Metabolic suppression during mesodermal differentiation of embryonic stem cells identified by single-cell...

Area of Science:

• Developmental Biology
• Cell Biology
• Stem Cell Research

Background:

• Vascular Endothelial Growth Factor Receptor 2 (Flk-1) is a key marker for mesodermal progenitors derived from embryonic stem cells (ESCs).
• Flk-1-positive (Flk-1(+)) cells differentiate into hemangioblasts and cardiovascular progenitors at early and late stages, respectively.
• Understanding metabolic regulation during differentiation is crucial for regenerative medicine and developmental studies.

Purpose of the Study:

• To investigate the metabolic characteristics of Flk-1(+) and Flk-1(-) cells during embryonic stem cell differentiation.
• To identify potential subgroups based on metabolic activity within these cell populations.
• To explore the relationship between metabolic status and key developmental gene expression.

Main Methods:

• Separation of Flk-1(+) and Flk-1(-) cells from spontaneously differentiating mouse ESC-derived embryoid bodies (EBs).
• Analysis of cell aggregate size and oxygen consumption rate (OCR).
• Single-cell comprehensive gene expression analysis, including Nanog and Pou5f1 expression.

Main Results:

• Late-stage Flk-1(+) cell aggregates exhibited smaller size and lower OCR compared to Flk-1(-) cells.
• Both Flk-1(+) and Flk-1(-) cells segregated into subgroups with high or low glucose metabolic activity.
• Metabolic suppression was observed in cells with intermediate expression of Nanog and Pou5f1.

Conclusions:

• Temporary metabolic suppression is an intrinsic characteristic of mesodermal differentiation.
• Metabolic regulation is dynamically controlled during ESC differentiation and linked to specific progenitor populations.
• Gene expression patterns, particularly Nanog and Pou5f1, correlate with metabolic states during differentiation.