Sun Heng, Xu Xiaoling, Deng Chuxia
Faculty of Health Sciences, University of Macau, Macau SAR, China.
Bio Protoc. 2020 Feb 20;10(4):e3530. doi: 10.21769/BioProtoc.3530.
Single cell RNA sequencing is a very powerful means for cellular heterogeneous studies and so becoming wildly utilized nowadays. To guarantee the success of such analysis, it is very important, though sometimes difficult, to obtain single cells suspension with high quality, especially from the primary solid organs like mammary glands. Digestion of mouse or human mammary glands with enzymes was previously described. However, the yield, viability, especially the separation degree of the cells have rarely been noticed in these studies. Here we described a detailed protocol for the single epithelial cells suspension preparation from mouse mammary glands, which could be applied for single cell RNA sequencing on different platforms. This protocol could be well adapted for dissociation of other solid organs and tumors, and the single cell suspension could be also used for many other experiments.
单细胞RNA测序是细胞异质性研究的一种非常强大的手段，如今正被广泛应用。为确保此类分析的成功，获得高质量的单细胞悬液非常重要，尽管有时很困难，尤其是从乳腺等原发性实体器官中获取时。此前已有用酶消化小鼠或人乳腺的描述。然而，这些研究中很少关注细胞的产量、活力，尤其是细胞的分离程度。在这里，我们描述了一种从小鼠乳腺制备单个上皮细胞悬液的详细方案，该方案可应用于不同平台的单细胞RNA测序。该方案可很好地适用于其他实体器官和肿瘤的解离，所得单细胞悬液也可用于许多其他实验。
