• Journal of fish pathology
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• v.16 no.3
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• pp.165-173
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• 2003

A stable cell line, BSBS (black seabream spleen), was established from the cells in spleen of black seabream, Acanthopagrus schlegeli, and characterized. Subculture maintained more than 60 passages and mophologically, BSBS cell was epithelioid cell. The cells grew optimally at 20℃ in Leibovitz's L-15 medium supplemented with 10% fetal bovine serum with incubation temperature of 20℃. BSBS cells supported the growth of marine birnavirus (MABV Y-6), chum salmon reovirus (CSV), spring viremia of carp virus (SVCV) and hirame rhabdovirus (HIRRV). Thus, the new cell line may be useful for studying wide range of fish viruses.

• Journal of Marine Life Science
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• v.8 no.2
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• pp.186-189
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• 2023

Uncoupling protein 1 (UCP1) is a unique mitochondrial membranous protein expressed in brown adipose tissue (BAT) in mammals. While its expression in response to cold temperatures and adipogenic inducers is well-characterized in mammals and human infants, the molecular characterization and expression of UCP1 in fish remain unexplored. To address this gap, we analyzed UCP1 expression in response to adipogenic inducers in a fish cell line, rainbow trout gonadal cells (RTG-2), and compared it with UCP1 expression in three mammalian preadipocytes, 3T3-L1, T37i, and WT1 exposed to the Peroxisome proliferator-activated receptor gamma (PPARγ) agonists, rosiglitazone (Rosi). In mammalian preadipocytes, UCP1 protein was highly expressed by Rosi, with an induction of adipogenesis observed in a time-dependent manner. This suggests that UCP1 plays a significant role in adipogenesis in mammals. However, RTG-2 cells showed no response to adipogenic inducers and exhibited only marginal expressions of UCP1. These results imply that RTG-2 cells may lack crucial responsive mechanisms to adipogenic signals or that the adipogenic response is regulated by other mechanisms. Further studies are needed to confirm these phenomena in fish preadipocytes when an appropriate cell line is established in future research.

• Fisheries and Aquatic Sciences
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• v.16 no.3
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• pp.177-185
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• 2013

The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

• Journal of fish pathology
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• v.30 no.2
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• pp.71-78
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• 2017

The glycoprotein of novirhabdoviruses is known to play a critical role in the determination of host specificity. Viral hemorrhagic septicemia viruses (VHSVs) in different genotypes have different glycoprotein sequences and show different preferences for specific cell lines. In this study, to know whether the glycoprotein is solely responsible for the host cell preference of VHSV, a recombinant VHSV expressing vesicular stomatitis virus (VSV) glycoprotein instead of VHSV IVa glycoprotein (rVHSV-VSV-G) was generated by reverse genetics and inoculated into several fish cell lines, then, cytopathic effect (CPE) and viral growth caused by rVHSV-VSV-G infection were compared with those caused by rVHSV-wild that was previously generated and has the same genomic sequence with wild-type VHSV except a few nucleotides. The plaque numbers of rVHSV-VSV-G were significantly higher in EPC, BF-2 and GF cells than those of rVHSV-wild. However, in HINAE cells (originated from olive flounder), rVHSV-VSV-G titer was significantly lower than rVHSV-wild titer, and both recombinant VHSVs were not grown well in CHSE-214 cells. Although statistical significances were detected in the titers between rVHSV-wild and rVHSV-VSV-G in several cell lines, the cell line-preference order of rVHSV-VSV-G was not different from that of rVHSV-wild. These results suggest that the replacement of VHSV glycoprotein may not completely change host cell preference, and other regions of VHSV might also involve in the determination of host cell preference.

• Fisheries and Aquatic Sciences
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• v.5 no.1
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• pp.32-35
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• 2002

In this study, we investigated the mechanism of cell death in rhabdovirus-infected cells, chinook salmon embryonic cell line (CHSE-2l4) infected with hirame rhabdovirus (HIRRV). Studies using light microscopy, fluorescence microscopy, TUNEL method, electron microscopy, and agarose gel electrophoresis revealed changes in the cell morphology and DNA fragmentation indicative of apoptosis in early infection. It was observed that HIRRV induced apoptosis as well as necrosis in infected cells.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.22-23
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• 2003

Primordial germ cell (PGC) is the progenitor cell of the germ cell lineage and eventually give rise to gametes that are responsible for creating individual organisms via a fertilization process. This means that PGC is a unique cell that can be converted into individual fish. This advantage of PGCs would make it possible to develop various applications in the field of fish bioengineering. First, PGCs may make it easier to preserve the genetic resources of fish. Cryopreservation of fish eggs or embryos has not been successfully achieved so far. Therefore, the only possible method to preserve genetic resources of fishes is to raise fish as live individuals. If PGCs isolated from various fishes could be cryopresewed, these cells could be converted into live fishes via germ-line chimera production. This is particularly useful for preserving genetic materials of endangered species. Even if the species of interest were to become extinct, it could be recovered by the transplantation of cryopreserved PGCs into the embryos of a closely related species. Another application of this technology is in what could be termed "surrogate broodstock technology". (중략)

• Journal of fish pathology
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• v.5 no.1
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• pp.1-7
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• 1992

CHSE(Chinook Salmon Embryo)-214 fish cell lines was cultured in Eagle's minimal medium (MEM) supplemented with 10% fetal bovine serum and 2mM-glutamin. Optimum growth temperature of CHSE-214 cell line was $20^{\circ}C$. Infectious pancreatic necrosis virus(IPNV) was successfuly multiplied and showed the cytopathic effect in CHSE-214 cell line. Infection symptom of IPNV was observed with inverted phase contrast microscopy. At 6h-12hrs post-infection, the cells infected with IPNV were similer to normal cells. At 18-24hrs post-infection, the cells were somewhat round form and a little swollen form than normal cells. At 30hs post-infection, the cells were becoming more abnormal cells. At 48-68 post-infection, the infected cells were lysed and showed the severe cytopathic effect.

• Journal of fish pathology
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• v.34 no.1
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• pp.17-22
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• 2021

Eukaryotic translation is initiated by either cap-dependent or cap-independent way, and the cap-independent translation can be initiated by the internal ribosomal entry site (IRES). In this study, to know whether the 5'UTR leader sequence of marine birnavirus (MABV) segment A and segment B can act as IRES, bicistronic vectors harboring a CMV promoter-driven red fluorescent gene (mCherry) and poliovirus IRES- or MABV's leader sequence-driven green fluorescent gene (eGFP) were constructed, then, transfected into a mammalian cell line (BHK-21 cells) and a fish cell line (CHSE-214 cells). The results showed that the poliovirus IRES worked well in BHK-21 cells, but did not work in CHSE-214 cells. In the evaluation of MABV's leader sequences, the reporter eGFP gene under the 5'UTR leader sequence of MABV's segment A was well-translated in CHSE-214 cells, indicating 5'UTR of MABV's segment A initiates translation in the cap-independent way and can be used as a fish-specific IRES system. However, the 5'UTR leader sequence of MABV's segment B did not initiate translation in CHSE-214 cells. As the precise mechanism of birnavirid IRES-mediated translation is not known, more elaborate investigations are needed to uncover why the leader sequence of segment B could not initiate translation in the present study. In addition, further studies on the host species range of MABV's segment A IRES and on the screening of other fish-specific IRESs are needed.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.1
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• pp.65-72
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• 2020

Fish ovarian germline stem cells (OGSCs) that have the abilities to self-renew and differentiate into functional gametes can be used in various researches and applications. A main issue to be solved for effective utilization of fish OGSCs is the development of their stable in vitro culture condition, but only few researches about fish OGSC culture have been reported so far. In this study, in order to find the clues to develop the culture condition for OGSCs from Japanese medaka (Oryzias latipes), we tried to establish somatic cell lines as a candidate for the feeder cells and evaluated its supporting effects on the culture of ovarian cell populations from O. latipes. As the results, the somatic cell lines could be established only from the embryonic tissues among three tissues derived from embryos, fins and ovaries. Three embryonic cell lines were tested as a feeder cell for the culture of ovarian cell population and all three cell lines induced cell aggregation formation of the cultured ovarian cells whereas the feeder-free condition did not. Furthermore, a significant cellular proliferation was observed in the ovarian cells cultured on two of three cell lines. As a trial to increase the capacity of the cell lines as a feeder cell that supports the proliferation of the cultured ovarian cells, we subsequently established a stable line that expresses the foreign O. latipes fibroblast growth factor 2 (FGF2) from an embryonic cell line and evaluated its effectiveness as a feeder cell. The ovarian cells cultured on FGF2 expressing feeder cells still formed cell aggregates but did not show a significant increase in cellular proliferation compared to those cultured on non-transformed feeder cells. The results from this study will provide the fundamental information for in vitro culture of medaka OGSCs.

• Journal of fish pathology
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• v.16 no.2
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• pp.69-73
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• 2003

In this study, GFP reporter gene was transfected into a fibroblast cell line RTG-2 using a polycationic transfection reagent (Superfect) and showed a successful expression of GFP. The transfection efficiency by Superfect was compared to the commonly used transfection method, i.e. DNA-calcium phosphate coprecipitatlon. Transfection by Superfect was more effective than calcium phosphate coprecipltation method (frequency of cell expressing orr was 11.3% and 3.5%, respectively). The optimal expression of GFP and {\beta}-galactosidase was observed when $5-6\;{\mu}{\ell}$ of Superfect per ${\mu}g$ DNA was used for transfcction, 1:5-6 ratio between DNA(${\mu}g$) and Superfect ($\mu\ell$).