Generation of chicken induced pluripotent stem cells (iPSCs) from somatic cells using multi-lentiviral transduction and OAC2

The chicken iPSCs have been successfully established and exhibit significant potential for the recovery and conservation of endangered avian species. However, the efficiency of induction remains quite limited. To enhance the induction system, a six-factor reprogramming approach incorporating OCT4, SOX2, NANOG, LIN28, KLF4, and C-MYC (OSNLKM) was employed via lentiviral infection, in combination with the small-molecule compound OAC2 and mouse embryonic fibroblast (MEF) feeder layers. The results demonstrated that the six-factor system significantly accelerated the reprogramming process and improved induction efficiency compared to the conventional four-factor system (OCT4, SOX2, NANOG, and LIN28; OSNL). Additionally, the MEF feeder layers were found to be essential for the successful reprogramming of chicken somatic cells. The inclusion of Oct4-activating compound 2 (OAC2) further optimized the reprogramming conditions, enabling the successful induction of iPSCs from chicken embryonic fibroblasts (CEFs) and Sertoli cells. Collectively, this study establishes an optimized reprogramming system for chicken somatic cells, enhancing iPSCs induction efficiency and expanding the potential applications of iPSCs technology in avian conservation and genetic modification.