Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production
To expand germ cell populations and provide a consistent supply for transplantation, we established basal culture conditions for sturgeon germ cells and subsequently increased their mitotic activity by eliminating gonad somatic cells, supplementing with growth factor, and replacing fetal bovine seru...
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MDPI AG
2019-03-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/9/3/106 |
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| author | Xuan Xie Ping Li Martin Pšenička Huan Ye Christoph Steinbach Chuangju Li Qiwei Wei |
| author_facet | Xuan Xie Ping Li Martin Pšenička Huan Ye Christoph Steinbach Chuangju Li Qiwei Wei |
| author_sort | Xuan Xie |
| collection | DOAJ |
| description | To expand germ cell populations and provide a consistent supply for transplantation, we established basal culture conditions for sturgeon germ cells and subsequently increased their mitotic activity by eliminating gonad somatic cells, supplementing with growth factor, and replacing fetal bovine serum (FBS). The initial basal culture conditions were Leibovitz’s L-15 medium (pH 8.0) supplemented with 5% FBS (<i>p</i> < 0.001) at 21 °C. Proliferation of germ cells was significantly enhanced and maintained for longer periods by elimination of gonad somatic cells and culture under feeder-cell free conditions, with addition of leukemia inhibitory factor and glial-cell-derived neurotrophic factor (<i>p</i> < 0.001). A serum-free culture medium improved germ cell proliferation compared to the L-15 with FBS (<i>p</i> < 0.05). Morphology remained similar to that of fresh germ cells for at least 40 d culture. Germline-specific gene expression analysis revealed no significant changes to germ cells before and after culture. Sterlet <i>Acipenser ruthenus</i> germ cells cultured more than 40 days showed development after transplant into Russian sturgeon <i>Acipenser gueldenstaedtii</i>. Polymerase chain reaction showed 33.3% of recipient gonads to contain sterlet cells after four months. This study developed optimal culture condition for sturgeon germ cells. Germ cells after 40 d culture developed in recipient gonads. This study provided useful information for culture of sturgeon germ cells. |
| first_indexed | 2024-12-10T08:18:45Z |
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| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-12-10T08:18:45Z |
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| series | Animals |
| spelling | doaj.art-3e1b14c53f33459a9ab189d1de7033402022-12-22T01:56:24ZengMDPI AGAnimals2076-26152019-03-019310610.3390/ani9030106ani9030106Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate ProductionXuan Xie0Ping Li1Martin Pšenička2Huan Ye3Christoph Steinbach4Chuangju Li5Qiwei Wei6Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, ChinaResearch Institute of Fish Culture and Hydrobiology, University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zátiší 728/II, 38925 Vodňany, Czech RepublicResearch Institute of Fish Culture and Hydrobiology, University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zátiší 728/II, 38925 Vodňany, Czech RepublicKey Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, ChinaResearch Institute of Fish Culture and Hydrobiology, University of South Bohemia in České Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zátiší 728/II, 38925 Vodňany, Czech RepublicKey Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, ChinaKey Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, ChinaTo expand germ cell populations and provide a consistent supply for transplantation, we established basal culture conditions for sturgeon germ cells and subsequently increased their mitotic activity by eliminating gonad somatic cells, supplementing with growth factor, and replacing fetal bovine serum (FBS). The initial basal culture conditions were Leibovitz’s L-15 medium (pH 8.0) supplemented with 5% FBS (<i>p</i> < 0.001) at 21 °C. Proliferation of germ cells was significantly enhanced and maintained for longer periods by elimination of gonad somatic cells and culture under feeder-cell free conditions, with addition of leukemia inhibitory factor and glial-cell-derived neurotrophic factor (<i>p</i> < 0.001). A serum-free culture medium improved germ cell proliferation compared to the L-15 with FBS (<i>p</i> < 0.05). Morphology remained similar to that of fresh germ cells for at least 40 d culture. Germline-specific gene expression analysis revealed no significant changes to germ cells before and after culture. Sterlet <i>Acipenser ruthenus</i> germ cells cultured more than 40 days showed development after transplant into Russian sturgeon <i>Acipenser gueldenstaedtii</i>. Polymerase chain reaction showed 33.3% of recipient gonads to contain sterlet cells after four months. This study developed optimal culture condition for sturgeon germ cells. Germ cells after 40 d culture developed in recipient gonads. This study provided useful information for culture of sturgeon germ cells.https://www.mdpi.com/2076-2615/9/3/106feeder cellsgerm cell cultureglial-cell-derived neurotrophic factorsturgeontransplantation |
| spellingShingle | Xuan Xie Ping Li Martin Pšenička Huan Ye Christoph Steinbach Chuangju Li Qiwei Wei Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production Animals feeder cells germ cell culture glial-cell-derived neurotrophic factor sturgeon transplantation |
| title | Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production |
| title_full | Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production |
| title_fullStr | Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production |
| title_full_unstemmed | Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production |
| title_short | Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production |
| title_sort | optimization of in vitro culture conditions of sturgeon germ cells for purpose of surrogate production |
| topic | feeder cells germ cell culture glial-cell-derived neurotrophic factor sturgeon transplantation |
| url | https://www.mdpi.com/2076-2615/9/3/106 |
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