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Journal of Neuroscience, Vol 14, 3548-3564, Copyright © 1994 by Society for Neuroscience
Spinal cord neuroblasts proliferate in response to basic fibroblast growth factor
J Ray and FH Gage
Department of Neurosciences, University of California San Diego, La Jolla 92093-0627.
Trophic factors may function as one of the epigenic signals responsible for
the proliferation, growth, migration, and differentiation of neurons and
glia during embryogenesis. The present study reports that basic fibroblast
growth factor (bFGF) at high concentrations (10-100 ng/ml) is a mitogen for
embryonic spinal cord cells that have already committed to a neuronal
pathway and are expressing neuronal phenotypes (neuroblasts). Neuroblasts
proliferate with a doubling time of 2.5 d. To characterize the nature of
cells proliferating in response to bFGF, we have established long-term
cultures of neuroblasts that can be passaged, freeze thawed, and
recultured. In cultures the proportion of astrocytes remained the same,
indicating limited survival and proliferation of these cells in response to
bFGF. These results indicate that bFGF has mitogenic effects preferably on
neuroblasts. The morphological and biochemical characterizations of the
neuronal populations present in the long-term neuroblast cultures are
presented here. The presence of cholinergic and GABAergic neurons in the
cultures was established by immunocytochemical analysis. The cultures
contain a small number of motoneurons as judged by their immunostaining
with ChAT, low-affinity NGF receptor (LNGFR), and large size. Among all
other growth factors tested for their mitogenic effects on embryonic spinal
cord cells in culture, only epidermal growth factor (EGF) showed such
effects, but to a lesser degree. The proliferative nature of neuroblasts
has made it possible to transduce the Escherichia coli beta- galactosidase
(LacZ) gene stably into these cells in vitro using a retroviral vector. The
transfected cells expressing the foreign gene can be passaged, freeze
thawed, and recultured without the loss of transgenes. The ability to
transduce foreign genes stably into these cells permits implantation of
these cells in the spinal cord to study cellular and biochemical behaviors
and gene expression in defined neuronal populations in in vivo
environments.
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