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Maintenance of the ES Cell State

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The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...

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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation

Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation

Published on: October 20, 2014

Embryonic stem cell potency fluctuates with endogenous retrovirus activity.

Todd S Macfarlan¹, Wesley D Gifford, Shawn Driscoll

¹Howard Hughes Medical Institute, Gene Expression Laboratory, The Salk Institute for Biological Studies, 10010 North Torrey Pines, La Jolla, California 92037, USA.

|June 23, 2012

Summary

Researchers discovered rare two-cell (2C)-like stem cells in mouse cultures. These cells, unlike typical pluripotent stem cells, can form both embryonic and extraembryonic tissues, offering new insights into early development and cell-fate regulation.

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Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells (iPSCs) Using Retroviral Vector with GFP

Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells (iPSCs) Using Retroviral Vector with GFP

Published on: April 3, 2012

Area of Science:

Developmental Biology
Stem Cell Biology
Genomics

Background:

Embryonic stem (ES) cells, derived from blastocysts, resemble the inner cell mass but cannot form all extraembryonic tissues.
Pluripotency in ES cells is typically associated with specific protein markers like Oct4, Sox2, and Nanog.

Purpose of the Study:

To identify and characterize a rare cell population in mouse ES and induced pluripotent stem (iPS) cell cultures with totipotent potential.
To investigate the molecular mechanisms and cell-fate regulation governing this transient totipotent-like state.

Main Methods:

Genetic tagging of rare cells within ES and iPS cultures.
Analysis of pluripotency protein expression (Oct4, Sox2, Nanog).
Assessment of contribution to embryonic and extraembryonic tissues.
Transcriptome sequencing and bioinformatic analysis of 2C-like cells.
Investigation of the role of histone-modifying enzymes.

Main Results:

A rare, transient cell population expressing two-cell (2C) embryo transcripts was identified in mouse ES and iPS cells.
These 2C-like cells lack key pluripotency markers (Oct4, Sox2, Nanog) and exhibit totipotency, contributing to both embryonic and extraembryonic lineages.
Nearly all ES cells were observed to cycle through this totipotent-like state, influenced by histone-modifying enzymes.
Bioinformatic analysis revealed that many 2C transcripts originate from endogenous retroviral long terminal repeats, suggesting their role in cell-fate regulation.

Conclusions:

A transient totipotent-like state exists within standard mouse ES and iPS cell cultures, distinct from the typical pluripotent state.
This 2C-like state, regulated by epigenetic factors and potentially endogenous retroviral elements, expands the developmental potential of stem cells.
Understanding this state provides novel insights into early mammalian development and cell-fate plasticity for regenerative medicine applications.