Swaminathan T Raja, Basheer V S, Gopalakrishnan A, Sood Neeraj, Pradhan P K
National Bureau of Fish Genetic Resources Cochin Unit, CMFRI Campus, P.O. Number 1603, Cochin, 682018, Kerala, India.
Central Marine Fisheries Research Institute, P.O. Number 1603, Cochin, 682018, Kerala, India.
Cytotechnology. 2016 May;68(3):515-23. doi: 10.1007/s10616-014-9804-2. Epub 2014 Oct 31.
A new epithelial cell line, Horabagrus brachysoma fin (HBF), was established from the caudal fin tissue of yellow catfish, H. brachysoma and characterized. This HBF cell line was maintained in Leibovitz's-15 medium supplemented with 15 % fetal bovine serum (FBS) and subcultured more than 62 times over a period of 20 months. The HBF cell line consists predominantly of epithelial cells and is able to grow at temperatures between 20 and 35 °C with an optimum temperature of 28 °C. The growth rate of these cells increased as the proportion of FBS increased from 5 to 20 % at 28 °C with optimum growth at the concentrations of 15 % FBS. Partial amplification and sequencing of fragments of two mitochondrial genes 16S rRNA and COI confirmed that HBF cell line originated from yellow catfish. The HBF cells showed strong positive reaction to the cytokeratin marker, indicating that it was epithelial in nature. HBF cell line was inoculated with tissue homogenate from juveniles of Sea bass, Lates calcarifer infected with viral nervous necrosis virus (VNNV) and found not susceptible to VNNV. The extracellular products of Vibrio cholerae MTCC 3904 were toxic to the HBF cells. These cells were confirmed for the absence of Mycoplasma sp by PCR.
一种新的上皮细胞系——短吻霍氏鲶鳍细胞系(HBF),是从短吻霍氏鲶的尾鳍组织中建立并进行鉴定的。该HBF细胞系在添加15%胎牛血清(FBS)的Leibovitz's - 15培养基中培养,并在20个月内传代62次以上。HBF细胞系主要由上皮细胞组成,能够在20至35°C的温度下生长,最适温度为28°C。在28°C时,随着FBS比例从5%增加到20%,这些细胞的生长速率加快,在15% FBS浓度下生长最佳。对两个线粒体基因16S rRNA和COI的片段进行部分扩增和测序,证实HBF细胞系源自短吻霍氏鲶。HBF细胞对细胞角蛋白标记物呈强阳性反应,表明其本质上是上皮细胞。用感染病毒性神经坏死病毒(VNNV)的尖吻鲈幼鱼的组织匀浆接种HBF细胞系,发现其对VNNV不敏感。霍乱弧菌MTCC 3904的胞外产物对HBF细胞有毒性。通过PCR确认这些细胞不存在支原体。