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建立并鉴定印度鲶鱼(Clarias batrachus(L.))的尾鳍细胞系。

Establishment and characterization of a fin cell line from Indian walking catfish, Clarias batrachus (L.).

机构信息

Aquaculture Biotechnology Division, OIE Reference Laboratory for WTD, Department of Zoology, C.Abdul Hakeem College, Melvisharam, Tamilnadu, India.

出版信息

J Fish Dis. 2011 May;34(5):355-64. doi: 10.1111/j.1365-2761.2011.01247.x. Epub 2011 Mar 15.

Abstract

A new cell line, Indian Catfish Fin, derived from the fin tissue of Indian walking catfish, Clarias batrachus, was established and characterized. The cell line grew well in Leibovitz's L-15 medium supplemented with 15% foetal bovine serum (FBS) and has been subcultured more than 110 times since its initiation in 2007. The cells were able to grow at a range of temperature from 28 to 37 °C with optimal growth at 28 °C. The cell line predominantly consists of fibroblast-like cells. The growth rate of fin cells increased as the FBS concentration increased from 2% to 20% at 28 °C with optimum growth at a concentration of 15% or 20% and poor growth at a concentration of 5%. The cells were found to be susceptible to fish nodavirus and IPNV-ab and infection was confirmed by cytopathic effect and reverse transcriptase-polymerase chain reaction. PCR amplification of mitochondrial 12S rRNA using primers specific to C. batrachus confirmed the catfish origin of the cell line. The cell line was characterized further by immunocytochemistry, transfection efficiency with pEGFP-N1 and cell cycle analysis by fluorescent-activated cell sorting.

摘要

一个新的细胞系,印度鲶鱼鳍,从印度步行鲶鱼的鳍组织衍生而来,已被建立并进行了特征描述。该细胞系在添加了 15%胎牛血清(FBS)的 Leibovitz's L-15 培养基中生长良好,自 2007 年建立以来已经传代培养了 110 多次。该细胞系能够在 28 到 37°C 的温度范围内生长,最佳生长温度为 28°C。该细胞系主要由成纤维样细胞组成。在 28°C 时,随着 FBS 浓度从 2%增加到 20%,鳍细胞的生长速度增加,在浓度为 15%或 20%时生长最佳,在浓度为 5%时生长较差。通过细胞病变效应和逆转录聚合酶链反应证实,细胞对鱼神经坏死病毒和 IPNV-ab 敏感。使用针对 C. batrachus 的特异性引物对线粒体 12S rRNA 的 PCR 扩增证实了该细胞系的鲶鱼来源。通过免疫细胞化学、pEGFP-N1 的转染效率和荧光激活细胞分选的细胞周期分析进一步对该细胞系进行了特征描述。

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