Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines.
Primordial germ cells (PGCs) are precursors of functional gametes and can be used as efficient transgenic tools and carriers in bioreactors. Few methods for long-term culture of PGCs are available. In this study, we tested various culture conditions for PGCs, and used the optimum culture system to c...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2018-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5927411?pdf=render |
| _version_ | 1811319923559890944 |
|---|---|
| author | Linglin Kong Lingling Qiu Qixin Guo Ying Chen Xin Zhang Bowen Chen Yang Zhang Guobin Chang |
| author_facet | Linglin Kong Lingling Qiu Qixin Guo Ying Chen Xin Zhang Bowen Chen Yang Zhang Guobin Chang |
| author_sort | Linglin Kong |
| collection | DOAJ |
| description | Primordial germ cells (PGCs) are precursors of functional gametes and can be used as efficient transgenic tools and carriers in bioreactors. Few methods for long-term culture of PGCs are available. In this study, we tested various culture conditions for PGCs, and used the optimum culture system to culture chicken gonad PGCs for about three hundred days. Long-term-cultured PGCs were detected and characterized by karyotype analysis, immunocytochemical staining of SSEA-1, c-kit, Sox2, cDAZL, and quantitative RT-PCR for specific genes like Tert, DAZL, POUV, and NANOG. Cultured PGCs labeled with PKH26 were reinjected into Stage X recipient embryos and into the dorsal aorta of Stage 14-17 embryos to assay their ability of migration into the germinal crescent and gonads, respectively. In conclusion, the most suitable culture system for PGCs is as follows: feeder layer cells treated with 20 μg/mL mitomycin C for 2 hours, and with 50% conditioned medium added to the factor culture medium. PGCs cultured in this system retain their pluripotency and the unique ability of migration without transformation, indicating the successful preliminary establishment of chicken primordial germ cell lines and these PGCs can be considered for use as carriers in transgenic bioreactors. |
| first_indexed | 2024-04-13T12:50:45Z |
| format | Article |
| id | doaj.art-fe204c8f8b3e47248176d496181ca84a |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T12:50:45Z |
| publishDate | 2018-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-fe204c8f8b3e47248176d496181ca84a2022-12-22T02:46:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-01134e019645910.1371/journal.pone.0196459Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines.Linglin KongLingling QiuQixin GuoYing ChenXin ZhangBowen ChenYang ZhangGuobin ChangPrimordial germ cells (PGCs) are precursors of functional gametes and can be used as efficient transgenic tools and carriers in bioreactors. Few methods for long-term culture of PGCs are available. In this study, we tested various culture conditions for PGCs, and used the optimum culture system to culture chicken gonad PGCs for about three hundred days. Long-term-cultured PGCs were detected and characterized by karyotype analysis, immunocytochemical staining of SSEA-1, c-kit, Sox2, cDAZL, and quantitative RT-PCR for specific genes like Tert, DAZL, POUV, and NANOG. Cultured PGCs labeled with PKH26 were reinjected into Stage X recipient embryos and into the dorsal aorta of Stage 14-17 embryos to assay their ability of migration into the germinal crescent and gonads, respectively. In conclusion, the most suitable culture system for PGCs is as follows: feeder layer cells treated with 20 μg/mL mitomycin C for 2 hours, and with 50% conditioned medium added to the factor culture medium. PGCs cultured in this system retain their pluripotency and the unique ability of migration without transformation, indicating the successful preliminary establishment of chicken primordial germ cell lines and these PGCs can be considered for use as carriers in transgenic bioreactors.http://europepmc.org/articles/PMC5927411?pdf=render |
| spellingShingle | Linglin Kong Lingling Qiu Qixin Guo Ying Chen Xin Zhang Bowen Chen Yang Zhang Guobin Chang Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines. PLoS ONE |
| title | Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines. |
| title_full | Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines. |
| title_fullStr | Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines. |
| title_full_unstemmed | Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines. |
| title_short | Long-term in vitro culture and preliminary establishment of chicken primordial germ cell lines. |
| title_sort | long term in vitro culture and preliminary establishment of chicken primordial germ cell lines |
| url | http://europepmc.org/articles/PMC5927411?pdf=render |
| work_keys_str_mv | AT linglinkong longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT linglingqiu longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT qixinguo longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT yingchen longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT xinzhang longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT bowenchen longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT yangzhang longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines AT guobinchang longterminvitrocultureandpreliminaryestablishmentofchickenprimordialgermcelllines |