Abstract
We have examined primary human neuronal precursors (HNPs) from 18-22-week-old fetuses. We showed that E-NCAM/MAP2/beta-III tubulin-immunoreactive neuronal precursors divide in vitro and could be induced to differentiate into mature neurons in 2 weeks. HNPs did not express nestin and differentiated slowly compared to rodent neuronal restricted precursors (NRPs, 5 days). Immunocytochemical and physiological analyses showed that HNPs could generate a heterogeneous population of neurons that expressed neurofilament-associated protein and various neurotransmitters, neurotransmitter synthesizing enzymes, voltage-gated ion channels, and ligand-gated neurotransmitter receptors and could fire action potentials. Undifferentiated and differentiated HNPs did not coexpress glial markers. Only a subset of cells that expressed GFP under the control of the Talpha1 tubulin promoter was E-NCAM/beta-III tubulin-immunoreactive, indicating nonexclusive overlap between these two HNP cell populations. Overall, HNPs resemble NRPs isolated from rodent tissue and appear to be a neuronal precursor population.
Copyright 2001 Wiley-Liss, Inc.
Publication types
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Comparative Study
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Astrocytes / cytology
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Astrocytes / metabolism
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Bromodeoxyuridine
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Cell Differentiation / physiology
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Cells, Cultured / cytology
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Cells, Cultured / metabolism
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Central Nervous System / cytology
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Central Nervous System / embryology*
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Central Nervous System / metabolism
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Fetus / cytology
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Fetus / drug effects
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Fetus / metabolism
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Genetic Vectors
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Glial Fibrillary Acidic Protein / metabolism
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Green Fluorescent Proteins
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Humans
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Immunohistochemistry
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Intermediate Filament Proteins / metabolism
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Ion Channels / metabolism
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Luminescent Proteins / genetics
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Membrane Potentials / drug effects
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Membrane Potentials / physiology
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Nerve Tissue Proteins*
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Nestin
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Neural Cell Adhesion Molecules / metabolism
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Neurites / metabolism
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Neurites / ultrastructure
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Neurons / cytology*
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Neurons / drug effects
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Neurons / metabolism
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Neurotransmitter Agents / biosynthesis
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Neurotransmitter Agents / pharmacology
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Promoter Regions, Genetic / physiology
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Stem Cells / cytology*
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Stem Cells / drug effects
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Stem Cells / metabolism
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Tubulin / genetics
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Tubulin / metabolism
Substances
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Glial Fibrillary Acidic Protein
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Intermediate Filament Proteins
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Ion Channels
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Luminescent Proteins
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NES protein, human
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Nerve Tissue Proteins
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Nestin
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Neural Cell Adhesion Molecules
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Neurotransmitter Agents
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Tubulin
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Green Fluorescent Proteins
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Bromodeoxyuridine