Liu Xiaodan, Fan Qiuling, Yang Gang, Liu Nan, Chen Dong, Jiang Yi, Wang Lining
Department of Nephrology, The First Affiliated Hospital, China Medical University, Shenyang, Liaoning 110001, P.R. China.
Exp Ther Med. 2013 May;5(5):1322-1326. doi: 10.3892/etm.2013.1000. Epub 2013 Mar 12.
A modified procedure for Dynabead perfusion was developed to provide a practical methodology for obtaining large quantities of glomeruli from mice with a high purity. The glomeruli may be useful in exploring the mechanism behind glomerular diseases in conjunction with proteomics. The aim of the study was to save on costs and help researchers, particularly beginners, in the practical application of this method in their studies. Kidneys of C57BL/6 mice were perfused via two different techniques with Dynabeads. The purity and structures of the isolated glomeruli were investigated. The amounts of glomerular protein were measured and the costs of kidney and heart perfusions were compared. There was a 100% success rate at all stages involved in separating the glomeruli of mice via kidney perfusion. The isolated glomeruli remained intact and the purity was 96.7±1.2%. The average amounts of protein in the isolated glomeruli of 8- and 20-week-old mice were 45.6±13.4 and 55.8±17.0 g, respectively. The cost of glomerular isolation via kidney perfusion was one-fortieth of the cost of isolation via heart perfusion. The described procedure is practical and has a high success rate. The isolated glomeruli of mice were intact and pure and a large quantity was obtained at a lower cost.
开发了一种改良的磁珠灌注程序，以提供一种实用的方法，从小鼠身上获得大量高纯度的肾小球。这些肾小球在结合蛋白质组学探索肾小球疾病背后的机制方面可能会有用。该研究的目的是节省成本，并帮助研究人员，特别是初学者，在他们的研究中实际应用这种方法。通过两种不同的技术用磁珠对C57BL/6小鼠的肾脏进行灌注。研究了分离出的肾小球的纯度和结构。测量了肾小球蛋白的量，并比较了肾脏和心脏灌注的成本。通过肾脏灌注分离小鼠肾小球所涉及的所有阶段成功率均为100%。分离出的肾小球保持完整，纯度为96.7±1.2%。8周龄和20周龄小鼠分离出的肾小球中蛋白质的平均量分别为45.6±13.4和55.8±17.0μg。通过肾脏灌注分离肾小球的成本是通过心脏灌注分离成本的四十分之一。所描述的程序是实用的，成功率高。分离出的小鼠肾小球完整、纯净，且以较低成本获得了大量肾小球。
