UC-MSCs appear to be a viable option for stem cell-based treatment. Cynomolgus monkeys, with their psychological and genetic parallels to humans, are essential models for medical study and development. Hypoxia is a basic characteristic of angiogenesis, glucose metabolism, and cell proliferation and survival in both healthy and pathological individuals. This study aimed to develop and characterize UC-MSCs requirements derived from the cynomolgus monkey. We have successfully developed UC-MSCs derived from Mf under hypoxic conditions. Mf UC-MSCs was cultured from the WJ area. Our Mf UC-MSCs have developed into adipocytes, osteocytes, and chondrocytes. The protein of the biomarker MSC was expressed in our hypoxic precondition Mf UC-MSCs and the mRNA expression levels of ERK1/2, AKT1, and HIF-1 alpha increased in the hypoxic condition compared to the normoxic and control groups. The JNK and NFκB genes were decreased in the hypoxic group. Interestingly, our hypoxic Mf UC-MSCs suppressed IL-6 expression. These findings show that preconditioned Mf UC-MSCs produce anti-inflammatory properties, making them potential candidates for regenerative medicine. Their response to hypoxia is crucial for developing targeted therapies to enhance regenerative capabilities in conditions such as ischemic injury and neurodegeneration. © 2026 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.