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

ABSTRACT: 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 organoid-derived 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).

SUBMITTER: Reimer KC

PROVIDER: S-EPMC9293950 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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

Reimer Katharina C KC Jansen Jitske J Overheul Gijs J GJ Miesen Pascal P van Rij Ronald P RP Triana Sergio H SH Smeets Bart B Schneider Rebekka K RK Kramann Rafael R

STAR protocols 20220719 3

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 organoid-derived 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 organo ...[more]

PMID: 35983169

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