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Human pluripotent stem cells (hPSCs) exist in at least two distinct states of pluripotency: naïve and primed. While naïve hPSCs possess the unique ability to generate blastocyst-like structures, they are often genetically and epigenetically unstable, which compromises the quality and developmental potential of naïve hPSC-derived blastoids. This protocol presents an optimized human blastoid protocol through a transient resetting method that converts primed hPSCs into a naïve-like state, addressing the stability issues associated with long-term naïve hPSC maintenance. The approach is compatible with both feeder-free and feeder-based culture systems and demonstrates high efficiency in generating human blastoids directly from primed hPSCs. This advancement provides a more robust and reliable strategy for blastoid formation, circumventing the limitations of suboptimal naïve hPSC cultures.

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Generating Human Blastoids Following Transient Naïve Resetting of Primed Pluripotent Stem Cells

  • 实验方案

  • 发表日期：

    2025年

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  • Maria Carolina Zimara,
  • Carlos A. Pinzon-Arteaga,
  • Kun Liu,
  • Toshihiko Ezashi,
  • Seiya Oura,
  • Shijian Lyu,
  • Menaka Sanghvi,
  • Ye Yuan,
  • Jun Wu

摘要

Human pluripotent stem cells (hPSCs) exist in at least two distinct states of pluripotency: naïve and primed. While naïve hPSCs possess the unique ability to generate blastocyst-like structures, they are often genetically and epigenetically unstable, which compromises the quality and developmental potential of naïve hPSC-derived blastoids. This protocol presents an optimized human blastoid protocol through a transient resetting method that converts primed hPSCs into a naïve-like state, addressing the stability issues associated with long-term naïve hPSC maintenance. The approach is compatible with both feeder-free and feeder-based culture systems and demonstrates high efficiency in generating human blastoids directly from primed hPSCs. This advancement provides a more robust and reliable strategy for blastoid formation, circumventing the limitations of suboptimal naïve hPSC cultures.