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Establishment of long term cultures of neural stem cells from adult sea bass, Dicentrarchus labrax

Comparative Biochemistry and Physiology, Part A 152: 245–254
Servili A., Bufalino MR, Nishikawa R, Sanchez de Melo I, Muñ,oz-Cueto JA, Lee LEJ

published: 2009 | Research publication | Recent publications

Long term cell cultures could be obtained from brains of adult sea bass (Dicentrarchus labrax) up to 5 days
post mortem. On three different occasions, sea bass brain tissues were dissected, dispersed and cultured in
Leibovitz's L-15 media supplemented with 10% fetal bovine serum. The resulting cellular preparations could be passaged within 2 or 3 weeks of growth. The neural cells derived from the first trial (SBB-W1) have now been passaged over 24 times within two years. These cells have been cryopreserved and thawed successfully. SBB-W1 cells are slow growing with doubling times requiring at least 7 days at 22 °C. These long term cell cultures could be grown in suspension as neurospheres that were immunopositive for nestin, a marker for neural stem cells, or grown as adherent monolayers displaying both glial and neural morphologies. Immunostaining with anti-glial fibrillary acidic protein ( a glial marker) and anti-neurofilament (a neuronal marker), yielded positive staining in most cells, suggesting their possible identity as neural stem cells. Furthermore, Sox 2, a marker for neural stem cells, could be detected from these cell extracts as well as proliferating cell nuclear antigen, a marker for proliferating cells. SBB-W1 could be transfected using pEGFP-N1 indicating their viability and suitability as convenient models for neurophysiological or neurotoxicological studies.

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revised Jan 4/09

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