Objectives: In recent years, many laboratories have transitioned from using human Embryonic Stem Cells (hESC) to human Induced Pluripotent Stem Cells (hiPSC). This shift has been driven by the ability to utilize hiPSCs with specific genomic characteristics or to reduce administrative complexities. However, there's a growing demand for new hESC lines that align with current ethical standards, culture practices ensuring genomic stability, and permissions for full genomic sequencing and low-passage banking. Our objective is to address this need by generating and thoroughly characterizing new human Embryonic Stem Cell lines.

Methods We developed a protocol to derive hESC lines from surplus embryos under updated ethical and legal consents. Our derivation protocol includes defined culture conditions optimized for genomic integrity, low-passage cryopreservation, and systematic genome sequencing. We tested the pluripotency and developmental potential of the lines through differentiation into multiple organoid systems. In parallel, we are deriving naïve hESC and trophoblast stem cells (TSCs) from the same line, enabling modeling of both pre- and post-implantation stages.

Results Our efforts have yielded a novel fully sequenced hESC line, demonstrating broad differentiation potential into various organoid types. The associated consents enable their use in developing advanced stem cell-based embryo models. This well-characterized hESC line will be deposited in cell banks, making them readily accessible to the broader scientific community. The successful derivation of naive and TSC lines from this hESCs opens new avenues for investigating early human development.

Conclusion This new hESC line fills an important gap in the current stem cell landscape by combining extended legal usage rights with broad experimental applicability. It will be made available to the scientific community through cell banks and will serve as reference line for initiatives like the “Infrastructure Organoid de Nantes” (ION). Furthermore, this line provides a robust platform for human disease modeling via genome editing.

Key words: hESC, hTSC, hNESC, quality control, differentiation