Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells
Olive flounder (<i>Paralichthys olivaceus</i>) is a commercially important fish species in Japan, China, and the Republic of Korea. Despite numerous attempts to improve productivity, there have been no studies of in vitro germline stem cell (GSC) culture in this species. Here, olive flou...
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2025-02-01
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| author | Ja Young Jo Ju-Won Kim Eun Soo Noh Yong-Ok Kim Seung Pyo Gong Hee Jeong Kong Jae Hoon Choi |
| author_facet | Ja Young Jo Ju-Won Kim Eun Soo Noh Yong-Ok Kim Seung Pyo Gong Hee Jeong Kong Jae Hoon Choi |
| author_sort | Ja Young Jo |
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| description | Olive flounder (<i>Paralichthys olivaceus</i>) is a commercially important fish species in Japan, China, and the Republic of Korea. Despite numerous attempts to improve productivity, there have been no studies of in vitro germline stem cell (GSC) culture in this species. Here, olive flounder testicular and ovarian cell lines (OFT and OFO, respectively) were established and characterized. RT-PCR demonstrated that OFT and OFO expressed several gonadal somatic cell markers, including <i>wt1</i> and <i>fgf2</i>, but lacked expression of germ cell markers, such as <i>vasa</i>, <i>nanos2</i>, and <i>scp3</i>. In addition, SNP analysis revealed that OFT originated from XY male <i>P. olivaceus</i> and OFO originated from XX female <i>P. olivaceus</i>. These results suggest that OFT was composed of Sertoli cells and OFO was composed of granulosa cells and theca cells. Finally, coculture of OFT or OFO with enriched male <i>P. olivaceus</i> GSCs isolated from the top 20% and 20–30% Percoll density gradient layers showed that GSCs were attached on both cell lines. In conclusion, we established <i>P. olivaceus</i> testicular and ovarian cell lines, which were expected to use for development of an in vitro GSC culture system. |
| format | Article |
| id | doaj-art-0beccfac2a7a4f30a362e032a4a277a2 |
| institution | DOAJ |
| issn | 2079-7737 |
| language | English |
| publishDate | 2025-02-01 |
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| spelling | doaj-art-0beccfac2a7a4f30a362e032a4a277a22025-08-20T02:42:35ZengMDPI AGBiology2079-77372025-02-0114322910.3390/biology14030229Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder CellsJa Young Jo0Ju-Won Kim1Eun Soo Noh2Yong-Ok Kim3Seung Pyo Gong4Hee Jeong Kong5Jae Hoon Choi6Biotechnology Research Division, National Institute of Fisheries Science, Busan 46083, Republic of KoreaBiotechnology Research Division, National Institute of Fisheries Science, Busan 46083, Republic of KoreaBiotechnology Research Division, National Institute of Fisheries Science, Busan 46083, Republic of KoreaBiotechnology Research Division, National Institute of Fisheries Science, Busan 46083, Republic of KoreaMajor in Aquaculture and Applied Life Science, Division of Fisheries Life Science, Pukyong National University, Busan 48513, Republic of KoreaBiotechnology Research Division, National Institute of Fisheries Science, Busan 46083, Republic of KoreaBiotechnology Research Division, National Institute of Fisheries Science, Busan 46083, Republic of KoreaOlive flounder (<i>Paralichthys olivaceus</i>) is a commercially important fish species in Japan, China, and the Republic of Korea. Despite numerous attempts to improve productivity, there have been no studies of in vitro germline stem cell (GSC) culture in this species. Here, olive flounder testicular and ovarian cell lines (OFT and OFO, respectively) were established and characterized. RT-PCR demonstrated that OFT and OFO expressed several gonadal somatic cell markers, including <i>wt1</i> and <i>fgf2</i>, but lacked expression of germ cell markers, such as <i>vasa</i>, <i>nanos2</i>, and <i>scp3</i>. In addition, SNP analysis revealed that OFT originated from XY male <i>P. olivaceus</i> and OFO originated from XX female <i>P. olivaceus</i>. These results suggest that OFT was composed of Sertoli cells and OFO was composed of granulosa cells and theca cells. Finally, coculture of OFT or OFO with enriched male <i>P. olivaceus</i> GSCs isolated from the top 20% and 20–30% Percoll density gradient layers showed that GSCs were attached on both cell lines. In conclusion, we established <i>P. olivaceus</i> testicular and ovarian cell lines, which were expected to use for development of an in vitro GSC culture system.https://www.mdpi.com/2079-7737/14/3/229testicular cellovarian cellgermline stem cellco-culture |
| spellingShingle | Ja Young Jo Ju-Won Kim Eun Soo Noh Yong-Ok Kim Seung Pyo Gong Hee Jeong Kong Jae Hoon Choi Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells Biology testicular cell ovarian cell germline stem cell co-culture |
| title | Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells |
| title_full | Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells |
| title_fullStr | Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells |
| title_full_unstemmed | Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells |
| title_short | Establishment and Characterization of OFT and OFO Cell Lines from Olive Flounder (<i>Paralichthys olivaceus</i>) for Use as Feeder Cells |
| title_sort | establishment and characterization of oft and ofo cell lines from olive flounder i paralichthys olivaceus i for use as feeder cells |
| topic | testicular cell ovarian cell germline stem cell co-culture |
| url | https://www.mdpi.com/2079-7737/14/3/229 |
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