Glomeruli from mice were isolated as described [33 (link)]. Briefly, mice were sacrificed and perfused with Dynabead M-450 (00388551, Invitrogen). The kidneys were digested in collagenase IV (1 mg/ml, Invitrogen) and pressed through a 100-μm cell strainer (BD Falcon, Bedford, MA), and then glomeruli were gathered using a magnetic concentrator, and the remaining fluids were centrifuged for collecting the tubules.
For isolation of rat glomeruli [36 (link)], SD rats were sacrificed in a sterile environment. Kidneys were harvested, decapsulated, minced, and then smashed down with a plunger through three sieves sequentially (200-100-60 mesh opening sizes). After washing and centrifugation, the glomeruli were resuspended in RPMI1640 with 10% FBS medium and plated on noncoated six-well plates for treatment with lentivirus expressing B7-1 (Genechem, Shanghai, China).
For isolation of rat glomeruli [36 (link)], SD rats were sacrificed in a sterile environment. Kidneys were harvested, decapsulated, minced, and then smashed down with a plunger through three sieves sequentially (200-100-60 mesh opening sizes). After washing and centrifugation, the glomeruli were resuspended in RPMI1640 with 10% FBS medium and plated on noncoated six-well plates for treatment with lentivirus expressing B7-1 (Genechem, Shanghai, China).
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