| Abstract |
Neural cell lines have been extremely useful for elucidating mechanisms of
neural cell function, regeneration and disease for mammalian species.
Despite the widely known regenerative capability of neural tissues in many
aquatic organisms, few cell lines have been derived from neural tissues of
aquatic species. In this study, we report on a newly established neural
cell line from goldfish cerebellum (GFB3C) and compare growth
characteristics in response to various parameters (temperature,
osmolality, nutritional supplements, growth/differentiation factors and
response to metals) with an aquatic invertebrate cell line (OLGA-PH-J/92)
derived from the cerebral ganglia of crayfish. Both goldfish and crayfish
have been used as model organisms to study neuronal function and survival
in polluted waters. The OLGA cell line grew best at 27 Celsius whereas
GFB3C grew well at 22-30 Celsius. For both cell lines, best growth was
observed in Leibovitz's L-15 medium supplemented with 10% Fetal Bovine
Serum, compared to other common mammalian or insect culture medias. OLGA
grew best at lower osmolalities than GFB3C cells, which was inconsistent
with their freshwater invertebrate origins. Addition of glucose (L-15
contains galactose), or growth factors did not affect growth of either
cell line. Formation of neurospheres, a characteristic of neural stem
cells was also investigated. OLGA cells did not form neurospheres, whereas
GFB3C did quite readily.
Immunostaining revealed that both cell lines consisted primarily of glial
cells as positive staining for glial fibrillary acidic protein, a glial
cell marker, was found. The cells were tested for differentiation
capabilities using retinoic acid or adhesion to poly-L-lysine as
differentiation agents. Neuronal like cellular processes could be induced
when GFB3C neurospheres were plated onto poly-L-lysine coated plates.
Finally, in terms of toxicity, both cell lines were highly tolerant to
metal exposure, which correlates with their in vivo behavior. Both could
be useful for understanding mechanisms of neural growth and
differentiation in vertebrate and invertebrate species as well as in
toxicity response and resistance in aquatic animals.
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