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Generation of mouse-human chimeric embryos.

Boyang Zhang1, Hanqin Li1, Zhixing Hu1

  • 1Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, NY, USA.

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|July 3, 2021
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Summary

Researchers developed a protocol to convert primed human pluripotent stem cells (hPSCs) into naive hPSCs. These naive hPSCs can generate mouse-human chimeric embryos, enabling the study of human cell development in vivo.

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Area of Science:

  • Developmental Biology
  • Stem Cell Biology
  • Chimerism

Background:

  • Human pluripotent stem cells (hPSCs) are crucial for regenerative medicine.
  • Generating mouse-human chimeric embryos allows for in vivo study of human cell development.
  • Converting primed hPSCs to a naive state is essential for efficient chimerism.

Purpose of the Study:

  • To establish a protocol for converting primed hPSCs to naive hPSCs.
  • To generate mouse-human chimeric embryos using these converted naive hPSCs.
  • To evaluate the potential of this protocol for studying hPSC development and generating human tissues in vivo.

Main Methods:

  • Primed hPSCs treated with a mammalian target of rapamycin inhibitor (Torin1).
  • Cells cultured in 2iLI medium on mouse embryonic fibroblasts to establish naive hPSCs.
  • Naive hPSCs injected into mouse blastocysts to create chimeric embryos.
  • Human cell contribution quantified using next-generation sequencing of 18S ribosomal DNA amplicons.

Main Results:

  • Successful conversion of primed hPSCs to naive hPSCs with mouse embryonic stem cell morphology.
  • Generation of E17.5 mouse embryos containing 0.1-4.0% human cells.
  • Demonstrated suitability of the protocol for studying hPSC development in a mammalian environment.

Conclusions:

  • A robust protocol for generating naive hPSCs from primed cells is described.
  • This method enables the creation of mouse-human chimeric embryos with significant human cell contribution.
  • The protocol holds promise for advancing research in human cell development, tissue engineering, and organ generation in animals.