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Merck

Generating primed pluripotent epiblast stem cells: A methodology chapter.

Current topics in developmental biology (2020-03-30)
Milan Samanta, Sundeep Kalantry
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At least two distinct pluripotent states, referred to as naรฏve and primed, define the early mammalian embryo. In the mouse, the pluripotent epiblast cells in the pre/peri-implantation embryo are the source of naรฏve 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
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Sigma-Aldrich
TWEENยฎ 20, for molecular biology, viscous liquid
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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-Maxโ„ข, sterile-filtered, BioReagent, suitable for hybridoma, โ‰ฅ99.7%
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Gelatin from porcine skin, gel strength 300, Type A
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Accutaseยฎ solution, sterile-filtered, suitable for cell culture
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ESGROยฎ Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^6 units/ml