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Single organoid RNA-sequencing reveals high organoid-to-organoid variability

Kristin Gehling, View ORCID ProfileSwati Parekh, Farina Schneider, Marcel Kirchner, View ORCID ProfileVangelis Kondylis, View ORCID ProfileChrysa Nikopoulou, View ORCID ProfilePeter Tessarz
doi: https://doi.org/10.1101/2021.11.22.469588
Kristin Gehling
1Max Planck Research Group “Chromatin and Ageing”, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany
2Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Straße 26, 50931 Cologne, Germany
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Swati Parekh
1Max Planck Research Group “Chromatin and Ageing”, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany
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Farina Schneider
3Institute for Pathology, University Hospital Cologne, Kerpener Str. 62, 50937 Cologne, Germany
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Marcel Kirchner
4FACS & Imaging Core Facility, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany
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Vangelis Kondylis
3Institute for Pathology, University Hospital Cologne, Kerpener Str. 62, 50937 Cologne, Germany
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Chrysa Nikopoulou
1Max Planck Research Group “Chromatin and Ageing”, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany
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  • For correspondence: cnikopoulou@age.mpg.de ptessarz@age.mpg.de
Peter Tessarz
1Max Planck Research Group “Chromatin and Ageing”, Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Str. 9b, 50931 Cologne, Germany
2Cologne Excellence Cluster on Cellular Stress Responses in Aging-associated Diseases (CECAD), Joseph-Stelzmann-Straße 26, 50931 Cologne, Germany
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  • ORCID record for Peter Tessarz
  • For correspondence: cnikopoulou@age.mpg.de ptessarz@age.mpg.de
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ABSTRACT

Over the last decades, organoids have been established from the majority of tissue resident stem and iPS cells. They hold great promise for our understanding of mammalian organ development, but also for the study of disease or even personalized medicine. In recent years, several reports hinted at intraculture organoid variability, but a systematic analysis of such a heterogeneity has not been performed before. Here, we used RNA-seq of individual organoids to address this question. Importantly, we find that batch-to-batch variation is very low, even when prepared by different researchers. On the other hand, there is organoid-to-organoid variability within a culture. Using differential gene expression, we did not identify specific pathways that drive this variability, pointing towards possible effects of the microenvironment within the culture condition. Taken together, our study provides a framework for organoid researchers to properly consider experimental design.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted November 22, 2021.
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Single organoid RNA-sequencing reveals high organoid-to-organoid variability
Kristin Gehling, Swati Parekh, Farina Schneider, Marcel Kirchner, Vangelis Kondylis, Chrysa Nikopoulou, Peter Tessarz
bioRxiv 2021.11.22.469588; doi: https://doi.org/10.1101/2021.11.22.469588
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Single organoid RNA-sequencing reveals high organoid-to-organoid variability
Kristin Gehling, Swati Parekh, Farina Schneider, Marcel Kirchner, Vangelis Kondylis, Chrysa Nikopoulou, Peter Tessarz
bioRxiv 2021.11.22.469588; doi: https://doi.org/10.1101/2021.11.22.469588

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