Revealing cell populations catching the early stages of human embryo development in naive pluripotent stem cell cultures

• Published in: Stem cell reports 2023-01, Vol.18 (1), p.64-80
• Main Authors: Moya-Jódar, Marta, Ullate-Agote, Asier, Barlabé, Paula, Rodríguez-Madoz, Juan Roberto, Abizanda, Gloria, Barreda, Carolina, Carvajal-Vergara, Xonia, Vilas-Zornoza, Amaia, Romero, Juan Pablo, Garate, Leire, Agirre, Xabier, Coppiello, Giulia, Prósper, Felipe, Aranguren, Xabier L.
• Format: Article
• Language: English
• Subjects: 8CLCs; Blastocyst; Cell Differentiation - genetics; Embryo, Mammalian; Embryonic Development - genetics; Endoderm; Germ Layers; Humans; naive state; Pluripotent Stem Cells; single-cell RNA-seq; trophectoderm
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2022.11.015

Naive human pluripotent stem cells (hPSCs) are defined as the in vitro counterpart of the human preimplantation embryo’s epiblast and are used as a model system to study developmental processes. In this study, we report the discovery and characterization of distinct cell populations coexisting with epiblast-like cells in 5iLAF naive human induced PSC (hiPSC) cultures. It is noteworthy that these populations closely resemble different cell types of the human embryo at early developmental stages. While epiblast-like cells represent the main cell population, interestingly we detect a cell population with gene and transposable element expression profile closely resembling the totipotent eight-cell (8C)-stage human embryo, and three cell populations analogous to trophectoderm cells at different stages of their maturation process: transition, early, and mature stages. Moreover, we reveal the presence of cells resembling primitive endoderm. Thus, 5iLAF naive hiPSC cultures provide an excellent opportunity to model the earliest events of human embryogenesis, from the 8C stage to the peri-implantation period. [Display omitted] •Human naive cells cultured under 5iLAF condition show remarkable cellular heterogeneity•8C-like cells, characterized by concomitant expression of ZSCAN4 and MLT2A1 are present•Primitive endoderm-like cells and trophectoderm-like cells at different maturation stages are present•A population characterized by cellular stress and active LINE1 is present Moya-Jódar and colleagues address the void of knowledge on the heterogeneity present in hPSC naive cultures by the use of the single-cell RNA-seq technology. While epiblast-like cells represent the main cell population of naive 5iLAF culture, cells resembling the totipotent eight-cell stage human embryo, the trophectoderm, and the primitive endoderm cells of the blastocyst are also present. Thus, 5iLAF naive hiPSC cultures provide an excellent opportunity to model the earliest events of human embryogenesis.