Brain transplantation of immortalized human neural stem cells promotes functional recovery in mouse intracerebral hemorrhage stroke model

¹ Brain Disease Research Center, Ajou University School of Medicine, Suwon, Korea; Department of Animal Science, Korea University, Seoul, Korea; Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, Canada
² Brain Disease Research Center, Ajou University School of Medicine, Suwon, Korea; Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, Canada
³ Division of Neurology, Department of Medicine, UBC Hospital, University of British Columbia, Vancouver, Canada
⁴ National Veterinary Research and Quarantine Services, Anyang, Korea
⁵ Brain Disease Research Center, Ajou University School of Medicine, Suwon, Korea
⁶ Department of Animal Science, Korea University, Seoul, Korea
⁷ Department of Neurology, Ilsan Paik Hospital, Goyang, Korea

^* To whom correspondence should be addressed. E-mail: sukim{at}interchange.ubc.ca.

	Abstract

We have generated stable, immortalized cell lines of human neural stem cells (NSC) from primary human fetal telencephalon cultures via a retroviral vector encoding v-myc. HB1.F3, one of the human NSC lines, express normal human karyotype of 46,XX and nestin, a cell type-specific marker for NSC. F3 has the ability to proliferate continuously, and differentiate into cells of neuronal and glial lineage. HB1.F3 human NSC line was used for cell therapy in mouse model of intracerebral hemorrhage (ICH) stroke. Experimental ICH was induced in adult mice by intrastriatal administration of bacterial collagenase; one week after surgery, the mice were randomly divided into 3 groups so as to receive intracerebrally either F3 human NSCs labeled with -galactosidase (n=31), killed F3 NSCs (n= 21) or PBS (n=30). Transplanted NSCs were detected by X-gal histochemistry or double labeling with -gal and MAP2, neurofilaments (both for neurons) or GFAP (for astrocytes). Behavior of the animals was evaluated for period up to 8 weeks using modified rotarod tests and limb placement test. Transplanted human NSCs were identified in the peri-hematomal areas and differentiated into neurons (-gal/ MAP2+ and -gal/ NF+) or astrocytes (-gal / GFAP+). The NSC transplanted group showed markedly improved functional performance on rotarod test and limb placement after 2-8 weeks as compared with the control killed F3 NSC (p,0.001) or PBS groups (p<0.001). These results indicate that the stable immortalized human NSCs are a valuable source of cells for cell replacement and gene transfer for the treatment of ICH and other human neurological disorders.