Generation of induced pluripotent stem cells from Bornean orangutans

Bornean orangutans (Pongo pygmaeus) are an endangered non-human primate species. Induced pluripotent stem cells (iPSCs) offer a promising avenue for preserving genetic resources and studying evolutionary processes. In this study, we successfully generate Bornean orangutan iPSCs (o-iPSCs) from peripheral blood mononuclear cells using Sendai virus-mediated reprogramming. Furthermore, we transform primed o-iPSCs into a naïve pluripotent state using a novel 4i/L/A culture system. The resulting naïve o-iPSCs exhibit key features similar to human naïve stem cells, including upregulation of KLF17, DNMT3L, and DPPA3/5. We also observe significant activation of the WNT signaling pathway and X chromosome reactivation in the 4i/L/A o-iPSCs. Transcriptome analysis confirmed their resemblance to human naïve embryonic stem cells. Our findings contribute to the molecular understanding of naïve o-iPSCs and provide a foundation for orangutan conservation using advanced technologies. To enhance their utility for developmental and therapeutic studies, we subjected the primed o-iPSCs to a transformation process to induce a naïve pluripotent state using the 4i/L/A culture system with feeder cells.