Abstract
In vitro induction of primordial germ cell like-cells (PGCLCs) from pluripotent stem cells (PSCs) is a robust method that will contribute to understanding the fundamentals of cell fate decisions, animal breeding, and future reproductive medicine. Here, we introduce this system established in the rat model. We describe a stepwise protocol to induce epiblast-like cells and subsequent PGCLCs by forming spherical aggregates from rat PSCs. We also describe a protocol to mature these PGCLCs from specified/migratory to the gonadal stage by aggregation with female gonadal somatic cells.
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Acknowledgments
We thank Dr. Roopsha Sengupta for editing the manuscript. Flow cytometry data was obtained in the University of Tokyo (IMSUT) FACS Core laboratory. This work was supported by KAKENHI from the Japan Society for the Promotion of Science grants (18H02367) to M.H and T.K, (18H05548, 20H03167) to T.K, (18H05544) to T.K, (19K23711) to M.O, AMED (JP18bm0704022, JP22bm1123008) to T.K, Cooperative Study Program (21-147) of National Institute for Physiological Sciences (NIPS) to T.K, research grants for young scientists (2020) of NIPS to M.O, and grants from The Sumitomo Foundation (210348) to T.K, Kato Memorial Bioscience foundation to T.K., and Daiichi Sankyo Foundation of Life Science to T.K.
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Oikawa, M., Hirabayashi, M., Kobayashi, T. (2024). Induction of Primordial Germ Cell-Like Cells from Rat Pluripotent Stem Cells. In: Barchi, M., De Felici, M. (eds) Germ Cell Development. Methods in Molecular Biology, vol 2770. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3698-5_8
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DOI: https://doi.org/10.1007/978-1-0716-3698-5_8
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