L-Proline Supplementation Drives Self-Renewing Mouse Embryonic Stem Cells to a Partially Primed Pluripotent State: The Early Primitive Ectoderm-Like Cell

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2022 Apr 29

PMID 35486235

Authors

Hannah J Glover

Rachel A Shparberg

Michael B Morris

Affiliations

Soon will be listed here.

Abstract

Mouse embryonic stem cells (mESCs) can be grown under a variety of culture conditions as discrete cell states along the pluripotency continuum, ranging from the least mature "ground state" to being "primed" to differentiate. Cells along this continuum are demarcated by differences in gene expression, X chromosome inactivation, ability to form chimeras and epigenetic marks. We have developed a protocol to differentiate "naïve" mESCs to a "partially primed" state by adding the amino acid L-proline to self-renewal medium. These cells express the primitive ectoderm markers Dnmt3b and Fgf5, and are thus called early primitive ectoderm-like (EPL) cells. In addition to changes in gene expression, these cells undergo a morphological change to flattened, dispersed colonies, have an increased proliferation rate, and a predisposition to neural fate. EPL cells can be used to study the cell states along the pluripotency continuum, peri-implantation embryogenesis, and as a starting point for efficient neuronal differentiation.

Citing Articles

Signalling pathway crosstalk stimulated by L-proline drives mouse embryonic stem cells to primitive-ectoderm-like cells.

Glover H, Holliday H, Shparberg R, Winkler D, Day M, Morris M Development. 2023; 150(20).

PMID: 37823343 PMC: 10652046. DOI: 10.1242/dev.201704.

Capturing Transitional Pluripotency through Proline Metabolism.

Minchiotti G, DAniello C, Fico A, De Cesare D, Patriarca E Cells. 2022; 11(14).

PMID: 35883568 PMC: 9323356. DOI: 10.3390/cells11142125.

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