title: Establishment of long term cultures of neural stem cells from adult sea bass, Dicentrarchus labrax.
abstract: 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 degrees 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.
authors.affiliation: Department of Biology, Wilfrid Laurier University, Waterloo, ON, Canada.
authors.forename: Arianna
authors.initials: A
authors.lastname: Servili
authors.forename: Mary Rose
authors.initials: MR
authors.lastname: Bufalino
authors.forename: Ryuhei
authors.initials: R
authors.lastname: Nishikawa
authors.forename: Ivan
authors.initials: I
authors.lastname: Sanchez de Melo
authors.forename: Jose A
authors.initials: JA
authors.lastname: Muñoz-Cueto
authors.forename: Lucy E J
authors.initials: LE
authors.lastname: Lee
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D001492:Bass; D001921:Brain / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation; D048430:Cell Shape; D015925:Cryopreservation; D029941:Fish Proteins / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D007700:Kinetics; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000502:physiology*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D018874:Spheroids, Cellular; D014162:Transfection
chemical_list: D029941:Fish Proteins; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; D016900:Neurofilament Proteins; D018809:Proliferating Cell Nuclear Antigen; D055748:SOXB1 Transcription Factors