Temporal Gene Expression and DNA Methylation during Embryonic Stem Cell Derivation.

ABSTRACT:

Objective

Dual inhibition of mitogen-activated protein kinase (MAPK) kinase (also known as MEK) and transforming growth factor β (TGFβ) type I receptors by PD0325901 and SB431542, known as R2i has been introduced as a highly efficient approach to the generation of mouse embryonic stem cells (ESC). In the present study, we investigated the molecular mechanisms underlying ESC derivation in the R2i condition.

Materials and methods

In this experimental study, zona-free whole E3.5 blastocysts were seeded on mouse embryonic fibroblast (MEF) feeder cells in both R2i and serum conventional media. The isolated inner cell mass (ICM), ESCs and the ICM-outgrowths were collected on days 3, 5 and 7 post-blastocyst culture for quantitative real timepolymerase chain reaction (qRT-PCR) analy

SUBMITTER: Samadian A 

PROVIDER: S-EPMC6004995 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature