.

Cultured Melanocytes: From Skin Biopsy to Transplantation

Deepa Ghosh, Sudheer Shenoy, and Pushpa Kuchroo

Tissue Engineering Group, Reliance Life Sciences Pvt. Ltd., Navi Mumbai, India

Restoration of cutaneous pigmentation has been achieved in stable vitiligo by autologous melanocyte transplantation. This study was aimed to develop a methodology to deliver melanocytes to vitiliginous area following their processing and culture in a centralized facility. Here we report a methodology to culture melanocytes on carrier films, transport the cells, and graft them on vitiliginous areas. The salient features of this study include: 1) development of polylactic acid (PLA) films that support melanocyte attachment, growth, and delivery; 2) establish transport conditions for skin biopsies from hospitals; 3) establish transport conditions for cultured cells from cell processing center to hospitals. Results suggest that PLA films could serve as carriers for melanocytes during transport. "pside-down"application of the graft results in the migration of cells from the films into the dermabraded area. The transport conditions ensure cell viability for 96 h. This system could help clinicians, who do not have access to cell culture facilities, transplant cultured melanocytes in a cost-effective manner.

Key words: Melanocytes; Transplantation; Treatment; Vitiligo

Address correspondence to Deepa Ghosh, Ph.D., Tissue Engineering Group, Reliance Life Sciences Pvt Ltd., Dhirubhai Ambani Life Sciences Centre, Thane Belapur Road, Rabale, Navi Mumbai 400 701, India. Tel: +1-22-39118428; Fax: +1-22-39115439; E-mail: deepa_ghosh@relbio.com