Maser Robin L, Magenheimer Brenda S, Calvet James P
Department of Clinical Laboratory Sciences, University of Kansas Medical Center, Kansas City, KS, United States; Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, United States; Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, KS, United States.
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, United States; Jared Grantham Kidney Institute, University of Kansas Medical Center, Kansas City, KS, United States.
Methods Cell Biol. 2019;153:169-183. doi: 10.1016/bs.mcb.2019.04.018. Epub 2019 Jun 13.
Metanephric organ culture, or ex vivo embryonic kidney culture, was developed in the mid-twentieth century as a means to understand the development of the mammalian kidney and was used in early studies of polycystic kidney disease to explore mechanisms of renal cyst initiation by non-genetic factors. Following the identification of cystogenic genes, a resurgence of the use of metanephric organ culture occurred and has yielded insight into basic mechanisms of cystic dilation; facilitated identification of pathogenic pathways and potential therapeutic targets; and provided a system for evaluating therapeutic agents. This chapter provides detailed, step-by-step protocols with rationale and tips for the establishment, maintenance and treatment of metanephric organ cultures, and for performance of the most commonly employed secondary analyses of these cultures.
后肾器官培养,即离体胚胎肾培养,是在20世纪中叶发展起来的,作为一种了解哺乳动物肾脏发育的手段,并在早期多囊肾病研究中用于探索非遗传因素引发肾囊肿的机制。在确定致囊肿基因后,后肾器官培养的应用再度兴起,并在囊性扩张的基本机制方面取得了进展;有助于识别致病途径和潜在治疗靶点;并提供了一个评估治疗药物的系统。本章提供了详细的、循序渐进的方案,包括建立、维持和处理后肾器官培养物的基本原理和技巧,以及对这些培养物进行最常用的二次分析的操作方法。