Single-cell Analysis of ACE2 Expression in Human Kidneys and Bladders Reveals a Potential Route of 2019-nCoV Infection

Wei Lin; Longfei Hu; Yan Zhang; Joshua D. Ooi; Ting Meng; Peng Jin; Xiang Ding; Longkai Peng; Lei Song; Zhou Xiao; Xiang Ao; Xiangcheng Xiao; Qiaoling Zhou; Ping Xiao; Jue Fan; Yong Zhong

The fresh kidney tissue was immediately transferred into the GEXSCOPE Tissue Preservation Solution (Singleron Biotechnologies) at 2-8°C. The samples were digested in 2ml GEXSCOPE Tissue Dissociation Solution (Singleron Biotechnologies) at 37°C for 15min in a 15ml centrifuge tube with continuous agitation after washed with Hanks Balanced Salt Solution (HBSS) for three times and cut into approximately 1-2 mm pieces. Subsequently, cell debris and other impurities were filtered by a 40-micron sterile strainer (Corning). The cells were centrifuged at 1000 rpm and 4°C for 5 minutes. Cell pellets were resuspended into 1ml PBS (HyClone). To remove red blood cells, 2 mL GEXSCOPE Red Blood Cell Lysis Buffer (Singleron Biotechnologies) was added to the cell suspension and incubated at 25°C for 10 minutes. The mixture was then centrifuged at 1000 rpm for 5 min and the cell pellet resuspended in PBS. Cells were counted with TC20 automated cell counter (Bio-Rad).