Butkowski R J, De A
Prep Biochem. 1982;12(3):209-27. doi: 10.1080/00327488208065563.
Methods were developed to obtain tubules and glomeruli and their respective basement membranes from mouse kidneys. While the procedures are especially useful for preparing tubules, one method can be used to simultaneously prepare both tubules and glomeruli. Tubules can be obtained from single animals, while a minimum of five animals are required in order to prepare glomeruli. Either minced whole kidneys or dissected cortex tissue is dispersed using a polytron and the desired fractions are obtained by sucrose density gradient centrifugation. discontinuous gradients consisting of 49.5, 53, 57, and 60% sucrose are used in the first method. Following centrifugation, tubules are collected on the 49.5% sucrose layer and glomeruli are pelleted on the bottom of the tube. In the second method, disrupted tissue is mixed with 57% sucrose and after centrifugation a layer of pure tubules is obtained from the top of the sucrose solution. Basement membranes are then obtained by a sonication method and the average yields per kidney are 0.1 mg and 0.02 mg for tubular and glomerular basement membranes respectively. Their chemical compositions are similar to the respective basement membranes from other species.
已开发出从小鼠肾脏获取肾小管、肾小球及其各自基底膜的方法。虽然这些程序对制备肾小管特别有用，但有一种方法可用于同时制备肾小管和肾小球。肾小管可从单只动物获得，而制备肾小球则至少需要五只动物。用组织捣碎机将切碎的全肾或解剖的皮质组织分散，通过蔗糖密度梯度离心获得所需组分。第一种方法使用由49.5%、53%、57%和60%蔗糖组成的不连续梯度。离心后，肾小管收集在49.5%蔗糖层上，肾小球沉淀在管底。在第二种方法中，将破碎的组织与57%蔗糖混合，离心后从蔗糖溶液顶部获得一层纯肾小管。然后通过超声处理方法获得基底膜，每只肾脏肾小管和肾小球基底膜的平均产量分别为0.1毫克和0.02毫克。它们的化学组成与其他物种各自的基底膜相似。
