原代肾细胞。
Primary kidney cells.
作者信息
Taub M
机构信息
Biochemistry Department, State University of New York at Buffalo, USA.
出版信息
Methods Mol Biol. 1997;75:153-61. doi: 10.1385/0-89603-441-0:153.
Primary rabbit kidney epithelial cell cultures can be obtained that express renal proximal tubule functions. Toward these ends, renal proximal tubules are purified from the rabbit kidney by the method of Brendel and Meezan. To summarize, each kidney is perfused with iron oxide, which becomes associated with glomeruli. The renal cortex is sliced and homogenized to liberate nephron segments. Renal proximal tubules and glomeruli are purified by sieving. The glomeruli, covered with iron oxide, are removed using a magnet. After a brief collagenase treatment (to disrupt basement membrane), the tubules are plated in hormonally defined serum-free medium supplemented with 5 μg/mL bovine insulin, 5 μg/mL human transferrin, and 5 × 10 hydrocortisone. After 5–6 d of incubation, confluent monolayers are obtained that possess multicellular domes, indicative of their capacity for transepithelial solute transport.
可以获得表达肾近端小管功能的原代兔肾上皮细胞培养物。为此，采用布伦德尔和米赞的方法从兔肾中纯化肾近端小管。简要概括如下：向每个肾脏灌注氧化铁，使其与肾小球结合。将肾皮质切片并匀浆以释放肾单位节段。通过筛分纯化肾近端小管和肾小球。使用磁铁去除覆盖有氧化铁的肾小球。经过短暂的胶原酶处理（以破坏基底膜）后，将小管接种于添加有5μg/mL牛胰岛素、5μg/mL人转铁蛋白和5×10⁻⁵氢化可的松的激素限定无血清培养基中。孵育5 - 6天后，获得具有多细胞穹顶的汇合单层细胞，这表明它们具有跨上皮溶质转运的能力。
Zotero 插件