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Updated: Jul 9, 2025

Generation of Mice Derived from Induced Pluripotent Stem Cells

Generation of Mice Derived from Induced Pluripotent Stem Cells

Published on: November 29, 2012

Generation of Mouse Primitive Endoderm Stem Cells.

Yasuhide Ohinata^1,2, Atsunori Saraya¹, Haruhiko Koseki^1,2

¹Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Inohana, Chuo ward, Chiba, Japan.

|November 29, 2023

Summary

Researchers developed a new method to create primitive endoderm stem cells (PrESCs). These PrESCs can develop into all primitive endoderm lineages, offering a powerful tool for studying early development and embryo reconstitution.

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Area of Science:

Developmental Biology
Stem Cell Biology
Genetics

Background:

Blastocysts contain three primary cell lineages: epiblast (Epi), trophoblast (TB), and primitive endoderm (PrE).
While embryonic stem cells (ESCs) and trophoblast stem cells (TSCs) are established, primitive endoderm stem cell (PrESC) lines with full developmental potential were lacking.
Existing extraembryonic endoderm cells (XENCs) derived from PrE show limited contribution to specific extraembryonic tissues in vivo.

Purpose of the Study:

To establish and characterize fully potent primitive endoderm stem cell (PrESC) lines.
To provide a reliable in vitro model for studying primitive endoderm (PrE) development and function.
To enable studies on embryo reconstitution using a comprehensive set of stem cell types.

Main Methods:

Keywords:

Blastocyst Blastocyst complementation Extraembryonic endoderm Primitive endoderm Primitive endoderm stem cells Yolk sac

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PrESCs were established and maintained using mouse embryonic fibroblast (MEF) feeder cells in a serum-free medium.
The culture medium was supplemented with fibroblast growth factor 4 (FGF4), heparin, CHIR99021, and platelet-derived growth factor-AA (PDGF-AA).
PrESC identity and differentiation potential were assessed through marker expression analysis and blastocyst injection experiments.

Main Results:

Established PrESC lines co-express markers of pluripotency and endoderm lineage commitment, resembling in vivo PrE.
Transplantation of PrESCs into blastocysts resulted in efficient contribution to visceral endoderm (VE), parietal endoderm (PE), and marginal zone endoderm (MZE).
PrESCs demonstrated gene expression profiles and differentiation potential similar to endogenous PrE.

Conclusions:

The developed protocol successfully establishes fully potent PrESC lines.
PrESCs serve as a valuable model for investigating primitive endoderm biology and early developmental mechanisms.
PrESCs are crucial for in vitro embryo reconstitution studies, complementing ESCs and TSCs.