Parameswaran V, Shukla Ravi, Bhonde Ramesh, Hameed A S Sahul
Aquaculture Biotechnology Division, Department of Zoology, C. Abdul Hakeem College, Melvisharam 632509, Vellore District, Tamilnadu, India.
J Virol Methods. 2006 Nov;137(2):309-16. doi: 10.1016/j.jviromet.2006.07.006. Epub 2006 Aug 21.
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).
从尖吻鲈（Lates calcarifer）囊胚期胚胎建立了一个连续细胞系。尖吻鲈胚胎细胞在添加15%胎牛血清的Leibovitz's L - 15培养基中培养。该胚胎细胞系在1.5年的时间内传代培养超过70次，被命名为萨胡尔印度尖吻鲈胚胎（SISE）细胞系。细胞能够在25至32摄氏度的温度下生长，最适温度为28摄氏度。在28摄氏度时，随着胎牛血清比例从2%增加到20%，尖吻鲈胚胎细胞的生长速率加快，在15%或20%的浓度下生长最佳。用尖吻鲈微卫星标记引物组从尖吻鲈胚胎细胞和囊胚中获得了聚合酶链反应产物。在该细胞系上测试了四种鱼类病毒以确定其对这些病毒的敏感性，发现该细胞系对传染性胰腺坏死病毒VR - 299和诺达病毒敏感，并通过细胞病变效应（CPE）和逆转录 - 聚合酶链反应（RT - PCR）证实了感染。此外，使用共聚焦激光扫描显微镜（CFLSM）通过免疫细胞化学、用pEGFP - N1转染、增殖标记物（BrdU）对该细胞系进行了表征。
