Efficient isolation of interstitial fibroblasts directly from mouse kidneys or indirectly after ex vivo expansion.
Efficient isolation of interstitial fibroblasts directly from mouse kidneys or indirectly after ex vivo expansion.
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DOI：
10.1016/j.xpro.2021.100826
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发表时间：
2021-12-17
期刊：
影响因子：
--
通讯作者：
Suzuki N
中科院分区：
文献类型：
--
作者：
Nakai T;Iwamura Y;Suzuki N
肾间质成纤维细胞负责产生红系生长因子EPO和血管紧张素加压素，以及肾脏纤维化，在这些过程中它们转化为肌成纤维细胞。因此，对成纤维细胞的分析可以阐明肾脏疾病的复杂机制。然而，这些细胞的脆弱性使得它们的分离很难进行体外分析和体外培养。我们已经克服了这些困难，通过适度分离小鼠肾脏，并在半固体介质中与其他肾脏细胞共培养成纤维细胞。有关本议定书的使用和执行的完整细节，请参阅和。一种基于细胞分选仪的小鼠肾间质成纤维细胞分离方法一种从肾片中分离间质成纤维细胞的体外扩增方法用该方法分离的细胞可用于培养和单细胞分析肾间质成纤维细胞除负责肾脏纤维化外，还负责产生红系生长因子EPO和血管紧张素加压素，在这些细胞中它们转化为肌成纤维细胞。因此，对成纤维细胞的分析可以阐明肾脏疾病的复杂机制。然而，这些细胞的脆弱性使得它们的分离很难进行体外分析和体外培养。我们已经克服了这些困难，通过适度分离小鼠肾脏，并在半固体介质中与其他肾脏细胞共培养成纤维细胞。
Renal interstitial fibroblasts are responsible for producing the erythroid growth factor Epo and the vasopressor renin in addition to kidney fibrosis, in which they are transformed into myofibroblasts. Therefore, analyses of fibroblasts may elucidate the complex mechanisms of kidney diseases. However, the fragility of these cells makes their isolation for in vitro analyses and ex vivo cultivation difficult. We have overcome these difficulties by mildly dissociating mouse kidneys and coculturing fibroblasts with other kidney cells in semisolid medium. For complete details on the use and execution of this protocol, please refer to and. A cell sorter-based protocol for isolation of renal interstitial fibroblasts from mice A protocol for ex vivo expansion of interstitial fibroblasts from kidney pieces Cells isolated with this protocol are available for culture and single-cell analyses Renal interstitial fibroblasts are responsible for producing the erythroid growth factor Epo and the vasopressor renin in addition to kidney fibrosis, in which they are transformed into myofibroblasts. Therefore, analyses of fibroblasts may elucidate the complex mechanisms of kidney diseases. However, the fragility of these cells makes their isolation for in vitro analyses and ex vivo cultivation difficult. We have overcome these difficulties by mildly dissociating mouse kidneys and coculturing fibroblasts with other kidney cells in semisolid medium.
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