Reprogramming epiblast stem cells into pre-implantation blastocyst cell-like cells

Area of Science:

• Developmental Biology
• Stem Cell Biology
• Synthetic Biology

Background:

• Synthetic embryo systems (SESs) offer ethical alternatives for studying early development.
• Epiblast stem cell (EPISC) reprogramming has generated blastocyst-like hemispheres (BCLH).

Purpose of the Study:

• To comprehensively analyze cell fate induction within the EPISC-derived BCLH SES using single-cell RNA sequencing.
• To investigate gene expression dynamics and regulatory networks governing cell differentiation in this system.

Main Methods:

• Single-cell RNA sequencing (scRNA-seq) at key time points.
• Analysis of MERVL reporter gene expression and RNA velocity.
• Bioinformatic analysis of gene-regulatory networks and cell-fate resemblance to in silico models.

Main Results:

• Broad induction of the 2C-like reporter MERVL was observed.
• scRNA-seq identified three major cell populations with gene expression profiles similar to epiblast, primitive endoderm, and trophectoderm.
• Key gene-regulatory networks, zygotic genome activation genes, and RNA splicing patterns were enriched in induced blastocyst-like cell fates.

Conclusions:

• The EPISC-derived BCLH SES successfully induces extraembryonic cell populations.
• This system provides a valuable model for studying cell potency and early developmental processes.
• The findings contribute to advancing developmental biology and biomedicine.