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Lucy Lee

Characterization of OLGA PH J/92, a cell line derived from neural tissue of the freshwater crayfish

and comparison with a goldfish brain cell line
MaryRose Bufalino

published: 2008 | Thesis | Honours BSc

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 ?C whereas GFB3C grew well at 22-30 ?C. 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.

**** NOTE:SUBSEQUENT MOLECULAR EVALUATION OF THE OLGA CELL LINE PROVED NEGATIVE FOR ORCONECTES LIMOSUS, WHICH WAS ALSO CONFIRMED BY ATCC, THUS OLGA IS NOT A CRUSTACEAN CELL LINE ****

Download: PDF (6.33MB) Bufalino_BSc_Thesis.pdf

revised Jun 30/08

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