Background: The process of the assisted reproductive technology (ART) cycle is extremely complicated, and various factors in each step may influence the final clinical outcomes; thus, optimizing culture conditions for embryos is crucial in the ART cycle, particularly when the traditional petri-dish method remains unchanged for decades. In the current study, we intend to culture embryos in a dynamic environment on chips to optimize the embryo culture conditions.Methods: Multilayer soft lithography technology was utilized to establish a microfluidics-based oviduct. Mouse primary oviduct epithelial cells were identified by immunofluorescence staining and then loaded into the chip to coculture with the embryos. The development potential parameters of embryos on chips with cells, on chips without cells, and in drops were compared, as well as reactive oxygen species (ROS) in embryos.Results: There were no obvious differences regarding the fertilization rate, 4-Cell embryo rate, cleavage rate, high-quality embryo rate, or blastocyst formation rate. However, the intracellular ROS levels in 4-Cell stage embryos on chips with cells were statistically significantly lower than those in drops (P < 0.001). This organ-onchip device allowed the probability of mammalian embryo culture in a microfluidic-based manner.Conclusions: Our findings demonstrated that this novel oviduct-on-chip model may optimize embryo culture conditions by reducing intracellular ROS levels, which may be a competent alternative to the existing stable embryo culture system.