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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
摘要

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.

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Sigma-Aldrich
二甲基亚砜, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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Accutase® 溶液, sterile-filtered, suitable for cell culture
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纤连蛋白牛血浆, solution, sterile-filtered, BioReagent, suitable for cell culture
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TWEEN ® 20, for molecular biology, viscous liquid
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明胶 来源于猪皮肤, gel strength 300, Type A
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牛血清白蛋白 来源于牛血清, lyophilized powder, low endotoxin, BioReagent, suitable for cell culture, ≥98% (agarose gel electrophoresis)
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牛血清白蛋白 来源于牛血清, powder, BioXtra
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ESGRO®重组小鼠LIF蛋白, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^6 units/ml