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  • Reduced MEK inhibition preserves genomic stability in naive human embryonic stem cells.

Reduced MEK inhibition preserves genomic stability in naive human embryonic stem cells.

Nature methods (2018-08-22)
Bruno Di Stefano, Mai Ueda, Shan Sabri, Justin Brumbaugh, Aaron J Huebner, Anna Sahakyan, Kendell Clement, Katie J Clowers, Alison R Erickson, Keiko Shioda, Steven P Gygi, Hongcang Gu, Toshi Shioda, Alexander Meissner, Yasuhiro Takashima, Kathrin Plath, Konrad Hochedlinger
ABSTRACT

Human embryonic stem cells (hESCs) can be captured in a primed state in which they resemble the postimplantation epiblast, or in a naive state where they resemble the preimplantation epiblast. Naive-cell-specific culture conditions allow the study of preimplantation development ex vivo but reportedly lead to chromosomal abnormalities, which compromises their utility in research and potential therapeutic applications. Although MEK inhibition is essential for the naive state, here we show that reduced MEK inhibition facilitated the establishment and maintenance of naive hESCs that retained naive-cell-specific features, including global DNA hypomethylation, HERVK expression, and two active X chromosomes. We further show that hESCs cultured under these modified conditions proliferated more rapidly; accrued fewer chromosomal abnormalities; and displayed changes in the phosphorylation levels of MAPK components, regulators of DNA damage/repair, and cell cycle. We thus provide a simple modification to current methods that can enable robust growth and reduced genomic instability in naive hESCs.

MATERIALS
Product Number
Brand
Product Description

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Anti-KLF17 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
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Mouse Anti-Stella Antibody, clone 3H5.2, Chemicon®, from mouse
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Triton X-100, laboratory grade