Cell Medicine 4(2) Abstracts

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Cell Medicine, Vol. 4, pp. 55–63, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X639306
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Nestin Overexpression Precedes Caspase-3 Upregulation in Rats Exposed to Controlled Cortical Impact Traumatic Brain Injury

Yuji Kaneko,* Naoki Tajiri,* SeongJin Yu,* Takuro Hayashi,* Christine E. Stahl,† Eunkyung Bae,* Humberto Mestre,* Nicholas Franzese,* Antonio Rodrigues Jr.,* Maria C. Rodrigues,* Hiroto Ishikawa,* Kazutaka Shinozuka,* Whitney Hethorn,* Nathan Weinbren,* Loren E. Glover,* Jun Tan,‡§ Anilkumar Harapanahalli Achyuta,¶ Harry van Loveren,* Paul R. Sanberg,* Sundaram Shivsankar,¶ and Cesar V. Borlongan*

*Department of Neurosurgery and Brain Repair, University of South Florida, Tampa, FL, USA
†Department of Aerospace Medicine, MacDill Air Force Base, Tampa, FL, USA
‡James A. Haley Veterans’ Administration Hospital, Tampa, FL, USA
§Rashid Laboratory for Developmental Neurobiology, Department of Psychiatry and Behavioral Neurosciences, University of South Florida, Tampa, FL, USA
¶Draper Laboratory, Bioengineering Center, Tampa, FL, USA

Our understanding of biological mechanisms and treatment options for traumatic brain injury (TBI) is limited. Here, we employed quantitative real-time PCR (QRT-PCR) and immunohistochemical analyses to determine the dynamic expression of cell proliferation and apoptosis in an effort to provide insights into the therapeutic window for developing regenerative strategies for TBI. For this purpose, young adult Sprague–Dawley rats were subjected to experimental TBI using a controlled cortical impactor and then euthanized 1–48 h after TBI for QRT-PCR and immunohistochemistry. QRT-PCR revealed that brains from TBI-exposed rats initially displayed nestin mRNA expression that modestly increased as early as 1 h post-TBI, then significantly peaked at 8 h, but thereafter reverted to pre-TBI levels. On the other hand, caspase-3 mRNA expression was slightly elevated at 8 h post-TBI, which did not become significantly upregulated until 48 h. Immunofluorescent microscopy revealed a significant surge in nestin-immunoreactive cells in the cortex, corpus callosum, and subventricular zone at 24 h post-TBI, whereas a significant increase in the number of active caspase-3-immunoreactive cells was only found in the cortex and not until 48 h. These results suggest that the injured brain attempts to repair itself via cell proliferation immediately after TBI but this endogenous regenerative mechanism is not sufficient to abrogate the secondary apoptotic cell death. Treatment strategies designed to amplify cell proliferation and to prevent apoptosis are likely to exert maximal benefits when initiated at the acute phase of TBI.

Key words: Cell proliferation; Neurogenesis; Apoptosis; Head trauma; Nestin; Caspase-3

Received July 14, 2011; final acceptance March 27, 2012. Online prepub date: April 10, 2012.
Address correspondence to Cesar V. Borlongan, Ph.D., Professor and Vice-Chairman for Research, Department of Neurosurgery and Brain Repair, University of South Florida, Tampa, FL 33612, USA. Tel: +1 813 974 3154; Fax: +1 813 974 3078; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 65–76, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X647172
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

The Effect of CXCR4 Overexpression on Mesenchymal Stem Cell Transplantation in Ischemic Stroke

Oh Young Bang,* Kyung Sil Jin,† Mi Na Hwang,* Ho Young Kang,* Byoung Joon Kim,* Sang Jin Lee,‡ Sangmee Kang,§ Yu Kyeong Hwang,§ Jong Seong Ahn,¶ and Ki Woong Sung†

*Department of Neurology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Korea
†Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Korea
‡Genitourinary Cancer Branch, National Cancer Center, Goyang, South Korea
§Cancer Therapeutics Team II, Mogam Biotechnology Research Institute, Yongin, South Korea
¶Cell Therapy Division, GCLabCell Corp., Yongin, South Korea

There is no doubt that the therapeutic efficacy of mesenchymal stem cells (MSCs) needs improvement. SDF-1 (chemokine for MSC homing) and its receptor CXCR4 play a critical role in the migration of MSCs in ischemia. We investigated the effects of the therapeutic application of MSCs transfected to overexpress CXCR4 using an adenoviral construct in the rat stroke model. Both flow cytometry and Western blot analysis indicated that the level of CXCR4 expression was low in naive hMSCs but was consistently high in CXCR4-hMSCs. In vivo migration test using the transwell system showed that the degree of migration was increased in CXCR4-hMSCs compared with the naive hMSCs and was completely blocked by treatment with AMD3100, an antagonist of the CXCR4 receptor. Compared with rats that received naive MSCs, behavioral recovery was more pronounced in rats that received CXCR4-hMSCs (p = 0.023). An immunohistochemistry study using human nuclear antibody (NuMA) showed that the migration of hMSCs in the ischemic boundary zone was increased after 3 days of injection of CXCR4-hMSCs compared with after injection of naive hMSCs. In addition, polymerase chain reaction was performed to assess the biodistribution of human-specific DNA outside the brain after intravenous injection of hMSCs. The expression of human-specific DNA was increased in the lungs of rats receiving naive MSCs, whereas the human-specific DNA expression was increased in the brain of rats receiving CXCR4-hMSCs. Our results indicate that MSCs transfected with the CXCR4 gene expression cassette may be useful in the treatment of cerebral infarction and may represent a new strategy to enhance the efficacy of MSC therapy.

Key words: Cerebral infarction; Mesenchymal stem cells; Stem cell; SDF-1; Chemokine; CXCR4; Adenovirus

Received September 13, 2011; final acceptance March 15, 2012. Online prepub date: May 15, 2012.
Address correspondence to Ki Woong Sung, M.D., Ph.D., Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul 135-710, South Korea. Tel: +82-2-3410-3539; Fax: +82-2-3410-0043; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 77–84, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X653337
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Use of Magnetocapsules for In Vivo Visualization and Enhanced Survival of Xenogeneic HepG2 Cell Transplants

Thomas W. Link,*†§ Dian R. Arifin,*§ Christopher M. Long,† Piotr Walczak,*§ Naser Muja,*§ Aravind Arepally,¶#** and Jeff W.M. Bulte*†‡§

*Russell H. Morgan Department of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
†Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
‡Department of Chemical and Biomolecular Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
§Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering,The Johns Hopkins University School of Medicine, Baltimore, MD, USA
¶Division of Interventional Radiology, Piedmont Hospital, Atlanta, GA, USA
#Department of Radiology, The Johns Hopkins Medical Institutes, Baltimore, MD, USA
**Department of Surgery, The Johns Hopkins Medical Institutes, Baltimore, MD, USA

Hepatocyte transplantation is currently being considered as a new paradigm for treatment of fulminant liver failure. Xeno- and allotransplantation studies have shown considerable success, but the long-term survival and immunorejection of engrafted cells need to be further evaluated. Using novel alginate–protamine sulfate–alginate microcapsules, we have coencapsulated luciferase-expressing HepG2 human hepatocytes with superparamagnetic iron oxide nanoparticles to create magnetocapsules that are visible on MRI as discrete hypointensities. Magnetoencapsulated cells survive and secrete albumin for at least 5 weeks in vitro. When transplanted intraperitoneally in immunocompetent mice, encapsulated hepatocytes survive for at least 4 weeks as determined using bioluminescent imaging, which is in stark contrast to naked, unencapsulated hepatocytes that died within several days after transplantation. However, in vivo human albumin secretion did not follow the time course of magnetoencapsulated cell survival, with plasma levels returning to baseline values already at 1 week posttransplantation. The present results demonstrate that encapsulation can dramatically prolong survival of xenotransplanted hepatocytes, leading to sustained albumin secretion with a duration that may be long enough for use as a temporary therapeutic bridge to liver transplantation.

Key words: Cell transplantation; Fulminant liver failure; Magnetic resonance imaging; Iron nanoparticle contrast agent; Bioluminescent imaging

Received February 29, 2012; final acceptance July 27, 2012. Online prepub date: August 27, 2012.
Address correspondence to Jeff W.M. Bulte, Ph.D., Russell H. Morgan Department of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, 217 Traylor Building, 720 Rutland Avenue, Baltimore, MD 21205, USA. Tel.: +1 443 287 0996;Fax: +1 443 287 6730; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 85–98, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X647136
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Improved Hepatocyte Engraftment After Portal Vein Occlusion in LDL Receptor-Deficient WHHL Rabbits and Lentiviral-Mediated Phenotypic Correction In Vitro

Sylvie Goulinet-Mainot,*1 Hadrien Tranchart,*1 Marie-Thérèse Groyer-Picard,* Panagiotis Lainas,*† Papa Saloum Diop,*† Delphine Holopherne,‡ Patrick Gonin,§ Karim Benihoud,¶ Nathalie Ba,# Olivier Gauthier,‡ Dominique Franco,*† Catherine Guettier,** Danièle Pariente,*†† Anne Weber,* Ibrahim Dagher,*† and Tuan Huy Nguyen‡‡

*INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France
†Department of General Surgery, Univ. Paris-Sud, Antoine Béclère Hospital, Clamart, France
‡Department of Animal Surgery, Veterinary School of Nantes, Nantes, France
§IFR 54, Service Commun d’Expérimentation Animale, Institut Gustave Roussy, Villejuif, France
¶CNRS UMR 8203, Institut Gustave Roussy, Villejuif, France
#IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France
**Department of Pathology, Bicêtre Hospital, Le Kremlin-Bicêtre, France
††Department of Pediatric Radiology, Bicêtre Hospital, Le Kremlin-Bicêtre, France
‡‡INSERM U1064, CHU Hôtel Dieu, Université de Nantes, Nantes, France

Innovative cell-based therapies are considered as alternatives to liver transplantation. Recent progress in lentivirusmediated hepatocyte transduction has renewed interest in cell therapy for the treatment of inherited liver diseases. However, hepatocyte transplantation is still hampered by inefficient hepatocyte engraftment. We previously showed that partial portal vein embolization (PVE) improved hepatocyte engraftment in a nonhuman primate model. We developed here an ex vivo approach based on PVE and lentiviral-mediated transduction of hepatocytes from normal (New Zealand White, NZW) and Watanabe heritable hyperlipidemic (WHHL) rabbits: the large animal model of familial hypercholesterolemia type IIa (FH). FH is a life-threatening human inherited autosomal disease caused by a mutation in the low-density lipoprotein receptor (LDLR) gene, which leads to severe hypercholesterolemia and premature coronary heart disease. Rabbit hepatocytes were isolated from the resected left liver lobe, and the portal branches of the median lobes were embolized with Histoacryl® glue under radiologic guidance. NZW and WHHL hepatocytes were each labeled with Hoechst dye or transduced with lentivirus expressing GFP under the control of a liver-specific promoter (mTTR, a modified murine transthyretin promoter) and were then immediately transplanted back into donor animals. In our conditions, 65–70% of the NZW and WHHL hepatocytes were transduced. Liver repopulation after transplantation with the Hoechst-labeled hepatocytes was 3.5 ± 2%. It was 1.4 ± 0.6% after transplantation with either the transduced NZW hepatocytes or the transduced WHHL hepatocytes, which was close to that obtained with Hoechst-labeled cells, given the mean transduction efficacy. Transgene expression persisted for at least 8 weeks posttransplantation. Transduction of WHHL hepatocytes with an LDLR-encoding vector resulted in phenotypic correction in vitro as assessed by internalization of fluorescent LDL ligands. In conclusion, our results have applications for the treatment of inherited metabolic liver diseases, such as FH, by transplantation of lentivirally transduced hepatocytes.

Key words: Hepatocyte transplantation; Liver; Rabbit; Lentiviral vector; Portal vein embolization; Familial hypercholesterolemia

Received September 19, 2011; final acceptance April 26, 2012. Online prepub date: May 8, 2012.
1These authors provide equal contribution to this work.
Address correspondence to Ibrahim Dagher, M.D., Ph.D., Department of General Surgery, Antoine Béclère Hospital, 157 rue de la Porte de Trivaux, 92141 Clamart cedex, France. Tel: +33 (0) 6 12 75 13 14; Fax: +33 (0) 1 45 37 49 78; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 99–107, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X647226
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Behavior of Human Articular Chondrocytes During In Vivo Culture in Closed, Permeable Chambers

Iñigo Martinez-Zubiaurre,* Tuija Annala,† and Martin Polacek*‡

*Orthopaedic Surgery Department, Institute of Clinical Medicine, University of Tromsø, Tromsø, Norway
†Scaffdex Oy, Tampere, Finland
‡Orthopaedic Surgery Department, University Hospital of North Norway, Tromsø, Norway

The exact contribution of transplanted chondrocytes for cartilage tissue repair prior expansion in monolayer cultures remains undetermined. At our laboratory, we have created a new permeable chamber to study the chondrogenesis of dedifferentiated cells implanted ectopically in a closed and controlled environment. The behavior of chondrocytes has been studied in settings frequently used in clinical approaches during transplantation, namely injection of autologous chondrocyte cells in suspension (ACI), cells soaked in collagen membranes (MACI), and cells applied in a polymer gel (fibrin). As controls, we have tested the redifferentiation of chondrocytes in cell aggregates, and we have checked the proper functionality of chambers both in vitro and in vivo. After retrieval, firmed tissue-like shapes were recovered only from chambers containing cells seeded in membranes. Histomorphological, immunohistochemical, and ultrastructural analyses revealed synthesis of fibrous-like tissue, characterized by low-density collagen fibers, low collagen type II, abundant collagen type I, and low amounts of proteoglycans. Additionally, neither the collagen membranes nor the fibrin gel was reabsorbed by cells. In summary, our results show that the newly developed permeable chambers function correctly, allowing proper cell feeding and preventing cell leakage or host cell invasion. Additionally, our results suggest that, under these circumstances, chondrocytes are not able to orchestrate formation of hyaline cartilage and have little capacity to degrade artificial membranes or carrier gels such as fibrin. These are interesting observations that should be considered for understanding what role the transplanted chondrocytes play during restoration of articular cartilage after implantation.

Key words: Closed permeable chambers; Cartilage; Chondrogenesis; Collagen membranes

Received May 30, 2011; final acceptance June 3, 2012. Online prepub date: August 7, 2012.
Address correspondence to Inigo Martinez, Bone and Joint Research Group, Orthopaedic Surgery Department, Institute of Clinical Medicine, University of Tromsø, 9037 Tromsø, Norway. Tel.: +4777644686; Fax: +4777627164; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it