Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions
A primary cell culture derived from the gill tissues of largemouth bass (<i>Micropterus salmoides</i>) was successfully established and characterized, providing a physiologically relevant model for virological research. Gill tissues were enzymatically dissociated, and their cells were cu...
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2025-01-01
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| Online Access: | https://www.mdpi.com/2410-3888/10/1/18 |
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| author | Ziwen Wang Li Nie Chenjie Fei Jiong Chen |
| author_facet | Ziwen Wang Li Nie Chenjie Fei Jiong Chen |
| author_sort | Ziwen Wang |
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| description | A primary cell culture derived from the gill tissues of largemouth bass (<i>Micropterus salmoides</i>) was successfully established and characterized, providing a physiologically relevant model for virological research. Gill tissues were enzymatically dissociated, and their cells were cultured in M199 supplemented with 20% fetal bovine serum at 25 °C, yielding optimal growth. Viral replication within these primary cells was confirmed by transmission electron microscopy, and further qRT-PCR demonstrated the upregulation of antiviral genes (<i>IFN1</i>, <i>Mx1</i>, <i>ISG15</i>, and <i>Viperin</i>). These primary gill cells of spindle-like morphology exhibited significantly higher susceptibility to <i>Micropterus salmoides</i> rhabdovirus (MSRV) compared to established cell lines, as evidenced by higher viral titers, thus establishing their suitability for studying host–virus interactions. Furthermore, these cells were amenable to genetic manipulation, with the successful transfection of an mCherry reporter gene using commercially available reagents. These findings highlight the utility of the largemouth bass gill-derived primary cell culture as an alternative in vitro system for investigating MSRV pathogenesis and host immune responses, which serves as a stepping stone for improved antiviral strategies in largemouth bass aquaculture. |
| format | Article |
| id | doaj-art-5332cf03efa74457af4f273c821877eb |
| institution | Kabale University |
| issn | 2410-3888 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Fishes |
| spelling | doaj-art-5332cf03efa74457af4f273c821877eb2025-01-24T13:32:27ZengMDPI AGFishes2410-38882025-01-011011810.3390/fishes10010018Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus InteractionsZiwen Wang0Li Nie1Chenjie Fei2Jiong Chen3State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, School of Marine Sciences, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, School of Marine Sciences, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, School of Marine Sciences, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, School of Marine Sciences, Ningbo University, Ningbo 315211, ChinaA primary cell culture derived from the gill tissues of largemouth bass (<i>Micropterus salmoides</i>) was successfully established and characterized, providing a physiologically relevant model for virological research. Gill tissues were enzymatically dissociated, and their cells were cultured in M199 supplemented with 20% fetal bovine serum at 25 °C, yielding optimal growth. Viral replication within these primary cells was confirmed by transmission electron microscopy, and further qRT-PCR demonstrated the upregulation of antiviral genes (<i>IFN1</i>, <i>Mx1</i>, <i>ISG15</i>, and <i>Viperin</i>). These primary gill cells of spindle-like morphology exhibited significantly higher susceptibility to <i>Micropterus salmoides</i> rhabdovirus (MSRV) compared to established cell lines, as evidenced by higher viral titers, thus establishing their suitability for studying host–virus interactions. Furthermore, these cells were amenable to genetic manipulation, with the successful transfection of an mCherry reporter gene using commercially available reagents. These findings highlight the utility of the largemouth bass gill-derived primary cell culture as an alternative in vitro system for investigating MSRV pathogenesis and host immune responses, which serves as a stepping stone for improved antiviral strategies in largemouth bass aquaculture.https://www.mdpi.com/2410-3888/10/1/18primary cell cultureslargemouth bass<i>Micropterus salmoides</i> rhabdovirus |
| spellingShingle | Ziwen Wang Li Nie Chenjie Fei Jiong Chen Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions Fishes primary cell cultures largemouth bass <i>Micropterus salmoides</i> rhabdovirus |
| title | Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions |
| title_full | Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions |
| title_fullStr | Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions |
| title_full_unstemmed | Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions |
| title_short | Establishment of Gill-Derived Primary Cell Cultures from Largemouth Bass (<i>Micropterus salmoides</i>) as an Alternative Platform for Studying Host–Virus Interactions |
| title_sort | establishment of gill derived primary cell cultures from largemouth bass i micropterus salmoides i as an alternative platform for studying host virus interactions |
| topic | primary cell cultures largemouth bass <i>Micropterus salmoides</i> rhabdovirus |
| url | https://www.mdpi.com/2410-3888/10/1/18 |
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