Prof. Chongqing University Central Hospital Yuzhong District, Chongqing, China (People's Republic)
Abstract: Adipose-derived mesenchymal stem cells (ADSCs) hold great promise for regenerative medicine due to their accessibility and multipotent differentiation capabilities. However, the purification of high-purity ADSCs remains a major challenge, hindered by inconsistent surface marker profiles and contamination with non-target cells. The inconsistency in results from preclinical and clinical studies largely arises from experimental models, especially mice, with developing ADSC surface markers. In this study, we developed a novel Sca-1-based purification strategy—adherence to the third generation followed by magnetic cell sorting (ADSC-AM)—to enhance the yield, purity, and functionality of ADSCs. Our comparative analysis of three purification methods—direct adherence (ADSC-A), magnetic cell sorting followed by adherence (ADSC-M), and ADSC-AM—demonstrated that the ADSC-AM method consistently outperformed the others. ADSC-AM cells exhibited a uniform spindle morphology, enhanced proliferative capacity, and superior adipogenic differentiation potential. Additionally, ADSC-AM achieved over 95% expression of Sca-1 and CD29, a critical improvement in purity. RNA sequencing and pathway enrichment analysis further revealed that ADSC-AM cells uniquely exhibited increased potential for osteogenic differentiation, angiogenesis, and immune regulation, highlighting their enhanced functional properties. This optimized Sca-1-based procedure represents an innovative and efficient approach for isolating ADSCs with superior purity and functional properties, providing a solid foundation for advancing research in stem cell-based therapies.
Funding Source: Support by the Fundamental Research Funds for the Central Universities [2022CDJYGRH-014], and Chongqing Medical Scientific Research Project [2023ZDXM012].
