肾脏的全器官分子分析可能遗漏了与肾小球疾病发病机理有关的重要因素。因此，需要通过分离富集的肾小球种群的技术来增强整个器官的分析。在这里，我们描述了如何使用差分法从新鲜组织中分离出大鼠肾小球悬浮液。其次,我们还展示了如何将它们用于原代肾小球系膜细胞培养物的繁殖。这些方案提供了用于下游分析的蛋白质和RNA分离的实用方法。这些技术容易适用于实验动物模型和人肾组织中分离的肾小球的研究。
Whole organ molecular analysis of the kidney potentially misses important factors involved in the pathogenesis of the glomerular disease. Organ-wide analysis therefore needs to be augmented by techniques that isolate enriched populations of glomeruli. Herein, we describe how differential sieving can be used to isolate a suspension of rat glomeruli from fresh tissue. Secondly, we also show how these can be used for the propagation of primary mesangial cell cultures. These protocols provide a practical approach for protein and RNA isolation for downstream analysis. These techniques are readily applicable to studies in isolated glomeruli in both experimental animal models and human kidney tissue.
