title: Development and characterization of two new cell lines from milkfish (Chanos chanos) and grouper (Epinephelus coioides) for virus isolation.
abstract: Two new cell lines, SIMH and SIGE, were derived from the heart of milkfish (Chanos chanos), a euryhaline teleost, and from the eye of grouper (Epinephelus coioides), respectively. These cell lines were maintained in Leibovitz's L-15 supplemented with 20% fetal bovine serum (FBS). The SIMH cell line was subcultured more than 50 times over a period of 210 days and SIGE cell line has been subcultured 100 times over a period of 1 1/2 years. The SIMH cell line consists predominantly of fibroblastic-like cells. The SIGE cell line consists predominantly of epithelial cells. Both the cell lines were able to grow at temperatures between 25 and 32 degrees C with an optimum temperature of 28 degrees C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 28 degrees C with optimum growth at the concentrations of 15% or 20% FBS. Seven marine fish viruses were tested to determine the susceptibility of these cell lines. The SIGE cell line was found to be susceptible to nodavirus, MABV NC-1 and Y6, and the infection was confirmed by cytopathic effect (CPE) and reverse transcriptase-polymerase chain reaction. When these cells were transfected with pEGFP-N1 vector DNA, significant fluorescent signals were observed, suggesting that these cell lines can be a useful tool for transgenic and genetic manipulation studies. Further, these cell lines are characterized by immunocytochemistry using confocal laser scanning microscopy (CFLSM).
authors.affiliation: Aquaculture Biotechnology Division, Department of Zoology, C.Abdul Hakeem College, Melvisharam-632 509, Vellore Dt., Tamilnadu, India.
authors.forename: V
authors.initials: V
authors.lastname: Parameswaran
authors.forename: V P
authors.initials: VP
authors.lastname: Ishaq Ahmed
authors.forename: Ravi
authors.initials: R
authors.lastname: Shukla
authors.forename: R R
authors.initials: RR
authors.lastname: Bhonde
authors.forename: A S
authors.initials: AS
authors.lastname: Sahul Hameed
mesh_terms: D000818:Animals; D001492:Bass / Q000821:virology; D002460:Cell Line / Q000821:virology*; D002875:Chromosomes; D003470:Culture Media; D005123:Eye / Q000166:cytology*; D005399:Fishes / Q000821:virology*; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D008677:Metaphase; D009206:Myocardium / Q000166:cytology*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D013696:Temperature; D013997:Time Factors; D014779:Virus Replication
chemical_list: D003470:Culture Media