Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level

被引：1
作者：
Reimer, Katharina C. [1 ,2 ,3 ]
Jansen, Jitske [1 ,4 ,5 ]
Overheul, Gijs J. [6 ]
Miesen, Pascal [6 ]
van Rij, Ronald P. [6 ]
Triana, Sergio H. [7 ,8 ,9 ,10 ,11 ]
Smeets, Bart
Schneider, Rebekka K. [2 ,12 ,13 ,14 ]
Kramann, Rafael [1 ,3 ,14 ]
机构：
[1] Rhein Westfal TH Aachen, Inst Expt Med & Syst Biol, Aachen, Germany
[2] Rhein Westfal TH Aachen, Inst Cell & Tumor Biol, Aachen, Germany
[3] Rhein Westfal TH Aachen, Div Nephrol & Clin Immunol, Aachen, Germany
[4] Radboud Univ Nijmegen, Radboud Inst Mol Life Sci, Dept Pathol, Med Ctr, Nijmegen, Netherlands
[5] Radboud Univ Nijmegen, Amalia Childrens Hosp, Radboud Inst Mol Life Sci, Dept Pediat Nephrol,Med Ctr, Nijmegen, Netherlands
[6] Radboud Univ Nijmegen, Radboud Inst Mol Life Sci, Dept Med Microbiol, Med Ctr, Nijmegen, Netherlands
[7] European Mol Biol Lab, Struct & Computat Biol Unit, Heidelberg, Germany
[8] Broad Inst MIT & Harvard, Cambridge, MA USA
[9] MIT, Inst Med Engn & Sci, Cambridge, MA USA
[10] MIT, Dept Chem, Cambridge, MA USA
[11] Ragon Inst MGH & Harvard, Cambridge, MA USA
[12] Erasmus MC, Dept Dev Biol, Rotterdam, Netherlands
[13] Erasmus MC, Oncode Inst, Rotterdam, Netherlands
[14] Erasmus MC, Dept Internal Med Nephrol & Transplantat, Rotterdam, Netherlands
来源：
STAR PROTOCOLS | 2022年 / 3卷 / 03期
基金：
欧洲研究理事会;
关键词：
Cell biology; Microbiology; Organoids; Single cell; Stem cells;
D O I：
10.1016/j.xpro.2022.101612
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
We describe a protocol for single-cell RNA sequencing of SARS-CoV-2-infected human induced pluripotent stem cell (iPSC)-derived kidney organoids. After inoculation of kidney organoids with virus, we use mechanical and enzymatic disruption to obtain single cell suspensions. Next, we process the organoidderived cells into sequencing-ready SARS-CoV-2-targeted libraries. Subsequent sequencing analysis reveals changes in kidney cells after virus infection. The protocol was designed for kidney organoids cultured in a 6-well transwell format but can be adapted to organoids with different organ backgrounds.For complete details on the use and execution of this protocol, please refer to Jansen et al. (2022).
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页数：18
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