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

Abstract

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.

The many faces of Pluripotency: in vitro adaptations of a continuum of in vivo states

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A new genetic tool to improve immune‐compromised mouse models: Derivation and CRISPR/Cas9‐mediated targeting of NRG embryonic stem cell lines

“…It has been previously shown that 2i‐supplemented defined culture media support clonogenicity more efficiently than serum‐containing media (Martin Gonzalez et al, ). This could explain the ability of 2i to promote the derivation of ES cells of genetic backgrounds that are otherwise refractory when cultured in standard conditions, such as the NRG strain.…”

Advances in stem cells and regenerative medicine: single-cell dynamics, new models and translational perspectives

“…Mouse cells with the capacity to give rise to both embryonic and extraembryonic tissues were also described by Hongkui Deng (Peking University, China) and termed extended pluripotent stem cells (EPS cells) (Yang et al, 2017). Previous work had reported cells with similar properties using different culture media (Guo et al, 2009;Martin Gonzalez et al, 2016), but this work interestingly shows that human EPS cells contribute to both embryonic and extraembryonic tissue when transplanted into mice, which is not the case for human ESCs. Delineating the mechanisms behind the development of these highly plastic cells will provide important insight into the molecular underpinnings of stem cell reprogramming ability.…”