Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
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In vitro culture of chicken embryonic stem cell-like cells

  • Bo Ram Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Hyeon Yang (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration)
  • Received : 2023.02.20
  • Accepted : 2023.03.08
  • Published : 2023.03.31
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Abstract

Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.

Keywords

Acknowledgement

This work was carried out with the support of "Animal Science & Technology Development (Project No. PJ01481702)" from National Institute of Animal Science (NIAS), Rural Development Administration (RDA), Korea.

References

  1. Eyal-Giladi H and Kochav S. 1976. From cleavage to primitive streak formation: a complementary normal table and a new look at the first stages of the development of the chick. I. General morphology. Dev. Biol. 49:321-337. https://doi.org/10.1016/0012-1606(76)90178-0
  2. Fuet A, Montillet G, Jean C, Aubel P, Kress C, Rival-Gervier S, Pain B. 2018. NANOG is required for the long-term establishment of avian somatic reprogrammed cells. Stem Cell Reports 11:1272-1286. https://doi.org/10.1016/j.stemcr.2018.09.005
  3. Lee BR, Kim H, Park TS, Moon S, Cho S, Park T, Lim JM, Han JY. 2007. A set of stage-specific gene transcripts identified in EK stage X and HH stage 3 chick embryos. BMC Dev. Biol. 7:60.
  4. Lee BR, Rengaraj D, Choi HJ, Han JY. 2020. A novel F-box domain containing cyclin F like gene is required for maintaining the genome stability and survival of chicken primordial germ cells. FASEB J. 34:1001-1017. https://doi.org/10.1096/fj.201901294r
  5. Lee BR, Yang H, Byun SJ, Park TS. 2023. Research Note: development of a chicken experimental model platform for induced pluripotent stem cells by using CRISPR/Cas9-mediated NANOG knock-in reporter DF1 cells. Poult. Sci. 102: 102425.
  6. Livak KJ and Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402-408. https://doi.org/10.1006/meth.2001.1262
  7. Pain B, Clark ME, Shen M, Nakazawa H, Sakurai M, Samarut J, Etches RJ. 1996. Long-term in vitro culture and characterisation of avian embryonic stem cells with multiple morphogenetic potentialities. Development 122:2339-2348. https://doi.org/10.1242/dev.122.8.2339
  8. Park TS, Hong YH, Kwon SC, Lim JM, Han JY. 2003. Birth of germline chimeras by transfer of chicken embryonic germ (EG) cells into recipient embryos. Mol. Reprod. Dev. 65:389-395. https://doi.org/10.1002/mrd.10304
  9. Perry MM. 1987. Nuclear events from fertilisation to the early cleavage stages in the domestic fowl (Gallus domesticus). J. Anat. 153:267.
  10. Petitte JN, Clark ME, Liu G, Verrinder Gibbins AM, Etches RJ. 1990. Production of somatic and germline chimeras in the chicken by transfer of early blastodermal cells. Development 108:185-189. https://doi.org/10.1242/dev.108.1.185
  11. Smith AG. 1992. Mouse embryo stem cells: their identification, propagation and manipulation. Semin. Cell Biol. 3:385-399. https://doi.org/10.1016/1043-4682(92)90010-S
  12. Su Y, Zhu J, Salman S, Tang Y. 2020. Induced pluripotent stem cells from farm animals. J. Anim. Sci. 98:skaa343.
  13. Watanabe M, Kinutani M, Naito M, Ochi O, Takashima Y. 1992. Distribution analysis of transferred donor cells in avian blastodermal chimeras. Development 114:331-338. https://doi.org/10.1242/dev.114.2.331