The kidney glomerulus is essential for proper kidney function. Until recently, technical challenges associated with glomerular isolation and subsequent dissolution into single cells have limited the detailed characterization of cells in the glomerulus. Previous techniques of kidney dissociation result in low glomerular cell yield, which limits high-throughput analysis. The ability to efficiently purify glomeruli and digest the tissue into single cells is especially important for single-cell characterization methods. Here, we present a detailed and comprehensive technique for the extraction and preparation of mouse glomerular cells, with high yield and viability. The method includes direct renal perfusion of Dynabeads via the renal artery followed by kidney dissociation and isolation of glomeruli by magnet; these steps provide a high number and purity of isolated glomeruli, which are further dissociated into single cells. The balanced representation of podocytes, mesangial and endothelial cells in single-cell suspensions of mouse glomeruli, and the high cell viability observed, confirm the efficiency of our method. With some practice, the procedure can be done in <3 h (excluding equipment setup and data analysis). This protocol provides a valuable technique for advancing future single-cell-based studies of the glomerulus in health, injury and disease.

肾小球对于正常的肾功能至关重要。直到最近，与肾小球分离和随后溶解成单细胞相关的技术挑战限制了肾小球细胞的详细表征。以前的肾分离技术导致肾小球细胞产量低，这限制了高通量分析。有效纯化肾小球并将组织消化成单个细胞的能力对于单细胞表征方法尤为重要。在这里，我们提出了一种用于提取和制备小鼠肾小球细胞的详细而全面的技术，具有高产量和活力。该方法包括通过肾动脉对 Dynabeads 进行直接肾灌注，然后通过磁铁解离肾小球；这些步骤提供了大量和纯度的分离肾小球，进一步分离成单个细胞。小鼠肾小球单细胞悬浮液中足细胞、系膜细胞和内皮细胞的平衡表现，以及观察到的高细胞活力，证实了我们方法的有效性。通过一些练习，该过程可以在 <3 小时内完成（不包括设备设置和数据分析）。该协议为推进未来基于单细胞的肾小球在健康、损伤和疾病方面的研究提供了一种有价值的技术。该过程可以在 <3 小时内完成（不包括设备设置和数据分析）。该协议为推进未来基于单细胞的肾小球在健康、损伤和疾病方面的研究提供了一种有价值的技术。该过程可以在 <3 小时内完成（不包括设备设置和数据分析）。该协议为推进未来基于单细胞的肾小球在健康、损伤和疾病方面的研究提供了一种有价值的技术。