Bước đầu thiết lập quy trình thu nhận exosome từ dịch nuôi cấy tế bào gốc trung mô từ mô mỡ hướng đến ứng dụng trong y học tái tạo

Exosomes, small extracellular vesicles, are emerging as a revolutionary frontier in regenerative medicine due to their remarkable ability to facilitate tissue repair, regeneration, and restoration of damaged physiological functions. Their therapeutic potential in treating complex diseases is immense, positioning them as a powerful tool in future medical applications. However, to harness their full clinical and research potential, the establishment of standardized, safe, and quality-controlled isolation protocols is paramount. This study addresses this critical need by meticulously developing and evaluating a robust protocol for the isolation and characterization of exosomes derived from adipose tissue-derived mesenchymal stem cells, thereby contributing to the advancement of exosome-based therapies.

Researchers and practitioners in regenerative medicine, stem cell biology, biotechnology, and related medical fields.

The burgeoning field of regenerative medicine recognizes exosomes as key therapeutic agents capable of supporting tissue repair and recovery. This study aimed to address the critical need for a standardized and safe protocol for isolating high-quality exosomes, crucial for both research and potential clinical applications in regenerative medicine. The methodology involved isolating and expanding mesenchymal stem cells from human adipose tissue. The culture medium from these cells was then processed to isolate exosomes using a series of centrifugation steps. Characterization of the isolated exosomes was rigorously performed according to International Society for Extracellular Vesicles (ISEV) standards, including morphological assessment via scanning electron microscopy (SEM) and dynamic light scattering (DLS) for size determination, and flow cytometry for identifying specific surface protein markers such as CD9, CD63, and CD81. Furthermore, protein concentration was quantified using the Bradford method, and safety was ensured by evaluating sterility and the absence of heavy metals like lead, cadmium, arsenic, and mercury. The results demonstrated the successful isolation of spherical exosomes with particle sizes ranging from 30-200 nm, consistent with ISEV guidelines. Flow cytometry confirmed the expression of characteristic exosomal proteins CD9 (70.82%), CD63 (70.77%), and CD81 (95.03%). The exosome product achieved a protein concentration of approximately 45.22 µg/ml and was found to be free of surveyed heavy metals. These findings indicate that the isolated exosomes meet stringent international standards for safety and quality. This established protocol provides a reliable and high-quality source of exosomes, particularly relevant for advanced research and future therapeutic applications in regenerative medicine, paving the way for preclinical studies and ultimately clinical trials for tissue regeneration and disease treatment in Vietnam.