Cell Medicine 4(3) Abstracts

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Cell Medicine, Vol. 4, pp. 109–123, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X658927
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Simple Machine Perfusion Significantly Enhances Hepatocyte Yields of Ischemic and Fresh Rat Livers

Maria-Louisa Izamis,* Candice Calhoun,* Basak E. Uygun,* Maria Angela Guzzardi,* Gavrielle Price,* Martha Luitje,* Nima Saeidi,* Martin L. Yarmush,*† and Korkut Uygun*

*Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, MA, USA
†Department of Biomedical Engineering, Rutgers University, Piscataway, NJ, USA

The scarcity of viable hepatocytes is a significant bottleneck in cell transplantation, drug discovery, toxicology, tissue engineering, and bioartificial assist devices, where trillions of high-functioning hepatocytes are needed annually. We took the novel approach of using machine perfusion to maximize cell recovery, specifically from uncontrolled cardiac death donors, the largest source of disqualified donor organs. In a rat model, we developed a simple 3-h room temperature (20 ± 2°C) machine perfusion protocol to treat nonpremedicated livers exposed to 1 h of warm (34°C) ischemia. Treated ischemic livers were compared to fresh, fresh-treated, and untreated ischemic livers using viable hepatocyte yields and in vitro performance as quantitative endpoints. Perfusion treatment resulted in both a 25-fold increase in viable hepatocytes from ischemic livers and a 40% increase from fresh livers. While cell morphology and function in suspension and plate cultures of untreated warm ischemic cells was significantly impaired, treated warm ischemic cells were indistinguishable from fresh hepatocytes. Furthermore, a strong linear correlation between tissue ATP and cell yield enabled accurate evaluation of the extent of perfusion recovery. Maximal recovery of warm ischemic liver ATP content appears to be correlated with optimal flow through the microvasculature. These data demonstrate that the inclusion of a simple perfusion-preconditioning step can significantly increase the efficiency of functional hepatocyte yields and the number of donor livers that can be gainfully utilized.

Key words: Disqualified cardiac death (DCD); Nonheart beating donor (NHBD); Ischemia; Adenosine triphospate (ATP); Cell isolation

Received February 27, 2012; final acceptance October 23, 2012. Online prepub date: November 27, 2012.
Address correspondence to Korkut Uygun, Ph.D., 51 Blossom St., Boston, MA 02114, USA. Tel: +1-617-371-4881; Fax: +1-617-573-9471; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 125–147, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X658918
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Human Decidua-Derived Mesenchymal Cells Are a Promising Source for the Generation and Cell Banking of Human Induced Pluripotent Stem Cells

Tomoko Shofuda,* Daisuke Kanematsu,† Hayato Fukusumi,† Atsuyo Yamamoto,* Yohei Bamba,‡ Sumiko Yoshitatsu,§ Hiroshi Suemizu,¶ Masato Nakamura,¶# Yoshikazu Sugimoto,** Miho Kusuda Furue,†† Arihiro Kohara,‡‡ Wado Akamatsu,‡ Yohei Okada,‡§§ Hideyuki Okano,‡ Mami Yamasaki,¶¶##*** and Yonehiro Kanemura†##

*Division of Stem Cell Research, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Chuo-ku, Osaka, Japan
†Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan
‡Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan
§Department of Plastic Surgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan
¶Biomedical Research Department, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Japan
#Department of Pathology and Regenerative Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan
**Division of Chemotherapy, Faculty of Pharmacy, Keio University, Minato-ku, Tokyo, Japan
††Laboratory of Stem Cell Cultures, Laboratory of Cell Cultures, Department of Disease Bioresources Research, National Institute of Biomedical Innovation, Ibaraki, Osaka, Japan
‡‡JCRB Cell Bank, Laboratory of Cell Cultures, Research on Disease Bioresources, National Institute of Biomedical Innovation, Osaka, Japan
§§Kanrinmaru-Project, School of Medicine, Keio University, Tokyo, Japan
¶¶Division of Molecular Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan
##Department of Neurosurgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan
***Department of Pediatric Neurosurgery, Takatsuki General Hospital, Takatsuki, Osaka, Japan

Placental tissue is a biomaterial with remarkable potential for use in regenerative medicine. It has a three-layer structure derived from the fetus (amnion and chorion) and the mother (decidua), and it contains huge numbers of cells. Moreover, placental tissue can be collected without any physical danger to the donor and can be matched with a variety of HLA types. The decidua-derived mesenchymal cells (DMCs) are highly proliferative fibroblast-like cells that express a similar pattern of CD antigens as bone marrow-derived mesenchymal cells (BM-MSCs). Here we demonstrated that induced pluripotent stem (iPS) cells could be efficiently generated from DMCs by retroviral transfer of reprogramming factor genes. DMC-hiPS cells showed equivalent characteristics to human embryonic stem cells (hESCs) in colony morphology, global gene expression profile (including human pluripotent stem cell markers), DNA methylation status of the OCT3/4 and NANOG promoters, and ability to differentiate into components of the three germ layers in vitro and in vivo. The RNA expression of XIST and the methylation status of its promoter region suggested that DMC-iPSCs, when maintained undifferentiated and pluripotent, had three distinct states: (1) complete X-chromosome reactivation, (2) one inactive X-chromosome, or (3) an epigenetic aberration. Because DMCs are derived from the maternal portion of the placenta, they can be collected with the full consent of the adult donor and have considerable ethical advantages for cell banking and the subsequent generation of human iPS cells for regenerative applications.

Key words: Induced pluripotent stem cells (iPSCs); Decidua; Mesenchymal cells; X-chromosome inactivation

Received May 31, 2012; final acceptance September 30, 2012. Online prepub date: November 1, 2012.
Address correspondence to Yonehiro Kanemura, M.D., Ph.D., Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, Chuo-ku, Osaka 540-0006, Japan. Tel: +81-6-6942-1331; Fax: +81-6-6946-3530; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it


Cell Medicine, Vol. 4, pp. 149–152, 2013
2155-1790/13 $90.00 + .00
DOI: http://dx.doi.org/10.3727/215517912X653346
Copyright © 2013 Cognizant Comm. Corp.
Printed in the USA. All rights reserved

Brief Communication

Cell Persistence of Allogeneic Keratinocytes and Fibroblasts Applied in a Fibrin Matrix to Acute, Full Thickness Wounds

Jaime E. Dickerson Jr.,*† John V. Planz,‡ Barry T. Reece,§ Kathy A. Weedon,* Sandy D. Kirkpatrick,‡ and Herbert B. Slade*¶

*Healthpoint Biotherapeutics, Fort Worth, TX, USA
†Department of Cell Biology and Anatomy, University of North Texas Health Science Center, Fort Worth, TX, USA
‡Department of Forensic and Investigative Genetics, University of North Texas Health Science Center, Fort Worth, TX, USA
§RCTS, Inc., Irving, TX, USA
¶Department of Pediatrics, University of North Texas Health Science Center, Fort Worth, TX, USA

HP802-247 is a living cell suspension of cultured allogeneic growth-arrested human male keratinocytes and fibroblasts (1:9 ratio), intended for spray application to chronic wounds. In this study, a small wound was created on the arms of 28 healthy female volunteers (3-mm punch), followed by a single application of HP802-247. At each subsequent week for 8 weeks, a punch excision of the wounds was performed on a cohort of three subjects. Excised specimens were analyzed for allogeneic fibroblast and keratinocyte DNA determined by Y-chromosome short-tandem repeats using PCR amplification followed by capillary electrophoresis, a method with estimated sensitivity of 1 male cell in a background of 8,000 female cells. A complete haplotype attributable to HP802-247 fibroblasts was detected in three of three samples at 1 week, with one partial and one complete fibroblast haplotype detected at 2 weeks, and one partial keratinocyte haplotype detected at 3 weeks postapplication. The findings indicate that HP802-247 can be expected to persist in an acute wound bed for up to 2 weeks postapplication.

Key words: Allogeneic cells; Wound healing; Keratinocytes (KCs); Fibroblasts (FBs); Y-STR; HP802-247

Received February 9, 2012; final acceptance August 29, 2012. Online prepub date: October 3, 2012.
Address correspondence to Jaime E. Dickerson, Ph.D., Healthpoint Biotherapeutics, 3909 Hulen St., Fort Worth, TX 76107, USA. Tel: +1 (817) 302-3914; Fax: +1 (817) 302-3980; E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it