Cell transplantation of Embryonic Stem Cells for Spinal Cord Injury

2009 Fiscal Year Final Research Report

• Project/Area Number: 19591695
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cerebral neurosurgery
• Research Institution: Nara Medical University
• Principal Investigator: NAKASE Hiroyuki Nara Medical University, 医学部, 准教授 (10217739)
• Co-Investigator(Kenkyū-buntansha): NISHIMURA Fumihiko 奈良県立医科大学, 医学部, 助教 (70433331)
• Project Period (FY): 2007 – 2009
• Keywords: 脊髄損傷 / 再生治療 / 胚芽幹細胞 / 基礎研究 / マウス

Research Abstract

Embryonic stem (ES) cells are a potential source for treatment of spinal cord injury (SCI). Although one of the main problems of ES cell-based cell therapy is tumor formation, there is no ideal method to suppress tumor development. In this study, we examined whether transplantation with bone marrow stromal cells (BMSCs) prevented tumor formation in SCI model mice that received ES cell-derived grafts containing both undifferentiated ES cells and neural stem cells. Embryoid bodies (EBs) formed in 4-day hanging drop cultures were treated with retinoic acid (RA) at a low concentration of 5×10-9M for 4 days, in order to allow some of the ES cells to remain in an undifferentiated state. RA-treated EBs were enzymatically digested into single cells and used as ES cell-derived graft cells. Mice transplanted with ES cell-derived graft cells alone developed tumors at the grafted site and behavioral improvement ceased after day 21. In contrast, no tumor development was observed in mice cotransplan ted with BMSCs, which also showed sustained behavioral improvement. In vitro results demonstrated the disappearance of SSEA-1 expression in cytochemical examinations, as well as attenuated mRNA expressions of the undifferentiated markers Oct3/4, Utf1, Nanog, Sox2, and ERas by RT-PCR in RA-treated EBs cocultured with BMSCs. In addition, MAP2-immunopositive cells appeared in the EBs cocultured with BMSCs. Furthermore, the synthesis of NGF, GDNF, and BDNF was confirmed in cultured BMSCs, while immunohistochemical examinations demonstrated the survival of BMSCs and their maintained ability of neurotrophic factor production at the grafted site for up to 5 weeks after transplantation. These results suggest that BMSCs induce undifferentiated ES cells to differentiate into a neuronal lineage by neurotrophic factor production, resulting in suppression of tumor formation. Cotransplantation of BMSCs with ES cell-derived graft cells may be useful for preventing the development of ES cell-derived tumors.

Research Products

• [Journal Article] Treatment of Parkinson's disease model mice with allogeneic embryonic stem cells: necessity of immunosuppressive treatment for sustained improvement (2009)
  • Author(s): Toriumi H, Yoshikawa M, Matsuda R, Nishimura F, Yamada S, Hirabayashi H, Nakase H, Nonaka J, Ouji Y, Ishizaka S, Sakaki T
  • Journal Title: Neural Res 31 Pages: 220-227
• [Journal Article] Cotransplantation of mouse embryonic stem cells and bone marrow stromal cells following spinal cord injury suppresses tumor development (2009)
  • Author(s): Matsuda R, Yoshikawa M, Kimura H Ouji Y, Nakase H, Nishimura F, Nonaka J, Toriumi H, Yamada S Nishiofuku M, Moriya K, Ishizaka S Nakamura M, Sakaki T
  • Journal Title: Cell Transplant 18 Pages: 39-54
• [Journal Article] Protective effect of Cl esterase inhibitor on acute traumatic spinal cord injury in rat. (2008)
  • Author(s): Tei R, Nakase H, Sakaki T
  • Journal Title: Neurol Res 30(7) Pages: 761-7
• [Presentation] Co-transplantation of ES cells and bone marrow stromal cells following spinal cord in jury supPresses tumordevelopment (2007)
  • Author(s): Matsuda R, Y, shikawa M, Kimura H, Ouji Y, Nakase H, Ishizaka S, Sakaki T
  • Organizer: 2007 Shanghai International Symposium on Stem Cell Research
  • Place of Presentation: Shanghai
  • Year and Date: 2007-11-07