Baz Mohammad-H, Valette Marion, André Mireille, Varin Audrey, Trevisiol Emmanuelle, Sengenès Coralie, Gue Anne-Marie
LAAS-CNRS, Université de Toulouse, 31031, Toulouse, France.
RESTORE Research Center, Université de Toulouse, CNRS, Inserm, EFS, Toulouse, France.
Sci Rep. 2025 Mar 26;15(1):10471. doi: 10.1038/s41598-025-94353-y.
Mesenchymal stromal cells (MSCs) hold significant promise for their therapeutic potential and their possible role as disease biomarkers. While evidence suggests the presence of circulating Adipose-derived MSC (ASC) in peripheral blood (PB), isolating them is particularly challenging due to their low abundance, size variability, and incomplete characterization of their native immunophenotype in PB. Consequently, the relationship between ASC frequency in blood and various physiological or pathological conditions has been underexplored. In this study, we introduce ASC-Finder, a label-free isolation method specifically designed for adipose stromal cells (ASCs), a key MSC population. ASC-Finder integrates two independent modules: a size-dependent hydrodynamic filtration unit for sorting erythrocytes directly from PB and a negative enrichment module based on immunological markers to deplete remaining leukocytes. The device enabled removal of 99.98% of erythrocytes while achieving high recovery rates of spiked ASCs (> 81%) at rare-event concentrations (< 100 ASC/mL blood). Remarkably, ASC-Finder operates without clogging, even after multiple runs with donor blood samples. Crucially, our method bypasses the need for harsh lysis, centrifugation, or dilution buffers, preserving both cell integrity and phenotype-key factors for the discovery of novel cellular events. This work represents a significant advancement in the direct enrichment of circulating ASCs from whole PB without cell lysis, offering a crucial step toward investigating the characterization and role of blood-circulating ASCs.
间充质基质细胞（MSCs）因其治疗潜力以及作为疾病生物标志物的可能作用而具有巨大的前景。虽然有证据表明外周血（PB）中存在循环脂肪来源的间充质基质细胞（ASC），但由于其丰度低、大小可变以及在PB中其天然免疫表型的不完全表征，分离它们极具挑战性。因此，血液中ASC频率与各种生理或病理状况之间的关系尚未得到充分探索。在本研究中，我们引入了ASC-Finder，这是一种专门为脂肪基质细胞（ASCs）设计的无标记分离方法，脂肪基质细胞是关键的间充质基质细胞群体。ASC-Finder整合了两个独立模块：一个基于大小的流体动力学过滤单元，用于直接从PB中分选红细胞；以及一个基于免疫标记的阴性富集模块，用于去除剩余的白细胞。该设备能够去除99.98%的红细胞，同时在罕见事件浓度（<100个ASC/mL血液）下实现高回收率的加标ASCs（>81%）。值得注意的是，即使在对供体血样进行多次运行后，ASC-Finder也能正常运行而不会堵塞。至关重要的是，我们的方法无需进行苛刻的裂解、离心或稀释缓冲液处理，从而保留了细胞完整性和表型——这是发现新细胞事件的关键因素。这项工作代表了在不进行细胞裂解的情况下直接从全PB中富集循环ASCs方面的重大进展，为研究血液循环ASCs的特征和作用迈出了关键一步。
