Cell transplantation of Embryonic Stem Cells for Spinal Cord Injury

• 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
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2009: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000); Fiscal Year 2008: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 脊髄損傷 / 再生治療 / 胚芽幹細胞 / 基礎研究 / マウス / 補体活性抑制 / 脊髄血流

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] 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
  • Peer Reviewed
• [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
• [Journal Article] Protective effect of Cl esterase inhibitor on acute traumatic spinal cord injury in the rat. (2008)
  • Author(s): Tei R., Kaido T., Nakase H., Sakaki T.
  • Journal Title: Neuro Res 30 Pages: 761-767
  • Peer Reviewed
• [Journal Article] Protective effect of Cl esterase inhibitor on acute traumatic spinal cord injury in the rat.
  • Author(s): Tei R
  • Journal Title: Neurol Res. (in press)
  • Peer Reviewed
• [Presentation] 頚椎・頚髄損傷に対する外科的治療. ～当科での方針と治療成績～ (2009)
  • Author(s): 中瀬裕之, 竹島靖浩, 本山靖, 朴永銖, 平林秀裕, 榊寿右
  • Organizer: 第32回日本神経外傷学会
  • Place of Presentation: 下関
  • Year and Date: 2009-04-18
• [Presentation] 脊髄損傷に対するES細胞移植治療の問題点 (2008)
  • Author(s): 松田良介, 中瀬裕之, 木村新, 西村文彦, 吉川正英, 榊寿右
  • Organizer: 第23回日本脊髄外科学会
  • Place of Presentation: 宮城
  • Year and Date: 2008-06-12
• [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
• [Presentation] Co-transplalltation of ES cells and bone marrow stromal cells following spinal cord injury suppresses tumor develoment. (2007)
  • Author(s): Matsuda R
  • Organizer: 2007 Shanghai International Symposium on Stem Cell Research
  • Place of Presentation: Shanghai
  • Year and Date: 2007-11-07