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  • Generating primed pluripotent epiblast stem cells: A methodology chapter.

Generating primed pluripotent epiblast stem cells: A methodology chapter.

Current topics in developmental biology (2020-03-30)
Milan Samanta, Sundeep Kalantry
ABSTRACT

At least two distinct pluripotent states, referred to as naïve and primed, define the early mammalian embryo. In the mouse, the pluripotent epiblast cells in the pre/peri-implantation embryo are the source of naïve embryonic stem cells (ESCs). After the embryo implants, the epiblast lineage generates a restricted or primed population of stem cells, referred to as epiblast stem cells (EpiSCs). ESCs can be cultured in EpiSC media to generate epiblast-like cells (EpiLCs). The differentiation of naive ESCs into primed EpiLCs permits insights into the development and differentiation of the pluripotent epiblast lineage. This chapter describes the generation and characterization of EpiSCs as well as EpiLCs.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid
Sigma-Aldrich
Bovine Serum Albumin, powder, BioXtra
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Bovine Serum Albumin, lyophilized powder, low endotoxin, BioReagent, suitable for cell culture, ≥98% (agarose gel electrophoresis)
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Fibronectin bovine plasma, solution, sterile-filtered, BioReagent, suitable for cell culture
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Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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Gelatin from porcine skin, gel strength 300, Type A
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Accutase® solution, sterile-filtered, suitable for cell culture
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^6 units/ml