Establishment of novel common marmoset embryonic stem cell lines under various conditions.

Abstract Pluripotent stem cells (PSCs), embryonic stem cells (ESCs), and induced PSCs (iPSCs) are excellent tools for studying embryonic development in organisms and classified into naive and primed states. ESC-derived germline chimera individuals can be produced by injecting naive ESCs/iPSCs into preimplantation embryos, and conversion of primed human ESCs/iPSCs into a naive state provides insights into epiblast cell features. Non-human ESCs/iPSCs are alternatives to human naive ESCs/iPSCs, which elicit ethical issues. In this study, we used the common marmoset (Callithrix jacchus) as an animal model. Since 1996, 16 marmoset ESC lines have been established. Because most of these ESC lines are female and were derived >10 years ago, new ESCs, particularly male marmoset ESC lines, are needed. Here, we successfully established 17 novel marmoset ESC lines, including six male ESC lines from in vitro-fertilized (IVF) embryos and 12 ESC lines under feeder-free conditions. This report is the first to establish ESC lines using feeder-free conditions and IVF preimplantation blastocysts in marmosets, and these novel ESC lines could potentially facilitate future non-human primate ESC studies.