ORIGINAL RESEARCH article
Front. Cell Dev. Biol.
Sec. Stem Cell Research
Volume 13 - 2025 | doi: 10.3389/fcell.2025.1566670
Optimal Sca-1-Based Procedure for Purifying Mouse Adipose-Derived Mesenchymal Stem Cells with Enhanced Proliferative and Differentiation Potential
Provisionally accepted
- 1Chongqing Emergency Medical Center, Chongqing, China
- 2Chongqing Emergency Medical Center, Chongqing University Central Hospital, School of Medicine, Chongqing University, Chongqing 400010, China, Chongqing, China
- 3Chongqing Emergency Medical Center, Chongqing University Central Hospital, School of Medicine, Chongqing University, Chongqing, China
- 4Department of Oncology, The First Affiliated Hospital of Chongqing Medical University, No.1 Youyi Road, Yuzhong District, Chongqing 400016, China, Chongqing, China
- 5Department of Intensive Care Medicine, Chongqing Emergency Medical Center, Chongqing University Central Hospital, Chongqing University, Jiankang Road No.1, YuZhong District, Chongqing 400016, China., Chongqing, China
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Adipose-derived mesenchymal stem cells (ADSCs) are promising candidates for mesenchymal stem cell (MSC) therapy due to their ease of isolation from the stromal vascular fraction (SVF) of adipose tissue. However, traditional isolation methods often result in mouse ADSCs with low purity and significant heterogeneity contributing to inconsistencies in results from preclinical and clinical studies. This is partly attributed to the lack of consensus on their surface markers. This study compared three purification methods for isolating mouse ADSCs based on Sca-1 positivity-direct adherence (ADSC-A), magnetic cell sorting followed by adherence (ADSC-M), and adherence to the third generation followed by magnetic cell sorting (ADSC-AM). Third-generation ADSCs were evaluated for proliferative activity, differentiation potential, and functional enrichment using proliferation assays, trilineage differentiation assays, and RNA sequencing. Flow cytometry was employed to assess Sca-1 positivity and the expression of positive (CD44, CD90, CD29) and negative markers (CD31, CD45) in the fourth-generation ADSCs. Among the three methods, ADSC-AM exhibited superior properties, including uniform morphology, enhanced proliferation, and over 95% expression of Sca-1 and CD29. While all methods supported trilineage differentiation, ADSC-AM demonstrated enhanced adipogenesis. Furthermore, RNA sequencing and pathway enrichment analysis revealed that ADSC-AM possessed unique potential in angiogenesis and immune regulation. These findings suggest that the ADSC-AM method offers a simple and reproducible approach for obtaining high-purity mouse ADSCs with better functional properties and provide a fundamental reference for understanding mouse ADSCs surface marker profiles.
Keywords: Adipose-derived mesenchymal stem cells, Sca-1, vascular stem cells, heterogeneity, purification, Magnetic activated cell sorting
Received: 25 Jan 2025; Accepted: 23 Apr 2025.
Copyright: © 2025 Tao, Wang, Yan, Cheng, Liu, Du and Niu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Huimin Du, Department of Oncology, The First Affiliated Hospital of Chongqing Medical University, No.1 Youyi Road, Yuzhong District, Chongqing 400016, China, Chongqing, China
Bailin Niu, Department of Intensive Care Medicine, Chongqing Emergency Medical Center, Chongqing University Central Hospital, Chongqing University, Jiankang Road No.1, YuZhong District, Chongqing 400016, China., Chongqing, China
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