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Journal of Neuroscience, Vol 12, 1802-1815, Copyright © 1992 by Society for Neuroscience
Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons
SJ Pleasure, C Page and VM Lee
David Mahoney Institute of Neurological Sciences, University of Pennsylvania School of Medicine, Philadelphia 19104-4283.
NTera 2/cl.D1 (NT2) cells, a human teratocarcinoma cell line, were
manipulated following retinoic acid treatment to yield greater than 95%
pure cultures of neuronal cells (NT2-N cells). The commitment of NT2-N
cells to a stable neuronal phenotype is irreversible as judged by the lack
of mitotic activity or phenotypic reversion over a period of 2 months in
culture. Furthermore, NT2-N cells express a variety of neuronal markers
including many neuronal cytoskeletal proteins, secretory markers, and
surface markers. NT2-N cells resemble primary neuronal cultures from
rodents morphologically and in density of process outgrowth and, like
primary neurons, go on to elaborate processes that differentiate into axons
and dendrites. This culture method yields sufficient highly differentiated
postmitotic NT2-N cells for both biochemical and molecular biological
studies. Indeed, when undifferentiated NT2 cells were stably transfected
with a beta- galactosidase (beta-gal) expression plasmid, beta-gal
expression was shown to be present in both undifferentiated NT2 and
postmitotic NT2-N cells. Thus, the ability to transfect expression plasmids
into undifferentiated NT2 cells will allow the introduction of normal and
mutant gene products into cells that can then be induced to become stable,
postmitotic human neurons. We conclude that NT2 cells and NT2-N cells
represent a unique model system for studies of human neurons, and a novel
vehicle for the expression of diverse gene products in terminally
differentiated polarized neurons.
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M. Hong, D. C. R. Chen, P. S. Klein, and V. M.-Y. Lee
Lithium Reduces Tau Phosphorylation by Inhibition of Glyc | |