Abstract：Induced pluripotent stem cells (iPSCs) possess both self-renewal capability and multidirectional differentiation potential. Compared with embryonic stem cells (ESCs), iPSCs are associated with fewer ethical concerns and exhibit broad application prospects in medical and agricultural fields. However, it is limited in its differentiation efficacy and clinical translation by the persistence of residual exogenous reprogramming genes. Although integration-free technologies for generating iPSCs in rodents and humans (Homo sapiens) have become increasingly mature, obtaining integration-free iPSCs lines with stable pluripotency traits in livestock species remains challenging. This review focused on the core strategies for producing integration-free livestock iPSCs (such as integration-free vector delivery, novel reprogramming factor combinations and culture systems). Key regulatory mechanisms underpinning successful cases in large animals like pigs (Sus scrofa domesticus) were specifically analyzed. It further evaluated their translational potential and challenges from the dual perspectives of sustainable agricultural development and biomedical applications, thereby providing insights for advancing research and industrial applications of genome-safe livestock iPSCs.
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