Development and characterization of a new cell line from brain tissues of the rare minnow (Gobiocypris rarus)
Lv, Yuefeng | Wang, Yingying | Wang, Qing | Li, Yingying | Yin, Jiyuan | Yang, Guang | Shi, Cunbin | Li, Bo | Wang, Yahui | Bergmann, Sven M. | Zeng, Weiwei
A permanent epithelial‐like cell line, named GrB, was established from brain tissues of the rare minnow (Gobiocypris rarus). GrB cells grow well at temperatures between 22℃ and 27℃ in L‐15 medium supplemented with 10% FBS. Chromosome analysis revealed that GrB cells possessed a diploid chromosome number of 2n = 50 at passage 50, consistent with the number of chromosomes reported for G. rarus. Partial amplification and sequence analysis of the cytochrome oxidase subunit I (COI) confirmed that GrB cells are originated from G. rarus. The cell lines also successfully expressed green fluorescence protein from plasmid pEGFP‐N1 and which makes them suitable for transfection studies. A cytolytic cytopathic effect was observed in the cells after infection with tilapia lake virus 2017A (TiLV‐2017A). A large number of virus particles were detected by electron microscopy. Immunofluorescence and virus titration assays indicated that TiLV‐2017A titres reached 10⁵.⁶⁷ TCID₅₀/ml at 7 days post‐infection (dpi). Healthy adult rare minnows were challenged with virus obtained from these cells showed that they can be infected by TiLV‐2017A, which causes low mortality but clinical signs. The GrB cell line may represent a valuable model as basics for fish virology and fish immunology.
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