Establishment of a new fish cell line from the brain of humpback grouper (Cromileptes altivelis) and its application in toxicology and bacterial susceptibility

Liu, Yixuan; Wei, Caoying; Liu, Zhiru; Cao, Zhenjie; Sun, Yun; Zhou, Yongcan; Wang, Shifeng; Guo, Wei

doi:10.1007/s10695-021-01006-8

Cited by 10 publications

(4 citation statements)

References 53 publications

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“…18S rRNA has frequently been used in species identification and classification among aquatic and mammalian species (Liu et al . 2021 ; Zheng and Yang 2020 ). The sequencing of 18S rRNA indicated that the SMI cells were derived from turbot.…”

Section: Discussionmentioning

confidence: 99%

Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Liu

et al. 2023

In Vitro Cell.Dev.Biol.-Animal

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A continuous intestine cell line from turbot (Scophthalmus maximus) designated as SMI was established utilizing the tissue explant technique. Primary SMI cell was cultured at 24 °C in a medium with 20% fetal bovine serum (FBS), then subcultured in 10% FBS after 10 passages. Impacts of medium or temperature on the growth of SMI were examined and the results indicated it grew well in DMEM supplemented with 10% FBS at 24 °C. The SMI cell line was subcultured more than 60 times. Karyotyping, chromosome number, and ribosomal RNA genotyping analysis revealed that SMI had a modal diploid chromosome number of 44 and originated from turbot. After being transfected with pEGFP-N1 and FAM-siRNA, a large number of green fluorescence signals were observed in SMI, indicating that SMI could be used as an ideal platform to explore gene function in vitro. In addition, the expression of epithelium-associated genes such as itga6, itgb4, gja1, claudin1, zo-1, and E-cadherin in SMI suggested the SMI had some characteristics of epidermal cells. The upregulation of immune-associated genes such as TNF-β, NF-κB, and IL-1β in SMI after stimulation with pathogen-associated molecular patterns suggested the SMI might exhibit immune functions similar to the intestinal epithelium in vivo.

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Section: Discussionmentioning

confidence: 99%

Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Liu

et al. 2023

In Vitro Cell.Dev.Biol.-Animal

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“…Some fish brain cell lines have been established for NNV pathogenesis studies, such as Lates calcarifer and Lateolabrax japonicus [ 51 , 52 ]. According to our previous study, the CAB cell line is susceptive to Vibrio harveyi and Edwardsiella tarda [ 21 ]. In the current study, we observed typical CPEs after NNV-infection, and the virus replication was also confirmed by qRT-PCR.…”

Section: Discussionmentioning

confidence: 99%

“…The CAB ( C. altivelis brain) cell line was established by our lab and cultured in Leibovitz’s L-15 medium (Gibco, Grand Island, NY, USA) supplemented with 15% fetal bovine serum (FBS, Gibco) at 26 °C [ 21 ]. The viral stock was titrated in CAB cells by 50% tissue culture infectious dose (TCID 50 ) determined as described previously [ 22 ].…”

Section: Methodsmentioning

confidence: 99%

Cromileptes altivelis microRNA Transcriptome Analysis upon Nervous Necrosis Virus (NNV) Infection and the Effect of cal-miR-155 on Cells Apoptosis and Virus Replication

Cao

Liu

et al. 2022

Viruses

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MicroRNAs (miRNAs) could regulate various biological processes. Nervous necrosis virus (NNV) is one of the primary germs of the Humpback grouper (Cromileptes altivelis), a commercial fish of great importance for Asian aquaculture. However, there is limited available information on the host-virus interactions of C. altivelis. miRNAs have been shown to play key roles in the host response to infection by a variety of pathogens. To better understand the regulatory mechanism of miRNAs, we constructed miRNA transcriptomes and identified immune-related miRNAs of C. altivelis spleen in response to NNV infection. Reads from the three libraries were mapped onto the Danio rerio reference genome. As a result, a total of 942 mature miRNAs were determined, with 266 known miRNAs and 676 novel miRNAs. Among them, thirty-two differentially expressed miRNAs (DEmiRs) were identified compared to the PBS control. These DEmiRs were targeted on 895 genes, respectively, by using miRanda v3.3a. Then, 14 DEmiRs were validated by qRT-PCR and showed consistency with those obtained from high-throughput sequencing. In order to study the relationship between viral infection and host miRNA, a cell line from C. altivelis brain (CAB) was used to examine the expressions of five known DEmiRs (miR-132-3p, miR-194a, miR-155, miR-203b-5p, and miR-146) during NNV infection. The results showed that one miRNA, cal-miRNA-155, displayed significantly increased expression in response to the virus infection. Subsequently, it was proved that overexpression of cal-miR-155 enhanced cell apoptosis with or without NNV infection and inhibited virus replication in CAB cells. Oppositely, the cal-miRNA-155 inhibitor markedly suppressed apoptosis in CAB cells. The results of the apoptosis-related genes mRNA expression also showed the regulation of cal-miR-155 on the apoptosis process in CAB cells. These findings verify that miR-155 might exert a function as a pro-apoptotic factor in reply to NNV stimulation in CAB cells and help us further study the molecular mechanisms of the pathogenesis of NNV in C. altivelis.

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“…To date, 783 cell lines have been developed from marine and fresh water fish species (Thangaraj et al., 2021). Fish cell line is a potential in vitro tool which deals with a variety of applications including screening of viruses followed by viral propagation, vaccine development, (Suryakodi et al., 2021; Zeng et al., 2022), toxicology (Ashafque et al., 2022; Liu et al., 2021), immunological gene expression (Rakus et al., 2019; Ruiz‐Palacios et al., 2020) and host–pathogen interaction (Faber et al., 2021; Wang et al., 2020). The fish cell lines are widely applied in virological and toxicological studies.…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of brain cell line from Trachinotus blochii and its application in virological and gene expression studies

Nafeez Ahmed,

Mithra,

Suryakodi

et al. 2024

Journal of Fish Diseases

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A permanent cell line, SPB (Snubnose pompano brain) was established from Trachinotus blochii by the explant culture method. It has been sub‐cultured more than 75 passages and showed optimal growth at 28°C using L‐15 medium supplemented with 15% to 20% FBS. The SPB cells were cryopreserved at different passage levels for various applications. SPB cells were composed of fibroblastic and epithelial‐like cells. The SPB cells were tested for mycoplasma contamination which was found to be negative. The origin of the SPB cell line from T. blochii was confirmed by amplification of the mitochondrial cytochrome oxidase I (COI) gene. The transfection efficiency of SPB cell line is 15% assessed by expression of green fluorescent protein using pEGFP‐N1 plasmid. In addition, two CMV promotor plasmids pFNCPE42‐DNA and pcDNAVP28 were transfected to SPB cells and it shows high expression levels of FNCP of fish nodavirus and VP28 protein of white spot syndrome virus by immunostaining. The SPB cells showed susceptibility to SJNNV and the infection was confirmed by RT‐PCR, Western blot, ELISA, TCID50 and RT‐qPCR. Experimental infection was carried out in T. blochii using SJNNV propagated in SPB cell line and found 100% mortality with clinical signs. The infection was confirmed by RT‐PCR. The SPB cell line can be used for propagation of fish viral pathogens and production of the recombinant proteins.

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Establishment of a new fish cell line from the brain of humpback grouper (Cromileptes altivelis) and its application in toxicology and bacterial susceptibility

Cited by 10 publications

References 53 publications

Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Cromileptes altivelis microRNA Transcriptome Analysis upon Nervous Necrosis Virus (NNV) Infection and the Effect of cal-miR-155 on Cells Apoptosis and Virus Replication

Development and characterization of brain cell line from Trachinotus blochii and its application in virological and gene expression studies

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