PT  - JOURNAL ARTICLE
AU  - Gehling, Kristin
AU  - Parekh, Swati
AU  - Schneider, Farina
AU  - Kirchner, Marcel
AU  - Kondylis, Vangelis
AU  - Nikopoulou, Chrysa
AU  - Tessarz, Peter
TI  - Single organoid RNA-sequencing reveals high organoid-to-organoid variability
AID  - 10.1101/2021.11.22.469588
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.11.22.469588
4099  - http://biorxiv.org/content/early/2021/11/22/2021.11.22.469588.short
4100  - http://biorxiv.org/content/early/2021/11/22/2021.11.22.469588.full
AB  - 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 StatementThe authors have declared no competing interest.