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

Nat Methods
Authors
Keywords
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.

Year of Publication
2018
Journal
Nat Methods
Volume
15
Issue
9
Pages
732-740
Date Published
2018 09
ISSN
1548-7105
DOI
10.1038/s41592-018-0104-1
PubMed ID
30127506
PubMed Central ID
PMC6127858
Links
Grant list
P01 GM099134 / GM / NIGMS NIH HHS / United States
R01 GM067945 / GM / NIGMS NIH HHS / United States
R01 HD058013 / HD / NICHD NIH HHS / United States