Engraftable human neural stem cells

• US 6,680,198 B1
• Filed: 09/20/1999
• Issued: 01/20/2004
• Est. Priority Date: 08/14/1998
• Status: Expired due to Term

• Current Assignee: Children's Medical Center Corporation, University of British Columbia, University of Pennsylvania
• Original Assignee: Children's Medical Center Corporation, University of British Columbia, University of Pennsylvania
• Inventors: Snyder, Evan Y., Wolfe, John H., Kim, Seung U.
• Primary Examiner(s): KETTER, JAMES S
• Application Number:
  US09/398,299
• Time in Patent Office: 1,583 Days
• Field of Search: 435/368, 435/366, 435/325, 424/93.7
• US Class Current: 435/368
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0623 Stem cells

Abstract

Stable clones of neural stem cells (NSCs) have been isolated from the human fetal telencephalon. In vitro, these self-renewing clones (affirmed by retroviral insertion site) can spontaneously give rise to all 3 fundamental neural cell types (neurons, oligodendrocytes, astrocytes). Following transplantation into germinal zones of the developing newborn mouse brain, they, like their rodent counterparts, can participate in aspects of normal development, including migration along well-established migratory pathways to disseminated CNS regions, differentiation into multiple developmentally- and regionally-appropriate cell types in response to microenvironmental cues, and non-disruptive, non-tumorigenic interspersion with host progenitors and their progeny. Readily genetically engineered prior to transplantation, human NSCs are capable of expressing foreign transgenes in vivo in these disseminated locations. Further supporting their potential for gene therapeutic applications, the secretory products from these NSCs can cross-correct a prototypical genetic metabolic defect in abnormal neurons and glia in vitro as effectively as do murine NSCs. Finally, human cells appear capable of replacing specific deficient neuronal populations in a mouse model of neurodegeneration and impaired development, much as murine NSCs could. Human NSCs may be propagated by a variety of means—both epigenetic (e.g., chronic mitogen exposure) and genetic (transduction of the propagating gene vmyc)—that are comparably safe (vmyc is constitutively downregulated by normal developmental mechanisms and environmental cues) and effective in yielding engraftable, migratory clones, suggesting that investigators may choose the propagation technique that best serves the demands of a particular research or clinical problem. All clones can be cryopreserved and transplanted into multiple hosts in multiple settings.

Claims

1. A primordial human neural stem cell clone comprising both EGF and bFGF receptors maintained as a stable and identifiable cell line in vitro and suitable for on-demand implantation in vivo into a living host subject, said primordial human neural stem cell clone comprising a pluripotent and self-renewing neural stem cell of human origin, which

i) carries native human genomic DNA, which has not been genetically modified by human intervention means;

ii) remains uncommitted and undifferentiated while passaged in vitro using epigenetic means as a mitotic cell line;

iii) is capable of migrating in vivo after implantation from the implantation site to another anatomic site for in vivo integration within the nervous system of the living host subject;

iv) is capable of integrating in situ after implantation into the parenchymal tissues at a local anatomic site in the living host subject;

v) is capable of differentiating in situ into any one of neuronal cell types selected from the group consisting of neurons, oligodendroglia and astroglia as a response to signals in vivo; and

vi) expresses both EGF and bFGF receptors.

2. A living progeny of a primordial human neural stem cell clone comprising both EGF and bFGF receptors maintained as a stable and identifiable cell line in vitro and suitable for on-demand implantation in vivo into a living host subject, said primordial human neural stem cell comprising multipotent descendant cells of human neural stem cell origin which

i) carry native human genomic DNA, which has not been genetically modified by human intervention means;

ii) remain uncommitted and undifferentiated while passaged in vitro using epigenetic means as a mitotic cell line;

iii) are capable of migrating in vivo after implantation from the implantation site to another anatomic site for in vivo integration within the nervous system of the living host subject;

iv) are capable of integrating in situ after implantation into the parenchymal tissues at a local anatomic site in the living host subject;

v) are capable of differentiating in situ into any one of neuronal cell types selected from the group consisting of neurons, oligodendroglia and astroglia as a response to signals in vivo; and

vi) expresses both EGF and bFGF receptors.