Multipotency of Adipose-Derived Stem Cells from Cynomolgus Monkeys

Background Adipose tissue is a rich source, and adipose-derived stem cells (ADSCs) are easy to isolate and expand and pose a low risk of tumorigenicity. ADSCs therefore represent a promising source for autologous transplantation, while avoiding graft rejection and ethical concerns. The cynomolgus monkey ( Macaca fascicularis ) is a nonhuman primate (NHP) that closely resembles humans in terms of its anatomical, physiological, and metabolic characteristics and shares high DNA sequence similarity. However, the use of cynomolgus monkey ADSCs has not been reported to date. In the present study, we aimed to isolate ADSCs and characterize their stem cell properties as a potential alternative for animal experimentation. Methods Adipose tissue samples were obtained from the subabdominal region of six cynomolgus monkeys. The growth and mesenchymal stem cell phenotypes of the isolated primary cells were characterized by their spindle-shaped morphology, alkaline phosphatase activity, and expression of key pluripotency/MSC markers with PCR and flow cytometry. The differentiation potential of cells was assessed through induction in several lineage-specific culture media. The induction of osteogenesis was examined via Alizarin red staining, adipogenesis via Oil Red O staining, and chondrogenesis via Alcian blue staining. Neural differentiation was examined by qRT‒PCR, immunostaining with specific antibodies and transcriptome analysis. High-performance liquid chromatography (HPLC) was used to detect dopamine levels. Results The cells isolated from adipose tissue samples from Cynomolgus monkeys expressed key pluripotency and mesenchymal stem cell markers. These cells can differentiate into osteoblasts, chondrocytes, adipocytes and neurons, indicating that they are multipotent mesenchymal stem cells, which are known as ADSCs. Furthermore, cynomolgus monkey ADSCs can differentiate into neural progenitor cells or mature neural cells, such as dopamine-producing cells. Transcriptome sequencing and pathway analyses revealed upregulation of neural-related genes and conserved activation of key signaling pathways during neural differentiation, closely paralleling patterns observed in human ADSCs. Conclusions We demonstrated for the first time that adipose-derived stem cells (ADSCs) from cynomolgus monkeys are fully functional MSCs that can differentiate into osteogenic, adipogenic, chondrogenic, and neural lineages. Comprehensive characterization using PCR, immunostaining, flow cytometry, and transcriptomic analysis confirmed classic mesenchymal stem cell properties and strong expression of core ADSC markers, closely matching human ADSCs. Therefore, our current study represents a significant step toward establishing a reliable cell source for nonhuman primate disease modeling, which is essential for preclinical testing before human trials.