Metastable pluripotent states in NOD-mouse-derived ESCs

Area of Science:

• Stem cell biology
• Developmental biology
• Genetics

Background:

• Embryonic stem cells (ESCs) originate from the inner cell mass (ICM) of blastocysts.
• Epiblast stem cells (EpiSCs) are derived from the postimplantation epiblast and have limited developmental potential.
• The nonobese diabetic (NOD) mouse strain was previously considered unsuitable for ESC derivation.

Purpose of the Study:

• To characterize pluripotent states in the NOD mouse strain.
• To investigate factors influencing stem cell pluripotency in different genetic backgrounds.
• To explore the stability of stem cell identities in vitro.

Main Methods:

• Derivation and culture of stem cells from NOD mice.
• Utilizing constitutive expression of Klf4 or c-Myc for stabilization.
• Employing small molecules as replacements for reprogramming factors.
• Assessing pluripotency state transitions upon manipulation of exogenous factors.

Main Results:

• NOD stem cells can be stabilized into ESCs and induced pluripotent stem cells (iPSCs) using Klf4, c-Myc, or small molecule mimetics.
• NOD ESCs and iPSCs exhibit metastable pluripotency, reverting to an EpiSC-like state when exogenous factors are removed.
• Reintroduction of factors restores ICM-like pluripotency, demonstrating dynamic state switching.
• Stem cells from distinct genetic backgrounds can adopt varied in vitro pluripotency states.

Conclusions:

• The genetic background of stem cells influences their in vitro pluripotency states.
• The stability of these pluripotency states is governed by endogenous genetic factors.
• Exogenous factors can modulate and control the plasticity of stem cell identities.
• This research provides insights into the regulation of pluripotency and its potential manipulation.