|Development and characterization of two new cell lines from milkfish (Chanos chanos) and grouper (Epinephelus coioides) for virus isolation|Parameswaran, V.; Ahmed, V.P.I.; Shukla, R.; Bhonde, R.R.; Hameed, A.S.S. (2007). Development and characterization of two new cell lines from milkfish (Chanos chanos) and grouper (Epinephelus coioides) for virus isolation. Mar. Biotechnol. 9(2): 281-291. dx.doi.org/10.1007/s10126-006-6110-9
In: Marine Biotechnology. Springer-Verlag: New York. ISSN 1436-2228, more
|Authors|| || Top |
- Parameswaran, V.
- Ahmed, V.P.I.
- Shukla, R.
- Bhonde, R.R.
- Hameed, A.S.S.
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°C with an optimum temperature of 28°C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 28°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 usingconfocal laser scanning microscopy (CFLSM).