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Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level.
Reimer, Katharina C; Jansen, Jitske; Overheul, Gijs J; Miesen, Pascal; van Rij, Ronald P; Triana, Sergio H; Smeets, Bart; Schneider, Rebekka K; Kramann, Rafael.
  • Reimer KC; Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, Aachen, Germany; Institute of Cell and Tumor Biology, RWTH Aachen University, Aachen, Germany; Division of Nephrology and Clinical Immunology, RWTH Aachen University, Aachen, Germany. Electronic address: katharina.reimer
  • Jansen J; Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, Aachen, Germany; Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands; Department of Pediatric Nephrology, Radboud Institute for Molecular L
  • Overheul GJ; Department of Medical Microbiology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands.
  • Miesen P; Department of Medical Microbiology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands.
  • van Rij RP; Department of Medical Microbiology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, Nijmegen, the Netherlands.
  • Triana SH; Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany; Broad Institute of MIT and Harvard, Cambridge, MA, USA; Institute for Medical Engineering and Science, MIT, Cambridge, MA, USA; Department of Chemistry, MIT, Cambridge, MA, USA; Ragon Institute of
  • Smeets B; Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands.
  • Schneider RK; Institute of Cell and Tumor Biology, RWTH Aachen University, Aachen, Germany; Department of Developmental Biology, Erasmus Medisch Centrum, Rotterdam, the Netherlands; Oncode Institute, Erasmus Medisch Centrum, Rotterdam, the Netherlands; Department of Internal Medicine, Nephrology and Transplantati
  • Kramann R; Institute of Experimental Medicine and Systems Biology, RWTH Aachen University, Aachen, Germany; Division of Nephrology and Clinical Immunology, RWTH Aachen University, Aachen, Germany; Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medisch Centrum, Rotterdam, the Netherlan
STAR Protoc ; 3(3): 101612, 2022 09 16.
Article in English | MEDLINE | ID: covidwho-1937317
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).
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Induced Pluripotent Stem Cells / COVID-19 Limits: Humans Language: English Journal: STAR Protoc Year: 2022 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Induced Pluripotent Stem Cells / COVID-19 Limits: Humans Language: English Journal: STAR Protoc Year: 2022 Document Type: Article