Molecular strategies to enhance induced pluripotent stem cell reprogramming in buffalo (Bubalus bubalis)

Induced pluripotent stem cells (iPSCs) represent a valuable tool in regenerative biology and reproductive biotechnology due to their capacity for self-renewal and differentiation into all embryonic lineages. However, in buffalo ( Bubalus bubalis ), iPSC generation remains inefficient and insufficiently characterized when compared to model species. This review summarizes key molecular strategies aimed at improving reprogramming efficiency in this species. The role of canonical transcription factors (OCT4, SOX2, KLF4, and c-MYC) is well established; however, species-specific limitations restrict the direct application of protocols developed for other mammals. Epigenetic modulation has emerged as a critical approach, particularly through the use of histone deacetylase inhibitors (HDACis), which enhance chromatin accessibility, promote activation of endogenous pluripotency genes, and improve colony formation. Hypoxic culture conditions also play an important role by inducing metabolic reprogramming toward glycolysis, reducing oxidative stress, and facilitating epigenetic remodeling. In parallel, the WNT/β-catenin signaling pathway is a key regulatory axis in pluripotency, and its modulation has been associated with increased expression of pluripotency-related genes and improved stem cell maintenance. Overall, iPSC production in buffalo is influenced by species-specific molecular characteristics that require tailored optimization strategies. The integration of epigenetic, metabolic, and signaling pathway modulation represents a promising approach to enhance reprogramming efficiency and support advances in reproductive biotechnologies in this species.