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Engraftable human neural stem cells respond to development cues, replace neurons, and express foreign genes

Abstract

Stable clones of neural stem cells (NSCs) have been isolated from the human fetal telencephalon. These self-renewing clones give rise to all fundamental neural lineages in vitro. Following transplantation into germinal zones of the newborn mouse brain they participate in aspects of normal development, including migration along established migratory pathways to disseminated central nervous system regions, differentiation into multiple developmentally and regionally appropriate cell types, and nondisruptive interspersion with host progenitors and their progeny. These human NSCs can be genetically engineered and are capable of expressing foreign transgenes in vivo. Supporting their gene therapy potential, secretory products from NSCs can correct a prototypical genetic metabolic defect in neurons and glia in vitro. The human NSCs can also replace specific deficient neuronal populations. Cryopreservable human NSCs may be propagated by both epigenetic and genetic means that are comparably safe and effective. By analogy to rodent NSCs, these observations may allow the development of NSC transplantation for a range of disorders.

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Figure 1: Southern blot analysis of retroviral insertion into human NSC clones.
Figure 2: Characterization of human NSCs in vitro.
Figure 3: Dissociated brain cells from mice with mutated α-subunit of β-hexosaminidase (Tay-Sachs disease) cocultured with human NSCs.
Figure 4: Migration of human NSCs following engraftment into the SVZ of newborn mice.
Figure 5: Characterization of human NSC clones in vivo following engraftment into SVZ of neonatal mice.
Figure 6: Transplantation of human NSCs into granule neuron-deficient cerebellum.

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Acknowledgements

We thank Angelo Vescovi, Ron McKay, and Jeff Macklis for advice and critique, and Baolin Chang for technical assistance. This work was supported in part by grants to E.Y.S. from NINDS (NS33852), the Paralyzed Veterans of America, the American Paralysis Association, Late Onset Tay-Sachs Foundation, and Hood Foundation; to J.H.W. from NIDDK (DK42707; DK46637); to S.U.K. from the Myelin Project of Canada; and to R.L.S. from NINDS (NS20820). J.D.F. and S.A. were partially supported by NIH training grants.

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Departments of Neurology, Pediatrics, and Neurosurgery, Children’s Hospital, Harvard Medical School, Boston, MA.

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Flax, J., Aurora, S., Yang, C. et al. Engraftable human neural stem cells respond to development cues, replace neurons, and express foreign genes. Nat Biotechnol 16, 1033–1039 (1998). https://doi.org/10.1038/3473

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