Establishment of embryonic cell line from sea bass (Lates calcarifer) for virus isolation.
A continuous cell line was established from blastula stage embryos of sea bass (Lates calcarifer). The sea bass embryonic cells were maintained in Leibovitz's L-15 supplemented with 15% fetal bovine serum. The embryonic cell line was sub-cultured more than 70 passages over a period of 1.5 years and is designated as Sahul Indian sea bass embryonic (SISE) cell line. The cells were able to grow at temperatures between 25 and 32 degrees C with an optimum temperature of 28 degrees C. The growth rate of sea bass embryonic cells increased as the FBS proportion increased from 2 to 20% at 28 degrees C with optimum growth at the concentration of 15 or 20%. Polymerase chain reaction products were obtained from embryonic cells and blastula of sea bass with primer sets of microsatellite markers of sea bass. Four fish viruses were tested on this cell line to determine its susceptibility to these viruses and this cell line was found to be susceptible to IPNV VR-299 and nodavirus, and the infection was confirmed by cytopathic effect (CPE) and RT-PCR. Further, this cell line was characterized by immunocytochemistry using confocal-laser-scanning microscopy (CFLSM), transfection with pEGFP-N1, proliferate marker (BrdU).
Parameswaran
V
V
Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam 632509, Vellore District, Tamilnadu, India.
Shukla
Ravi
R
Bhonde
Ramesh
R
Hameed
A S Sahul
AS
D000818:Animals; D001492:Bass; D002417:Cattle; D002460:Cell Line; D003470:Culture Media; D003588:Cytopathogenic Effect, Viral; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D029142:Infectious hematopoietic necrosis virus / Q000235:genetics / Q000254:growth & development*; D018061:Infectious pancreatic necrosis virus / Q000235:genetics / Q000254:growth & development*; D008853:Microscopy; D046529:Microscopy, Electron, Transmission; D030381:Nodaviridae / Q000235:genetics / Q000254:growth & development*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum; D013696:Temperature; D014773:Virology / Q000379:methods*
D003470:Culture Media; D012367:RNA, Viral; D049452:Green Fluorescent Proteins