Oligodendrocytes and Radial Glia Derived From Adult Rat Spinal Cord Progenitors: Morphological and Immunocytochemical Characterization

Iris Kulbatski ^1*, Andrea J. Mothe ¹, Armand Keating ¹, Yoji Hakamata ¹, Eiji Kobayashi ¹ and Charles H. Tator ¹

¹ Institute of Medical Science, University of Toronto, Toronto, Ontario, Canada (IK,AK,CHT); Division of Cellular and Molecular Biology, Toronto Western Research Institute, University Health Network, Toronto, Ontario, Canada (IK,AJM,CHT); Princess Margaret Hospital/Ontario Cancer Institute, Toronto, Ontario, Canada (AK); and Divisions of Organ Replacement Research and Animal Resource Project, Centre for Molecular Medicine, Jichi Medical School, Kawachi, Tochigi, Japan (YH,EK)

^* To whom correspondence should be addressed. E-mail: iris.kulbatski{at}sympatico.ca.

Submitted on May 18, 2006
Accepted on 25 October 2006

	Abstract

Self-renewing, multipotent neural progenitor cells (NPCs) reside in the adult mammalian spinal cord ependymal region. The current study characterized, in vitro, the native differentiation potential of spinal cord NPCs isolated from adult enhanced green fluorescence protein rats. Neurospheres were differentiated, immunocytochemistry (ICC) was performed, and the positive cells were counted as a percentage of Hoescht+ nuclei in 10 random fields. Oligodendrocytes constituted most of the NPC progeny (58.0% of differentiated cells; 23.4% in undifferentiated spheres). ICC and electron microscopy (EM) showed intense myelin production by neurospheres and progeny. The number of differentiated astrocytes was 18.0%, but only 2.8% in undifferentiated spheres. The number of differentiated neurons was 7.4%, but only 0.85% in undifferentiated spheres. The number of differentiated radial glia (RG) was 73.0% and in undifferentiated spheres 80.9%. EM showed an in vitro phagocytic capability of NPCs. The number of undifferentiated NPCs was 32.8% under differentiation conditions and 78.9% in undifferentiated spheres. Compared to ependymal region spheres, the spheres derived from the peripheral white matter of the spinal cord produced glial restricted precursors. These findings indicate that adult rat spinal cord ependymal NPCs differentiate preferentially into oligodendrocytes and RG, which may support axonal regeneration in future trials of transplant therapy for spinal cord injury.

Key Words: spinal cord injury, neural progenitors, regeneration, differentiation, phenotypic characterization, oligodendrocyte, radial glia

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