Cell Medicine 4(1) Abstracts

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Cell Medicine, Vol. 4, pp. 1–11, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X647217
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells1

Danielle D. Carrade,* Michael W. Lame,† Michael S. Kent,‡ Kaitlin C. Clark,* Naomi J. Walker,* and Dori L. Borjesson*

*Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis, CA, USA
†Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA, USA
‡Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, CA, USA

Mesenchymal stem cells (MSCs) derived from bone marrow (BM), adipose tissue (AT), umbilical cord blood (CB), and umbilical cord tissue (CT) are increasingly being used to treat equine inflammatory and degenerative lesions. MSCs modulate the immune system in part through mediator secretion. Animal species and MSC tissue of origin are both important determinants of MSC function. In spite of widespread clinical use, how equine MSCs function to heal tissues is fully unknown. In this study, MSCs derived from BM, AT, CB, and CT were compared for their ability to inhibit lymphocyte proliferation and secrete mediators in response to activation. Five MSC lines from each tissue were isolated. Lymphocyte proliferation was assessed in a mixed leukocyte reaction, and mediator secretion was determined by ELISA. Regardless of tissue of origin, quiescent MSCs did not alter lymphocyte proliferation or secrete mediators, except for transforming growth factor-β (TGF-β1). When stimulated, MSCs of all tissue types decreased lymphocyte proliferation, increased prostaglandin (PGE2) and interleukin-6 (IL-6) secretion, and decreased production of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). BM-MSCs and CB-MSCs also produced nitric oxide (NO), while AT-MSCs and CT-MSCs did not. Equine MSCs did not produce indoleamine 2,3-dioxygenase (IDO). These data suggest that activated equine MSCs derived from BM, AT, CT, and CB secrete high concentration of mediators and are similar to MSCs from rodents and humans in their immunomodulatory profiles. These findings have implication for the treatment of inflammatory lesions dominated by activated lymphocytes and TNF-α and IFN-γ in vivo.

Key words: Equine; Mesenchymal stem cells; Immunomodulation; Lymphocytes; Bone marrow; Umbilical cord blood; Adipose and umbilical cord tissue

Received June 23, 2011; final acceptance May 10, 2012. Online prepub date: July 15, 2012.
1This work was presented as an abstract at North American Veterinary Regenerative Medicine Conference, June 2–4, 2011, Lexington, KY.

Address correspondence to Dr. Dori L. Borjesson, Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA. Tel.: +1 530 754 5202; Fax: +1 530 752 3349; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 13–21, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X653319
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Comparison of Gingiva, Dental Pulp, and Periodontal Ligament Cells From the Standpoint of Mesenchymal Stem Cell Properties

Koji Otabe,* Takeshi Muneta,*† Nobuyuki Kawashima,‡ Hideaki Suda,†‡ Kunikazu Tsuji,† and Ichiro Sekiya§

*Orthopedic Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
†Global Center of Excellence Program for International Research Center for Molecular Science in Tooth and Bone Disease, Tokyo Medical and Dental University, Tokyo, Japan
‡Pulp Biology and Endodontics, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
§Cartilage Regeneration, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan

The specific properties of mesenchymal stem cells (MSCs) in oral tissues still remain unknown though their existence has been previously reported. We collected gingiva, dental pulp, and periodontal ligament tissues from removed teeth and isolated MSCs. These MSCs were compared in terms of their yields per tooth, surface epitopes, and differentiation potentials by patient-matched analysis. For in vivo calcification analysis, rat gingival and dental pulp cells mounted on b-tricalcium phospateTCP were transplanted into the perivertebral muscle of rats for 6 weeks. Gingival cells and dental pulp cells showed higher yield per tooth than periodontal ligament cells (n = 6, p < 0.05). Yields of periodontal ligament cells were too low for further analysis. Gingival and dental pulp cells expressed MSC markers such as CD44, CD90, and CD166. Gingival and dental pulp cells obtained phenotypes of chondrocytes and adipocytes in vitro. Approximately 60% of the colonies of gingival cells and 40% of the colonies of dental pulp cells were positively stained with alizarin red in vitro, and both gingival and dental pulp cells were calcified in vivo. We clarified properties of MSCs derived from removed teeth. We could obtain a high yield of MSCs with osteogenic potential from gingiva and dental pulp. These results indicate that gingiva and dental pulp are putative cell sources for hard tissue regeneration.

Key words: Mesenchymal stem cells (MSCs); Gingiva; Dental pulp; Periodontal ligament; Yields; Differentiation

Received January 26, 2012; June 6, 2012. Online prepub date: August 10, 2012.
Address correspondence correspondence to Ichiro Sekiya M.D., Ph.D., Section of Cartilage Regeneration, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan. Tel: +81-3-5803-4675; Fax: +81-3-5803-0266; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 23–32, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X639324
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Epiphyseal Chondroprogenitors Provide a Stable Cell Source for Cartilage Cell Therapy

Salim Darwiche,*† Corinne Scaletta,* Wassim Raffoul,* Dominique P. Pioletti,† and Lee Ann Applegate*

*Regenerative Therapy Unit, Service of Plastic and Reconstructive Surgery, University Hospital of Lausanne, Switzerland
†Laboratory of Biomechanical Orthopedics, Ecole Polytechnique Federale de Lausanne, Switzerland

Articular cartilage regeneration poses particularly tough challenges for implementing cell-based therapies. Many cell types have been investigated looking for a balanced combination of responsiveness and stability, yet techniques are still far from defining a gold standard. The work presented focuses on the reliable expansion and characterization of a clinical grade human epiphyseal chondroprogenitor (ECP) cell bank from a single tissue donation. A parental human ECP cell bank was established, which provides the seed material for master and working cell banks. ECPs were investigated at both low and high cumulative population doublings looking at morphology, monolayer expansion kinetics, resistance to cryogenic shock, colony-forming efficiency, and cell surface markers. Three-dimensional micropellet assays were used to determine spontaneous extracellular matrix deposition at varying population doublings and monolayer 2D differentiation studies were undertaken to assess the propensity for commitment into other lineages and their stability. ECPs exhibited remarkable homogeneity in expansion with a steady proliferative potential averaging three population doublings over 8 days. Surface marker analysis revealed no detectable contaminating subpopulations or population enrichment during prolonged culture periods. Despite a slight reduction in Sox9 expression levels at higher population doublings in monolayer, nuclear localization was equivalent both in monolayer and in micropellet format. Equally, ECPs were capable of depositing glycosaminoglycans and producing aggrecan, collagen I, and collagen II in 3D pellets both at low and high population doublings indicating a stable spontaneous chondrogenic potential. Osteogenic induction was differentially restricted in low and high population doublings as observed by Von Kossa staining of calcified matrix, with a notable collagen X, MMP13, and ADAMTS5 downregulation. Rare adipogenic induction was seen as evidenced by cytoplasmic lipid accumulation detectable by Oil Red O staining. These findings highlight the reliability, stability, and responsiveness of ECPs over prolonged culture, making them ideal candidates in defining novel strategies for cartilage regeneration.

Key words: Chondrocytes; Progenitor cells; Articular cartilage regeneration; Cell banking

Received November 3, 2011; final acceptance April 12, 2012. Online prepub date: May 8, 2012.
Address correspondence to Professor Lee Ann Applegate, Regenerative Therapy Unit, Service of Plastic and Reconstructive Surgery, University Hospital of Lausanne, Pavillon 3, CH-1011 Lausanne, Switzerland. Tel: +41 21 314 35 10; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 33–43, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X653328
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Bioartificial Renal Epithelial Cell System (BRECS): A Compact, Cryopreservable Extracorporeal Renal Replacement Device

Deborah A. Buffington,* Christopher J. Pino,* Lijun Chen,* Angela J. Westover,* Gretchen Hageman,* and H. David Humes*†‡

*Innovative BioTherapies, Inc., Ann Arbor, MI, USA
†Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI, USA
‡CytoPherx, Inc., Ann Arbor, MI, USA

Renal cell therapy has shown clinical efficacy in the treatment of acute renal failure (ARF) and promise for treatment of end-stage renal disease (ESRD) by supplementing conventional small solute clearance (hemodialysis or hemofiltration) with endocrine and metabolic function provided by cells maintained in an extracorporeal circuit. A major obstacle in the widespread adoption of this therapeutic approach is the lack of a cryopreservable system to enable distribution, storage, and therapeutic use at point of care facilities. This report details the design, fabrication, and assessment of a Bioartificial Renal Epithelial Cell System (BRECS), the first all-in-one culture vessel, cryostorage device, and cell therapy delivery system. The BRECS was loaded with up to 20 cell-seeded porous disks, which were maintained by perfusion culture. Once cells reached over 5 × 106 cells/disk for a total therapeutic dose of approximately 108 cells, the BRECS was cryopreserved for storage at −80°C or −140°C. The BRECS was rapidly thawed, and perfusion culture was resumed. Near precryopreservation values of cell viability, metabolic activity, and differentiated phenotype of functional renal cells were confirmed postreconstitution. This technology could be extended to administer other cell-based therapies where metabolic, regulatory, or secretion functions can be leveraged in an immunoisolated extracorporeal circuit.

Key words: Extracorporeal cell therapy; Progenitor; Cryopreservation; Bioreactor; Acute renal failure; End-stage renal disease

Received October 21, 2011; final acceptance May 31, 2012. Online prepub date: August 10, 2012.
Address correspondence to Dr. David Humes, Innovative BioTherapies, Inc., 401 W. Morgan Rd., Ann Arbor, MI 48108, USA. Tel.: +1 734 213 8350; Fax +1 734 213 8920; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 45–54, 2012
2155-1790/12 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X639315
Copyright © 2012 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Adequate Time Window and Environmental Factors Supporting Retinal Graft Cell Survival in rd Mice

Michiko Mandai, Kohei Homma, Satoshi Okamoto, Chikako Yamada, Akane Nomori, and Masayo Takahashi

Laboratory for Retinal Regeneration, Riken Kobe CDB, Kobe, Hyogo, Japan

Postnatal photoreceptor cells can be integrated into the wild-type adult retina in mice, and retinal transplantation is now one therapeutic option for retinal degenerative diseases when photoreceptor degeneration is the primary cause of the disease. The aim of this study was to specify the optimal time window during the course of retinal degeneration and to modulate the host and/or graft environment for a successful transplantation. We first studied the background features of the mice with phosphodiesterase 6b (PDE6b) gene mutation (rd; C3H/Hej) and found that the infiltration of microglia and glial fibrillary acidic protein (GFAP) expression once increased at the peak of rod death (~2–3 weeks of age) but then reduced for a following period until gliosis began to take place with enhanced GFAP expression (~8 weeks of age). The postnatal retinal cells (p4–p7) were successfully transplanted during this period with neurite extension into the host retina. In later transplantations (6 or 8 weeks of age), graft cells survived better in the presence of chondroitinase ABC (ChABC), which digests chondroitin sulfate proteoglycan (CSPG), an essential component of gliosis. In contrast, in earlier transplantations (4 weeks of age), graft cells survived better in the presence of valproic acid (VPA), a neural differentiating reagent, or glatiramer acetate, an immune modulator. These suggest that, immediately after the outer nuclear layer (ONL) degeneration, an inflammatory reaction may be easily induced but the host neurons may be more able to accept donor cells in the presence of neural differentiating factor. These results will help optimize transplantation conditions when we consider clinical application.

Key words: Photoreceptors; Transplantation; Gliosis; Microglia; Glatiramer acetate; Valproic acid

Received June 15, 2011; final acceptance March 30, 2012. Online prepub date: April 20, 2012.
Address correspondence to Michiko Mandai, Laboratory for Retinal Regeneration, Riken Kobe CDB, 2-2-3 Nubatihuna-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan. Tel: +81-78-306-3302; Fax: +81-88-306-3303; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it