Characterization of human NT2N neurons for transplantation therapeutics / Rebecca S. Hartley.

Format:

Book

Manuscript

Microformat

Thesis/Dissertation

Author/Creator:

Hartley, Rebecca S.

Contributor:

Lee, V. M.-Y. (Virginia M.-Y.), advisor.

Nusbaum, Michael, advisor.

University of Pennsylvania.

Language:

English

Subjects (All):

Penn dissertations--Neuroscience.

Neuroscience--Penn dissertations.

Neurosciences.

Academic Dissertations as Topic.

Medical Subjects:

Neurosciences.

Academic Dissertations as Topic.

Local Subjects:

Penn dissertations--Neuroscience.

Neuroscience--Penn dissertations.

Physical Description:

vi, 129 pages : color illustrations ; 29 cm

Production:

1998.

Summary:

Stroke, Parkinson's Disease and spinal cord injury may be therapeutically treated with neuronal replacement strategies which could allow synaptically regulated delivery and circuit replacement. The NT2 cell line is the only human cell line capable of generating a pure reproducible neuronal population (NT2N neurons). I assessed the suitability of NT2N neurons for use in therapeutic transplantation. I implanted NT2N neurons into murine spinal cords and investigated their phenotypic stability and the patterning, connectivity and myelination of their process outgrowth. The effects of primary CNS cells on NT2N phenotype and the functional synaptic properties of NT2N neurons were investigated in vitro. The dopaminergic phenotype and manipulatability was examined in vitro and after CNS implantation. I found that grafted NT2N neurons fully integrate into spinal cord while maintaining a stable intrinsic phenotype. NT2N process outgrowth follows host anatomic patterns, is myelinated by the host, extends for distances $>$2 cm, and can grow within spinal nerves. Axonal morphology and synaptophysin expression suggest synaptic contacts between graft and host cells. In vitro, NT2N neurons have a manipulatable dopaminergic phenotype which is lost through mechanical disturbance. They form functional GABAergic and glutamatergic synapses. Co-culture with primary astrocytes affects NT2N morphology, survival, maturation, and synaptic function in a contact-dependent manner. I conclude that graft integration with host tissue is not dependent on neuronal progenitor cells nor on the presence of transplanted non-neuronal cells. Post-mitotic NT2N neurons are capable of phenotypic change as they differentially respond to astrocytes cultured from different CNS regions. This also indicates regional distinctions between CNS astrocytes. The lack of phenotypic effect of the host on implanted NT2N neurons appears to be due to the lack of direct contact between host and graft cells. NT2N neurons are similar to primary neurons in synaptogenesis and their survival and maturation requirements for glial support. The selective dopaminergic sensitivity to damage makes them attractive for investigations into dopaminergic phenotype and for Parkinson's Disease therapeutics. They might also be used to develop strategies for spinal cord injury treatment and to investigate the neurobiology of human neuronal transplants and for studies of neuronal development and neuron/glia interactions.

Notes:

Advisers: Virginia M.-Y. Lee; Michael Nusbaum.

Thesis (Ph.D. in Neuroscience) -- University of Pennsylvania, 1998.

Includes bibliographical references.

Local Notes:

University Microfilms order no.: 98-40198.

OCLC:

187472908