Embryonic Stem Cell Culture Conditions Support Distinct States Associated with Different Developmental Stages and Potency

• ESC derivation condition does not irreversibly affect functional potency
• ESCs cultured in 2i and KOSR resemble early stages of embryonic development
• ESCs cultured in 2i and KOSR have enhanced functional potency
• ESCs cultured in KOSR resemble primitive endoderm

SummaryEmbryonic stem cells (ESCs) are cell lines derived from the mammalian pre-implantation embryo. Here we assess the impact of derivation and culture conditions on both functional potency and ESC transcriptional identity. Individual ESCs cultured in either two small-molecule inhibitors (2i) or with knockout serum replacement (KOSR), but not serum, can generate high-level chimeras regardless of how these cells were derived. ESCs cultured in these conditions showed a transcriptional correlation with early pre-implantation embryos (E1.5–E3.5) and contributed to development from the 2-cell stage. Conversely, the transcriptome of serum-cultured ESCs correlated with later stages of development (E4.5), at which point embryonic cells are more restricted in their developmental potential. Thus, ESC culture systems are not equivalent, but support cell types that resemble distinct developmental stages. Cells derived in one condition can be reprogrammed to another developmental state merely by adaptation to another culture condition.

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