Cell transplantation for the treatment of spinal cord injury in rats-morphological study and search for humoral effective factors-

2010 Fiscal Year Final Research Report

• Project/Area Number: 20300122
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Aino University
• Principal Investigator: IDE Chizuka Aino University, 医療保健学部, 教授 (70010080)
• Co-Investigator(Kenkyū-buntansha): NAKANO Norihiko 藍野大学, 療保健学部, 准教授 (40322721) ; YAMADA Yoshihiro 藍野大学, 療保健学部, 教授 (30252464)
• Co-Investigator(Renkei-kenkyūsha): SUZUKI Yoshihisa 田附興風会, 野病院, 形成外科部長 (30243025) ; DEZAWA Mari 京都大学, 医学研究科, 准教授 (50272323) ; TAGUCHI Naoyuki 大阪大学, 微生物研究所, 教授 (90002188)
• Project Period (FY): 2008 – 2010
• Keywords: 神経再生 / 移植・再生医療 / 脊髄再生 / 再生医学 / 栄養因子 / 骨髄間質細胞

Research Abstract

(1) Transplantation of bone marrow stromal cells (BMSCs)
(a) Direct injection of BMSCs into the spinal cord lesion BMSCs (5×10^5) were injected into the spinal cord lesion 2 weeks after injury, and rats were observed for 4 weeks post-transplantation. Locomotion of rats was recovered up to about 10 points of BBB scale, with the control points of 4-5. Numerous regenerating axons extended through the lesion from the rostral to distal border. Electron microscopy showed that regenerating axons were surrounded by Schwann cells in the collagen fibril matrices. BMSCs disappeared within 2 weeks post transplantation, indicating that they were never incorporated into the spinal cord.
(b) Transplantation through the cerebrospinal fluid (CSF)
BMSCs (5×10^5) were injected into the 4^<th> ventricle 1, 2, and 4 weeks after spinal cord injury. Locomotion of cell-injected rats was recovered up to 10-13 points of BBB scale, while those of control remained at 0-4. Regenerating axons extended in bundles bridg ing the lesion between the rostral and distal margin. Regenerating axons extended through the collagen fibril matrices and were surrounded by Schwann cells. This shows that regenerating axons are in the same environment as peripheral nerves. Injected BMSCs disappeared from the spinal cord 2 weeks after injection, indicating that BMSCs exert their effects not by integration into the spinal cord, but probably by secreting some trophic factors in to the CSF.
(2) Effective factors contained in the conditioned medium of BMSCs It was demonstrated that the conditioned medium (CM) contained IGF-1, HGF, VEGF, and TGF-β1. Cultured hippocampal neurons had receptors of IGF-1 and TGF-β1. They showed no changes in density by the treatment of CM. These trophic factors, when they were used single or in combination of 2, 3 and 4 factors to treat cultured hippocampal neurons in vitro, did not show the same effect level of cell viability and neurite extension as the case of CM. This suggests that the CM may contain some unidentified trophic factors.

Research Products

• [Journal Article] Characterization of conditioned medium of cultured bone marrow stromal cells. (2010)
  • Author(s): Nakano N, Nakai Y, Seo TB, Yamada Y, Ohno O, Yamanaka A, Nagai Y, Fukushima M, Suzuki Y, Nakatani N, Ide C
  • Journal Title: Neurosci Lett 483 Pages: 57-61
• [Journal Article] Bone Marrow Stromal Cell Transplantation for Treatment of Sub-acute Spinal Cord Injury in the Rat. (2010)
  • Author(s): Ide C, Nakai Y, Nakano N, Seo TB, Yamada Y, Endo K, Noda T, Saito F, Suzuki Y, Fukushima M, Nakatani T
  • Journal Title: Brain Res 1332 Pages: 32-47
• [Journal Article] Transplantation of cultured choroid plexus epithelial cells via cerebrospinal fluid shows prominent neuroprotective effects against acute ischemic brain injury in the rat. (2010)
  • Author(s): Matsumoto N, Taguchi A, Kitayama H, Watanabe Y, Ohta M, Yoshihara T, Itokazu Y, Dezawa M, Suzuki Y, Sugimoto H, Noda M, Ide C
  • Journal Title: Neurosci Lett 469 Pages: 283-288
• [Journal Article] Locomotor improvement after autologous bone marrow stromal cell intrathecal delivery in dogs with chronic spinal cord injury.
  • Author(s): Nishida H, Nakayama M, Tanaka H, Kitamura M, Hatoya S, Sugiura K, Suzuki Y, Ide C, Inaba T
  • Journal Title: American Journal of Veterinary Research (in press)
• [Journal Article] Bone marrow stromal cell transplantation in spinal cord injury.
  • Author(s): Nakai Y, Nakano N, Seo TB, Yamada Y, Noda T, Ide C
  • Journal Title: in Bone Marrow Transplantation : Indications, Procedures and Outcomes(Nova Science Publishes, NY, USA). (in press)