title: Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton), and catla, Catla catla (Hamilton).
abstract: Two new cell lines, designated RE and CB, were derived from the eye of rohu, Labeo rohita, and the brain of catla, Catla catla, respectively. The cell lines were maintained in Leibovitz's L-15 supplemented with 20% foetal bovine serum. The RE cell line was sub-cultured for more than 70 passages and the CB cell line for more than 35 passages. The RE cells are rounded and consist predominantly of epithelial cells. The CB cell line consists of predominantly fibroblastic-like cells. Both cell lines are able to grow at temperatures between 25 and 32 degrees C with an optimum of 28 degrees C. The growth rate of the cells increased as the foetal bovine serum concentration increased from 2% to 20% at 28 degrees C, with optimum growth at concentrations of 15% or 20% foetal bovine serum. The cells were successfully cryopreserved and revived at different passage levels. The cell lines were not susceptible to four marine fish viruses. Extracellular products from Aeromonas sp. were toxic to the cell lines. When the cells were transfected with plasmid eukaryotic green fluorescent protein (pEGFP [Clontech, Carlsbad, CA, USA]) vector DNA, a significant fluorescent signal was observed suggesting that these cell lines could be a useful tool for transgenic and genetic manipulation studies. Polymerase chain reaction amplification of mitochondrial 12S rRNA from rohu and catla confirmed that the cell lines originated from these fish species. The cell lines were further characterized by immunocytochemistry using confocal laser scanning microscopy.
authors.affiliation: Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam, Vellore Dt., Tamilnadu, India.
authors.forename: V P Ishaq
authors.initials: VP
authors.lastname: Ahmed
authors.forename: V
authors.initials: V
authors.lastname: Chandra
authors.forename: R
authors.initials: R
authors.lastname: Sudhakaran
authors.forename: S Rajesh
authors.initials: SR
authors.lastname: Kumar
authors.forename: M
authors.initials: M
authors.lastname: Sarathi
authors.forename: V Sarath
authors.initials: VS
authors.lastname: Babu
authors.forename: B
authors.initials: B
authors.lastname: Ramesh
authors.forename: A S Sahul
authors.initials: AS
authors.lastname: Hameed
mesh_terms: D000333:Aeromonas / Q000737:chemistry; D000818:Animals; D001426:Bacterial Proteins / Q000633:toxicity; D001921:Brain / Q000166:cytology; D002460:Cell Line / Q000166:cytology / Q000187:drug effects / Q000821:virology; D015925:Cryopreservation; D003470:Culture Media / Q000737:chemistry; D003530:Cyprinidae / Q000235:genetics / Q000502:physiology* / Q000821:virology; D005123:Eye / Q000166:cytology; D050259:Genes, Mitochondrial / Q000235:genetics; D013696:Temperature; D014162:Transfection
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media