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Journal of Neuroscience, Vol 6, 3070-3078, Copyright © 1986 by Society for Neuroscience
Schwann cells stimulated to proliferate in the absence of neurons retain full functional capability
S Porter, MB Clark, L Glaser and RP Bunge
Schwann cells from neonatal rat sciatic nerve can be maintained and grown
in culture in the absence of neurons. We are interested in substantially
expanding such cultures for use in the study of Schwann cells, their growth
responses, and their interactions with neurons. However, it was important
to determine if expanded cell populations retained their distinguishing
biological properties and their ability to differentiate when recombined
with neurons. Therefore, we have compared the functional properties of
extensively expanded populations of sciatic nerve Schwann cells to those of
embryonic dorsal root ganglion (DRG) Schwann cells that had been briefly
expanded in vitro in the continuous presence of ganglion neurons. Sciatic
nerve Schwann cells were cultured and purified according to the methods of
Brockes et al. (1979). A combination of crude glial growth factor and
forskolin was found to act synergistically in providing maximal stimulation
of Schwann cell proliferation. Sciatic nerve Schwann cells that were
continuously expanded for at least 2 months were compared to Schwann cells
derived from fetal dorsal root ganglia. The results indicate that the
complement of secreted proteins from both cell populations, either in
isolation or recombined with neurons, was essentially identical; both cell
populations expressed the cell-surface antigens laminin and Ran 1 (217C
antibody); after seeding onto DRG neurons, both cell populations associated
with neuronal processes with the same time course; and under identical
nutrient conditions, both cell populations were observed to exhibit a
comparable capacity for myelination of DRG axons in vitro. Thus, methods
used to establish primary cultures of rat sciatic nerve Schwann cells and
to expand secondary cultures in vitro in the absence of neurons preserve
basic Schwann cell functions.
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