Introduction of the MASH1 gene into mouse embryonic stem cells leads to differentiation of motoneuron precursors lacking Nogo receptor expression that can be applicable for transplantation to spinal cord injury.
ES cells transfected with the MASH1 gene yielded purified spinal motoneuron precursors expressing HB9 and Islet1. The cells lacked the expression of Nogo receptor that was of great advantage for axon growth after transplantation to an injured spinal cord. After transplantation, mice with the complete transection of spinal cord exhibited excellent improvement of the motor functions. Electrophysiological assessment confirmed the quantitative recovery of motor-evoked potential in the transplanted spinal cord. In the grafted spinal cord, gliosis was inhibited and Nogo receptor expression was scarcely detected. The transplanted cells labeled with GFP showed extensive outgrowth of axons positive for neurofilament middle chain, connected to each other and expressed Synaptophysin, Lim1/2 and Islet1. Thus, the in vivo differentiation into mature spinal motoneurons and the reconstitution of neuronal pathways were suggested. The grafted cell population was purified for neurons and was free from teratoma development. These therapeutic strategies may contribute to a potent treatment for spinal cord injury in future.
Department of Immunology and Medicine, St. Marianna University School of Medicine, 2-16-1, Sugao, Miyamae-ku, Kawasaki, Kanagawa 216-8511, Japan.
Mari
M
Hamada
Hideshi
H
Yoshikawa
Yuji
Y
Ueda
Manae S
MS
Kurokawa
Kenji
K
Watanabe
Manabu
M
Sakakibara
Mamoru
M
Tadokoro
Katsuya
K
Akashi
Haruhito
H
Aoki
Noboru
N
Suzuki
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D015153:Blotting, Western; D002454:Cell Differentiation; D004622:Embryo, Mammalian; D005260:Female; D005455:Fluorescent Antibody Technique; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D009046:Motor Neurons / Q000166:cytology*; D009185:Myelin Proteins / Q000378:metabolism; D009416:Nerve Regeneration / Q000502:physiology; D000070798:Nogo Proteins; D011956:Receptors, Cell Surface / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000628:therapy*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection
D051792:Basic Helix-Loop-Helix Proteins; D009185:Myelin Proteins; D000070798:Nogo Proteins; D011956:Receptors, Cell Surface; C499317:Ascl1 protein, mouse; C000603201:Rtn4 protein, mouse